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Showing items 6041 to 6060 of 7642
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Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 03 February 2017
- Publication where the measure is specified
- The Uganda Gazette, Vol. CX No. 7
- Regulation where the measure is specified
- US EAS 849:2016, Silk (sheen) emulsion paint for interior use — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies requirements, sampling and test methods for silk (sheen) emulsion paint for interior use.
- Reference of the measure
- US EAS 849:2016
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- silk (sheen) emulsion paint for interior use
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 03 February 2017
- Publication where the measure is specified
- The Uganda Gazette, Vol. CX No. 7
- Regulation where the measure is specified
- US EAS 848:2016, Water-thinned priming paints for wood —Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies requirements, sampling and test methods for water-thinned priming paints intended for application by brush, roller spray or any other suitable method to the exterior and interior of soft wood joinery.
- Reference of the measure
- US EAS 848:2016
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- water-thinned priming paints
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 12 April 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 17 of 12th April 2013
- Regulation where the measure is specified
- US 945-1:2012, Pre-insulated flexible pipe systems — Part.1: Classification, general requirements and methods of test
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies the methods of test for flexible, pre-insulated, directly buried district heating pipe systems. Depending on the pipe assembly, this standard can be used for maximum operating temperatures of 95 °C to 140 °C and operating pressures of 6 bar to 25 bar. The pipe systems are designed for a lifetime of 30 years. For pipe systems with plastic service pipes, the respective temperature profiles are defined in US 945-2.
- Reference of the measure
- US 945-1:2012
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- Pre-insulated flexible pipe systems
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 03 February 2017
- Publication where the measure is specified
- The Uganda Gazette, Vol. CX No. 7
- Regulation where the measure is specified
- US 1603: 2016, Chia seed — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies the requirements, sampling and test methods for chia seed (Salvia hispanica L.) for human consumption. This standard does not apply to chia seed as a planting material.
- Reference of the measure
- US 1603:2016
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- chia seed (Salvia hispanica L.)
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 25 October 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 54 of 25th October 2013
- Regulation where the measure is specified
- US 979:2013, Breakfast cereals — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies quality requirements, contaminants, hygiene, packaging, labelling, methods of sampling and test for breakfast cereals intended for human consumption.
- Reference of the measure
- US 979:2013
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- breakfast cereals
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 07 August 2015
- Publication where the measure is specified
- THE UGANDA GAZETTE Vol. CVIII No. 43 of 7 August 2015
- Regulation where the measure is specified
- US IEC 60502-4:2010, Power cables with extruded insulation and their accessories for rated voltages from 1 kV (Um = 1,2 kV) up to 30 kV (Um = 36 kV) - Part 4: Test requirements on accessories for cables with rated voltages from 6 kV (Um = 7,2 kV) up
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies the test requirements for type testing of accessories for power cables with rated voltages from 3,6/6 (7,2) kV up to 18/30 (36) kV, complying with IEC 60502-2. (This Uganda Standard cancels and replaces, US EAS 506-4:2008, Power cables with extruded insulation and their accessories for rated voltages from 1 kV (Um = 1.2 kV) up to 30 kV (Um = 36 kV) — Part 4: Test requirements on accessories for cables with rated voltages from 6 kV (Um = 7.2 kV) up to 30 kV (Um = 36 kV), which has been republished)
- Reference of the measure
- US IEC 60502-4:2010
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- Power cables
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 12 April 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 17 of 12th April 2013
- Regulation where the measure is specified
- US EAS 782:2012, Composite flour – Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies the methods of sampling for test and test methods for composite flour intended for human consumption.
- Reference of the measure
- US EAS 782:2012
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- composite flour
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 12 April 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 17 of 12th April 2013
- Regulation where the measure is specified
- US EAS 781:2012, Biscuits — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies quality requirements, food additives, contaminants, hygiene, packaging, labelling, methods of sampling and test for biscuits.
- Reference of the measure
- US EAS 781:2012
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- biscuits
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 12 April 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 17 of 12th April 2013
- Regulation where the measure is specified
- US EAS 780:2012, Fresh cassava leaves — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies quality requirements, food additives, contaminants, hygiene, packaging, labelling, methods of sampling and test for Fresh cassava leaves of Manihot esculenta Crantz, for preparation before human consumption
- Reference of the measure
- US EAS 780:2012
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- Fresh cassava leaves of Manihot esculenta Crantz
Countries/Regions affected by the measure.
-
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 25 October 2013
- Publication where the measure is specified
- Uganda Gazette Vol. CV No. 54 of 25th October 2013
- Regulation where the measure is specified
- US 952:2013, Amaranth grain — Specification
- Country/Region applying the measure
- Uganda
- Description of the measure
- This Uganda Standard specifies quality requirements, contaminants, hygiene, packaging, labelling, methods of sampling and test for whole grains obtained from Amaranthus caudutus, A. hypochondaricus and A. cruentus intended for human consumption.
- Reference of the measure
- US 952:2013
- Measure also domestic
- Yes
Products affected by the measure.
- Description
- whole grains obtained from Amaranthus caudutus, A. hypochondaricus and A. cruentus
Countries/Regions affected by the measure.
-
Applied by South Africa on the entire world for 6506.10.90: -- Other
The measure came into effect on 08 September 1984
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 08 September 1984
- Publication where the measure is specified
- Government Notice 1135 (Government Gazette No. 9247) Of 8 June 1984
- Regulation where the measure is specified
- Compulsory Specification For Safety Helmets For Motor Cyclists
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This specification covers requirements for the general design, construction, performance, marking, and labelling,
and testing of safety helmets for use by motor cyclists on the roads - Coded list of objectives
- X: For purposes n.e.s.
- Description of the measure
- 7.2 Conditioning
7.2.1 Pre-conditioning: Expose the helmet to a temperature of 24 +/- 2 °C and a relative humidity of 45+/-5 % for at least six hours.
7.2.2 High temperature
(a) Apparatus: An oven having fan- operated air circulation, an internal volume of at least 0,15m3 and not less than 0,06 m3 per helmet, and thermostatically controlled to maintain a temperature of 50+/- 2°
C .
(b) Procedure: Place the helmet in the oven ensuring hat it does not touch the side , top, or bottom of the oven. Expose the helmet to the temperature of 50+/- 2°C for not less than four hours and not more than five hours.
7.2.3 Low temperature:
(a) Apparatus: A reasonable airtight refrigerator having an internal volume of at least 0,15m3 and not less than 0,06m3 per helmet , and that is thermostatically controlled to maintain a temperature of 0+/- 3°C.
(b) Procedure : Place the helmet in the refrigerator , and expose the helmet to the temperature of 0+/- 3°C for not less than four hours and not more than five hours.
7.2.4 Ultraviolet radiation and water immersion:
(a) Apparatus:
(1) A U.V. quarts lamp of power 125W.
(2) A vessel of adequate size containing water at a temperature of 24 +/- 2°C.
(b) Procedure:
(1) Expose for 48 hours the outer surface of the helmet to ultraviolet radiation from the lamp placed at a distance of 250mm from the helmet.
(2) Invert the helmet and immerse it totally in the water in the vessel for not less than four hours and not more than five hours.
7.3 Verification of Extent of protection and peripheral vision
7.3.1 Apparatus: A headform of appropriate size (see 8.4)
7.3.2 Procedure:
(a) Place the helmet on the headform and secure it firmly using the alignment of the edge above the face in relation to the basic plan at the front and sides of the headform to level the helmet.
(b) Check the extent of the protection of the helmet visually against the circumference AA1 and the lines of CDEF marked on the headform (see Fig 3).
c) Check the extent of peripheral vision (see 4.6) .
(d) Verify compliance with the requirements of 4.6, 5.1(a) and 5.1 (b) .
e) Note the vertical distance between the edge of the helmet above the face and he basic plane , measured at the froth midpoint of the helmet (see 7.4.3 (a) )
7.4 Shock Absorption test
7.4.1 (a) Headform: A headform of appropriate size (see 8.4)
(b) Mounting: The apparatus is mounted on a rigid monolithic base having a mass of at least 500kg. The headform is secured to a spigot coincident with its central vertical axis and the spigot is mounted on an arc that pivots about a horizontal axis (see Fig 4) . The position of the headform along the arc is adjusted to the angle required for the inclined axis of the headform. Rotation of the headform about the spigot presents any selected point on the circumference of the headform to the striker. A similar spigot secured directly to the rigid base enables the headform to be mounted vertically for the delivery of blows to the top of the helmet.
c) Striker: The striker has a mass of 5 kg and a flat circular striking face of a diameter 125 +/- 1,0 mm or a hemispherical striking face of radius 50+/- 0.5 mm, and is dropped in guided fall with minimal retardation from the guides , which are vertical to within 1 in 400.
(8) Acceleration transducer: An acceleration transducer capable of withstanding a 20 000 m/s2 shock without damage is firmly attached to the striker with its vertical axis coincident, to within +/- 2°C, with that of the striker.
(e) Measuring system: The measuring system including the striker has aflat response within +/- 1dB from 5 Hz to 3 kHz.
7.4.2 Instrumentation check: Before commencing helmet tests, check the measuring system by impacting a suitable test piece with the hemispherical striker, dropping the striker from an established height to produce an acceleration of 4 000 m/s2. Record at least three such impacts on each occasion of checking and check that the results lie within a range of 400 m/s2.
7.4.3 Procedure: Use any one of the test schedules given in Table 2 and after subjecting the helmet under test to the appropriate conditioning, proceed as follows:
(a) Securely fasten the helmet to the headform with the required impact site of the helmet presented to the striker, and levelled with the front edge of the helmet set at the same distance from the basic plane at the front midpoint of the headform as previously measured in 7.3.2 (e).
(b) determine ted the mass load of the helmeted headform abut the pivot shaft and in the vertical axis of the striker.
(c) Calculate the height through which the striker has to be dropped to attain an impact energy of 122J in the case of the flat striker and 88 J in the case of the hemispherical striker. These impact energies are attained by the striker falling through a height of (k+1h)/k metres ,
Where k = mass of helmeted headform, kg/ mass of striker, kg
h =2,5 m for flat striker and 1,8 m for hemispherical striker
(d) Raise the striker to the required height, measured from the underside of the striker to the helmet shell, and ensure that it is accurate to within 5 mm.
(e) Drop the striker onto the helmet and record the acceleration.
(f) After any necessary repositioning of the helmet on the hedform, and ensuing that the striker drops on the same impact site on the helmet, repeat the procedure given in (a) – (e) above.
(g) Test the helmet on three sites separated by distances of not less than one-fifth of the maximum circumference of the helmet.
(h) Ensure that side impacts are between 0 mm and 25 mm rearwards of the transverse plane through the central vertical axis of the headform, and on line BB1, i.e. the line generated when the angle between the inclined and horizontal plane of the headform is 20° (see Fig. 1 ).
(i) Ensure that front and rear impacts are within 25 mm of the central longitudinal axis of the headform, and on line BB1.
(j) Verity compliance with the requirements of 5.2 (a).
7.5 Penetration test
7.5.1 Apparatus: The apparatus (see Fig. 5) consists essentially
of the following:
(a) A hemispherical test block of hardwood and having a soft metal insert at the top of its central axis and mounted on arigid base.
(b) A striker with the following characteristics :
Mass .................................. 3,0kg (+45/-0g)
Angle of cone .......................... 60°+/- 0,5°
Height of cone.. ....................... 40 mm, mh.
Radius of point ........................ 0,5 +/- 0,lmm
Hardness of tip ........................ 45-50 HR
(c) A method of electrically indicating when the striker touches the soft metal insert.
7.5.2 Procedure:
(a) Fasten the helmet securely to the test block.
(b) Allow the striker to fall freely from a height 2,5 mm +/- 5mm (measured from the point of the striker to the anticipated point of impact on the helmet) onto the helmet at
two sites on or above the circumference BB1 (see 7.4.3 (h)) and separated by at least 45mm from each other and from the centres of the impact sites of the shock absorption test.
c) Note whether contact is made between the striker and the soft metal insert and, if necessary, restore the surface of the latter, before the next test.
d) Verify compliance with the requirements of 5.2 (b)
7.6 Test for strength of retention system
7.6.1 Apparatus : The apparatus (see Fig 6) consists essentially of the following:
(a) a headform of appropriate size
(b) A mount for the headform that incorporates a load bearing support for the brim of the helmet.
c) A stirrup that has two metal rollers, each 12,5 +/- 0,5 mm in diameter , spaced at 75,0 +/- 0,5 mm centres.
(d) A guide bar and anvil integral with and extending vertically downwards from the stirrup and on which is mounted a gauge for measuring both the maximum dynamic and residual extension of the chin strap. The system has a total mass of 7,0 kg (+0.25/-0) hat is carried on the guide bar and that can be dropped in free fall onto the anvil, which carries a 10mm thick pad of polyethylene foam.
7.6.2 Procedure:
(a) So secure the helmet over the headform and onto the load-bearing support that any load placed on the fastened chin strap is borne by the support and not by the headform, which acts only as a positioning guide for the helmet.
(b) so fasten the chin strap under the stirrup that the chin strap support the stirrup and the guide bar and the anvil system, and that the stirrup is 120 – 140 mm below the headform.
(c) Adjust the extension measuring gauge to zero.
(d) Drop the 10 kg mass piece from a height of 750 +/- 5mm onto the anvil.
e) Record the dynamic extension and with the 10 kg mass still on the anvil, measure the residual extension
(f) Reset the extension –measuring gauge to zero
(g) Without touching the chin, strap repeat (d) and e)
(h) verify compliance with the requirements of 5.3
7.7 Test for rigidity
7.8 Corrosion resistance test
7.9 Test for flexibility of peak
7.10 Test for strength on visor - Reference of the measure
- Regulations 7.2 to 7.10
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 6506.10.90 -- Other Yes Helmets for motor cyclists - Description
- Helmets for motor cyclists
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 3808.94: -- Disinfectants
The measure came into effect on 14 July 1999
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 14 July 1999
- Publication where the measure is specified
- Government Notice R529 (Government Gazette 19999) Of 14 May 1999
- Regulation where the measure is specified
- Compulsory Specification For Disinfectants And Detergent-Disinfectants
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This specification covers requirements for disinfectants and detergent-disinfectants intended for use on inanimate surfaces.
- Coded list of objectives
- X: For purposes n.e.s.
- Description of the measure
- 5 Methods of microbiological testing
5.1 General
The tests shall be undertaken by persons experienced in microbiological techniques, using aseptic techniques.
NOTES
1 In order to ensure accuracy of these tests, it is recommended that each test be repeated.
2 Before these tests are carried out, the efficacy of the Inactivating system should be checked to ensure that it adequately inactivates the disinfectants or detergent-disinfectants to be tested.
3 For the purpose of checking the resistance of the test organisms and the other test conditions, it is advisable to include a reference standard. It is essential that it be a disinfectant or detergent-disinfectant based on the relevant active ingredient,
but, because a universal standard is difficult to select, each laboratory should make its own choice of material.
5.2 Laboratory ware
Ensure that all glassware is resistant to repeated heat sterilization and that the glass is free from inhibitory substances such as heavy metals and free alkalis. Borosilicate glass with an expansion coefficient of less than 6 x 10^(-6%)k^(-1) is recommended.
5.2.1 Universal container culture bottles
Bottles made of glass, fitted with standard screwed metal caps with rubber liners and of nominal capacity:
a) 30ml, and
b) 110 ml.
Do not use plastics containers or glass containers fitted with plastics tops.
5.2.2 Culture tubes
Rimless cylindrical tubes that have hemispherical ends and a nominal wall thickness of 1,5 mm and that are of the following sizes:
a) diameter and length 16 mm x 160 mm; and
b) diameter and length 20 mm x 200 mm.
Plug these tubes with cotton wool plugs or with plugs of a foam rubber suitable for autoclaving or use screw-capped tubes of similar dimensions.
5.2.3 Graduated pipettes
Total delivery pipettes for bacteriological purposes only, that have an outflow opening of diameter 2 mm to 3 mm, are graduated in units of 0,1 ml and are of sizes to deliver 1 ,0 ml 5,0 ml and 10,0 ml.
5.2.4 Volumetric cylinders
Graduated measuring cylinders with or without stoppers and of capacities 5 ml. 1 ml, 1 00 md, 500 ml and 1 000 ml.
5.2.5 Culture flasks
Culture flasks of capacities 250 m«, 500 ml and 1l
5.2.6 Erienmeyer flasks
Erienmeyer flasks of capacities 250 ml, 2 l and 3 l.
5.2.7 Petri dishes
Petri dishes of diameter and height 100 mm x 20 mm and made of glass or of wettable polystyrene.
5.2.8 Reagent bottles
Bottles of capacities 50 ml and 1 00 m« and that have polypropylene or other plastics stoppers of such design that they can be used to deliver drops of the reagent.
5.2.9 Inoculating loop
A length of platinum or platinum-iridium alloy wire of diameter 0.376 mm, mounted in a holder that consists of a thin metal rod or tube. The end of the wire is formed into a loop of diameter 4 mm at a distance of 38 mm from the holder. The loop is at such an angle to the axis of the wire that it can be kept in the horizontal plane while being lifted vertically off the surface of the liquid.
5.2.10 Droplet pipette
A droplet pipette that delivers 0,2 mi in about five droplets.
5.2.11 Micropipetter
A micropipetter, high precision, adjusted to dispense 20 \ii accurately, and sterile tips suitable for use with this micropipetter.
5.3 Equipment
5.3.1 Autoclave
A pressure vessel capable of producing steam or connected to a central steam source and capable of withstanding a pressure of 300 kPa. The autoclave is capable of attaining a temperature of 121 °C within 10 min of the beginning of the sterilization cycle.
5.3.2 Incubators and water-baths
Incubators and water-baths that have thermostatically controlled heating and cooling devices and that are so fitted with means of circulation that the temperature of the total enclosed space is maintained to within 2 °C of the thermostat setting.
5.3.3 Hot air oven (for sterilization by means of dry heat)
A thermostatically controlled oven heated by electricity or gas and so fitted with means of circulation that the temperature of the total enclosed space is maintained at 1 70 °C ± 5 °C, the heat supply being such that the working temperature is regained within 10 min of the momentary opening and closing of the oven door.
5.3.4 Stop-watch
A stop-watch accurate to within 1 s per hour.
5.4 Media and reagents
5.4.1 General
5.4.1.1 Water
Use only glass-distilled water or demineralized water of equivalent purity, that is clear, colourless and free from visible suspended matter and of which the pH value, measured at 25 °C, is in the range 5,0 to 7,5.
5.4.1.2 Quality of ingredients
In the preparation of the media and reagents, use only ingredients of quality acceptable for microbio- logical purposes. Use anhydrous salts unless otherwise specified.
5.4.1.3 Accuracy
Except where otherwise specified, allow the following tolerances:
a) on temperatures ………..± 2 °C
b) on masses ………………± 1 ,0 %
c) on volumes………………….. ± 1 ,0 %
d) on pH value………………………………….. ±0,1
5.4.1.4 Dehydrated media
Many of the media required are obtainable in dehydrated form and, for uniformity of results, the use of such media is recommended. If these are used, follow the manufacturer's instructions strictly with regard to reconstitution and sterilization.
5.4.1.5 Filtration Of media
Whenever it is necessary to filter a medium in the course of its preparation, proceed as follows:
a) filter a medium that does not contain a solidifying agent, i.e. a liquid medium or broth, through a medium-speed filter paper; or
b) if the medium contains a solidifying agent (for example, agar) filter it through al0mm to15mm thick layer of pre-wetted absorbent cotton wool. To prevent solidification of the medium during filtration, use a steam-jacketed funnel. Alternatively, carry out the filtration in a steam chamber.
5.4.1 .6 Adjustment of the pH value of media
Where the final pH value of a medium or reagent is specified, so adjust the pH value, if necessary, during preparation and, in the case of media, before sterilization, that, after preparation, the required pH value measured at 25 °C is obtained. Unless otherwise specified, use a solution of hydrochloric acid (c(HCI)
= 1 mol/l) or sodium hydroxide (cNaOH) = 1 mol/l), as appropriate, to adjust the pH values.
5.4.1.7 Dispensing
Where specified quantities of media are to be dispensed into bottles, use 30 mi universal bottles (see 5 2.1(a)). Where bulk sterilizing is required, use any suitable glass container of the required quality or suitably stoppered culture tubes (see 5.2.2(a)). Dispense reagents into reagent bottles (see 5.2.8). Stir media constantly while dispensing. Whenever the preparation of slopes for surface cultivation is required,
dispense the medium in 10 m« volumes and sterilize as specified. Immediately after sterilization, place the bottles or, when relevant, the culture tubes, on a 1 -in-4 sloped surface and allow the agar to solidify.
5.4.1.8 Sterilization
When sterilization by autoclaving is specified and unless otherwise directed, autoclave the medium at 121 °C for 15min.
5.4.1.9 Control of prepared media
Ensure, by suitable incubation tests, that prepared media are sterile and are capable of supporting the growth of the relevant organisms under the stated conditions of incubation.
5.4.1.10 Storage Of media
Ensure that prepared media are carefully protected from exposure to heat and sunlight and have not evaporated or changed in concentration or in pH value, and that, unless otherwise specified, they are used within three months of preparation.
5.4.2 Bottom layer agar
5.4.2.1 Ingredients
Tryptone ………………13.0g
Agar………………… 11,0g
Sodium chloride…………….. 8,0g
Glucose…………… 1,0g
Water………………………. 1 000 ml
5.4.2.2 Preparation
Dissolve the ingredients in the water by warming. Cool to 45 °C to 50 °C and dispense into Petri dishes of nominal diameter 90 mm, ensuring that the depth of the agar in each plate is at least 3 mm.
5.4.3 Bovine albumin solution
5.4.3.1 Ingredients
Albumin (bovine) ………………………….1 5,0 g
Water……………………………. 1 000 ml
5.4.3.2 Preparation
Dissolve the albumin in the water. Sterilize by passing through a filter with maximum effective pore size of 0,45 µm. Dispense 10 ml volumes into bottles (see 5.2.1(a)).
5.4.4 Cetrimide inactivator
5.4.4.1 Ingredients
Polyoxyethylene sorbitan mono-oleate…………….. 8,0 g
Sodium taurocholate…………… 8,0 g
Sodium thiosulfate…………..1.5 g
Potassium phosphate, monobasic………………. 0,5 g
Sodium citrate…………… 0,5 g
Water…………………. 1 000 ml
5.4.4.2 Preparation
Dissolve the ingredients in the water by heating. Dispense 20 ml volumes into bottles (see 5.2.1 (a)) and sterilize by autoclaving.
5.4.5 Diluent used in the "5,5,5" test
5.4.5.1 Ingredients
Albumin (bovine)………………………. 0,3 g
Sodium chloride………………………………………. 9,0 g
Water ……………………………..1 000 ml
5.4.5.2 Preparation
Dissolve the ingredients in the water. Sterilize by passing through a filter with maximum effective pore size of 0,45 µm. Dispense 10 ml volumes into bottles (see 5.2.1(a)).
5.4.6 Hard water used in preparing the yeast suspension
5.4.6.1 Ingredients
Calcium chloride (CaCl2) ……………..1 ,0 g
Magnesium chloride (MgCl2-6H2O)………………………… 8,5 g
Water …………………………………..1 000 ml
5.4.6.2 Preparation
Dissolve the ingredients in the water. Dispense 1 00 ml volumes into bottles {see 5.2.1 (b)) and sterilize by autoclaving.
5.4.7 Inactivator media
5.4.7.1 Inactivator medium No. 1
5.4.7.1.1 Ingredients
Monopotassium phosphate (KH2PO4) …………………0,5 g
Sodium citrate (Na3C6H5O7.3H2O)……………………………. 0,5 g
Sodium taurocholate……………………………….. 8,0 g
Sodium thiosulfate (Na2S2O3.5H2O)……………………… 1 .5 g
Polyoxyethylene sorbitan mono-oleate ………………………….8,0 g
Water……………. 1 000 ml
5.4.7.1.2 Preparation
Dissolve the ingredients in the water by heating. Dispense 20 mi volumes into bottles (see 5.2.1 (a)).
Sterilize by autoclaving.
NOTE - This inactivator has been found suitable for disinfectants and detergent-disinfectants based on iodopliors, organic halogen compounds {other than iodine compounds) and most quaternary ammonium compounds.
5.4.7.2 Inactivator medium No. 2
5.4.7.2.1 Ingredients
Polyoxyethylene sorbitan mono-oleate ……………30,0 g
Beef extract…………………. 20,0 g
Peptone………………………. 20,0 g
Sodium chloride……………………………..1 0,0 g
Water……………………….. 1 000 ml
5.4.7.2.2 Preparation
Dissolve the ingredients in the water and adjust the pH value to 7.1 . Dispense 9 md and 10 md volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving.
NOTE -This inactivator has been found suitable for disinfectants and detergent-disinfectants based on glutaraldehyde.
5.4.7.3 Inactivator medium for the "5,5,5" test
5.4.7.3.1 Ingredients
Lecithin (made from soya, purified)………………………… 3,0 g
l-histidine………………………………1,0g
Phosphate buffer 0,25N (see 5.4.12)……………………… 10,0 ml
Sodium thiosulfate (Na2S2O3.5H2O)………………… 5.0 g
Polyoxyethylene sorbitan mono-oleate…………………………….. 30.0 ml
Water………………………………………….. 1 000 ml
5.4.7.3.2 Preparation
Dissolve the ingredients in the water by heating. Dispense 20 me volumes into bottles (see 5.2.1 (a)).
Sterilize by autoclaving.
5.4.7.4 Neutralizer broth medium
Use TSB (see 5.4.22) but to each 100 ml of broth, add 3 g of soy lecithin (azolectin) and 20 g of polyoxyethylene sorbitan mono-oleate. When testing an antiseptic that contains cetrimide, also add 20 ml of cetrimide inactivator (see 5.4.4). Adjust the pH value to between 7,0 and 7,4. Dispense 9 ml volumes into bottles (see 5.2.1 (a)) and sterilize by autoclaving for 20 min.
NOTES
1 In order to dissolve the azoiectin and polyoxyethylene sorbitan mono-oleate in the TSB, use half the volume of TSB in a large container and add the azoiectin and polyoxyethylene sorbitan mono-oleate while stin-ing. Boll for 30 min to 60 min with constant stirring until all the azoiectin granules are dissolved. Allow to cool before diluting to the final volume with TSB.
Adjust the pH value and dispense as required. Do not fill the bottles to more than half their volume before sterilizing, to prevent boiling over of the contents, which will alter the azolectin/polyoxyethylene sorbitan mono-oleate ratio.
2 Sometimes the azolectin/polyoxyethylene sorbitan mono-oleate emulsion settles to the bottom of the container. If this
happens, storage of approximately one week at room temperature usually allows the solids to redissolve. If the neutralizer is required sooner, healing in a water-bath at 100 °C followed by occasional swirling during cooling will redissolve the solids.
3 This inactivator has been found suitable for antiseptics based on chlorhexidine gluconate and for some disinfectants and detergent-disinfectants based on quaternary ammonium compounds and glutaraldehyde.
5.4.8 Mait extract agar
5.4.8.1 Ingredients
Malt extract………………………….. 30,0 g
Agar……………………………………. 15,0 g
Soya peptone……………………………….. 5,0 g
Water …………………………….1 000 m
5.4.8.2 Preparation
Dissolve the ingredients in the water. Dispense 10ml and 1 5 ml volumes into bottles (see 5.2.1 (a)) and sterilize by autoclaving. Allow only the 10 ml volumes to solidify in a sloped position.
5.4.9 Nutrient agar
5.4.9.1 Ingredients
Agar…………………………….. 15,0 g
Peptone……………………………….. 5,0 g
Sodium chloride …………………………………5,0 g
Yeast extract …………………………….2,0 g
Beef extract……………….. 1,0 g
Water …………………………1 000 ml
5.4.9.2 Preparation
Dissolve the ingredients in the water and adjust the pH value to 7,1 . Dispense 1 m{ and 15 ml volumes into bottles (see 5.2.1 (a)) and sterilize by autoclaving. Allow only the 1 ml volumes to solidify in a sloped position.
5.4.10 Nutrient broth No. 2 (double strength)
5.4.10.1 Ingredients
Peptone………….. 10,0 g
Sodium chloride …………………………5,0 g
Refined meat extract ……………….10,0 g^1
Water ………………………………..1 000 ml
5.4.10.2 Preparation
Dissolve the ingredients in water, adjust the pH value to 7,1 and dilute the solution to 1 t Dispense 5 ml volumes into culture tubes (see 5.2.2(a)) and sterilize by autoclaving.
5.4.11 Nutrient medium
5.4.11.1 Ingredients
Peptone…………………….. 5,0 g
Sodium chloride…………………………….. 5,0 g
Yeast extract ………………………………2,0 g
Beef extract ………………………………1 ,0 g
Water…………………………. 1 000 ml
5.4.11.2 Preparation
Dissolve the ingredients in the water and adjust the pH value to 7,1 , Dispense 10 mi and 50 mi volumes into bottles (see 5.2.1(a) and 5.2.1(b), respectively) and sterilize by autoclaving.
5.4.12 Phosphate buffer 0,25N
5.4.12.1 Ingredients
Monopotassium phosphate (KH2PO4)………………………. 34,0 g
Water ……………………………………….500 ml
5.4.12.2 Preparation
Dissolve the monopotassium phosphate in the water. Adjust the pH value to 7,2 with 1 N NaOH. Dispense into 30 m{ bottles (see 5.2.1(a)) and sterilize by autoclaving.
5.4.13 Physiological saline solution
5.4.13.1 Ingredients
Sodium chloride……………………………… 9.0 g
Water…………………. 1 000 ml
5.4.13.2 Preparation
Dissolve the sodium chloride in the water. Dispense into 250 mi culture flasks (see 5.2.5) and sterilize by autoclaving.
1) Meat extract made from specially selected raw materials of a light colour adjusted to neutrality and dried to a fine powder.
5.4.14 Recuiture medium
5.4.14.1 Ingredients
Beef extract………………………. 10 g
Peptone………………………………… 1 g
Sodium……………………… chloride 5 g
Polyoxyethylene sorbitan mono-oleate………………………….. 30 g
Water…………………………… 1 000 ml
5.4.14.2 Preparation
Dissolve the ingredients in the water and adjust the pH value to 7,5. Dispense 1 mfi volumes into culture tubes (see 5.2.2(a)) and sterilize by autoclaving.
5.4.15.1 Sodium thiosulfate (20 g/H)
5.4.15.1 ingredients
Sodium thiosulfate…………………………… 20 g
Water……………………………… 1 000 ml
5.4.15.2 Preparation
Dissolve the sodium thiosulfate in the water, dispense 20 m? volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving.
NOTE - This Inactivator has been found suitable for disinfectants and detergent-disinfectants based on stabilized inorganic chlorine compounds arid stabilized chlorine compounds.
5.4.16 Sodium thiosuifate (10 g/e)
5.4.16.1 Ingredients
Sodium thiosulfate………………….. 1 g
Water ………………………………..1 000 ml
5.4.16.2 Preparation
Dissolve the sodium thiosulfate in the water, dispense 20 ml volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving.
5.4.17 Sporulation medium SM 1
5.4.17.1 Ingredients
Agar…………………… 12,0 g
Manganese sulfate (MnS04.4H20)…………………………… 0,03 g
Dipotassium phosphate…………………………. 4,0 g
Nutrient broth …………………………………3,1 25 g
Water…………………………… 1 000 ml
5.4.17.2 Preparation
Dissolve the ingredients in the water and adjust the pH value to 6,6. Dispense 20 mS. volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving. Allow the medium to solidify in a sloped position.
5.4.18 Standard hard water
54.18.1 Standard hard water No. 1
5.4.18.1.1 Ingredients
Calcium chloride……………………….. 2,1 g
Water………………………………. 7 500 ml
5.4.18.1.2 Preparation
Dissolve the calcium chloride in the water. Dispense 97 ml volumes into 1 10 m« bottles (see 5.2.1(b)) and sterilize by autoclaving.
NOTE -This hard water has been found suitable tor the microbiological testing of disinfectants and detergent-disinfectants based on glutaraldehyde, organic halogen compounds (other than Iodine compounds), quaternary ammonium compounds, stabilized inorganic chlorine compounds and stabilized chlorine compounds.
54.18.2 Standard hard water used for the microbiological testing of disinfectants and detergent- disinfectants based on iodophors
5.4.18.2.1 Ingredients
Magnesium chloride………………………… 1 8,5 g
Calcium chloride……………………………. 7,9 g
Sodium bicarbonate…………………………… 22,4 g
Water……………………………. 2 500 ml
5.4.18.2.2 Preparation
Dissolve the magnesium chloride and calcium chloride in water, adjust the pH value to between 7,6 and 8,0 and dilute to 1 « with water. Sterilize by autoclaving (solution A). Dissolve the sodium bicarbonate in water, adjust the pH value to between 7,6 and 8,0 and dilute to 1 8 with water. Sterilize by filtration (solution B). Add 1 me of each of solutions A and B to 95 ml. of sterile water in an Erlenmeyer flask (see 5.2.6).
5.4.18.3 Standard hard water for use in the Kelsey-Sykes test
5.4.18.3.1 Ingredients
Calcium chloride (CaCl2)………………………… 0.304 g
Magnesium chloride (MgCl2.6H2O) ……………………..0,139 g
Water …………………………..1 000 ml
5.4.18.3.2 Preparation
Dissolve the ingredients in the water. Dispense 100 md volumes into 1 10 mi bottles (see 5.2.1 (b)) and sterilize by autoclaving.
5.4.18.4 Standard hard water used in the Rideal- Walker test
5.4.18.4.1 Ingredients
Calcium chloride ……………………..0,15 g
Magnesium sulfate…………………….. 0,15 g
Water……………. 1 000 ml
5.4.18.4.2 Preparation
Dissolve the ingredients in the water. Dispense 100 ml volumes into 110 ml bottles (see 5.2.1(b)) and sterilize by autoclaving.
5.4.19 Standard phenol solution (50 g/l)
Using pure phenol that has a crystallizing point not lower than 40,5 °C, prepare a 50 g/l stock solution in water and use this for making the control dilutions (see 5.10.2).
NOTE - It is important that pure phenol be used, because cresol has approximately three times the bactericidal efficacy of phenol, and error resulting from its presence (as indicated by reduction of the crystallizing point to below 40,5 °C) might be considerable.
5.4.20 Sterile skimmed milk
5.420.1 Blend 50,0 g of skimmed milk powder with 400 ml of water in a high-speed blender for 5 min to 7 min. Dilute to 500 ml with water.
5ASi02 Filter through a coarse filter paper that has been previously wetted, or centrifuge for 10 min at a resultant centrifugal force of 6 kN/kg. If there is a visible residue on the filter paper or in the centrifuge tube, repeat the procedure with a different batch of skimmed milk.
5.4.20.3 Dispense 15 ml to 25 ml volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving for 10 min. Store in a refrigerator maintained at 4 °C.
5.4.20.4 Test for freedom from growth-Inhibiting factors
5.4.20.4.1 Using sterile water, prepare a suspension of Staphylococcus aureus (see 5.5.2.1). So standardize the suspension, by using a spectrophotometer in conjunction with a standard curve, a haemocytometer, Petroff-Hausser countina chamber or any other suitable means, that it contains approximately 100 million organisms (10*) per milliiitre. Use the suspension within three hours of preparation.
5.4.20.4.2 Mix 0,12 ml of this suspension with 15 ml of melted nutrient agar (see 5.4.9) that has been cooled to 45 °C and pour the mixture into a flat-bottomed Petri dish of diameter 100 mm (see 5.2.7), placed on a level surface. After the agar has set, place the Petri dish for at least 15 min in a refrigerator maintained at 2 °C to 5 °C. Put three sterile, polished, stainless steel cylinders (penicillin cups) of outer
diameter 8 mm ± 0,5 mm, inner diameter 6 mm ± 0,5 mm and length 10 mm ± 1 mm on the agar and fill each with the milk to be tested.
5.4.20.4.3 Leave the Petri dish in the refrigerator for 2 h and then incubate the Petri dish at 37 °C for 1 6 h to 20 h. Remove the cylinders from the Petri dish and examine the set agar plate for zones of growth inhibition.
5.4J20.4.4 Repeat the procedure given in 5.4.20.4.1 to 5.4.20.4.3 (inclusive), using:
a) Escherichia coli as the test organism , with:
1) a suspension containing 650 million (6,5 x 10®) organisms per milliiitre; and
2) an inoculum of 0,15 ml in 15 ml of melted nutrient agar; and
b) Pseudomonas aeruginosa as the test organism, with:
1) a suspension containing 500 million (5,0 x 10®) organisms per milliiitre; and
2) an inoculum of 0,15 ml in 15 ml of melted nutrient agar.
5.4.20.5 If no zones of growth inhibition are visible on any of the three sets of plates, the milk may be used for the test.
5.4.21 Top layer agar
5.4.21.1 Ingredients
Tryptone……………………………..1 0,0 g
Sodium chloride………………………… 8,0 g
Agar ………………………………………….6,0 g
Glucose………………………….. 3,0 g
Water ………………………..1 000 ml
5.4.21.2 Preparation
Dissolve the ingredients in the water by warming. Dispense 90 ml volumes into 110 ml
bottles (see 5.2.1 (b)) and sterilize by autoclaving.
5.4.22 Tryptone soy broth (TSB)
5.4.22.1 Ingredients
Tryptone …………………………….17,0 g
Sodium chloride………………………………….. 5,0 g
Soy peptone……………………………………………… 3,0 g
Potassium hydrogen dibasic-phosphate……………………………………… 2,5 g
Dextrose…………………………………. 2,5 g
Water ……………………………1 000 ml
5.4.22.2 Preparation
Dissolve the ingredients in the water, heating if necessary. Adjust the pH value to 7,3, dispense 10 ml volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving.
5.4.23 Wright and Mundy medium (synthetic broth AOAC)
5.4.23.1 Part A
5.4.23.1.1 Ingredients
l-cystine………………… 0-05 g
dl-methionine……………………………….. 0,37 g
l-arginine hydrochloride………………………… 0,4 g
dl-histidine hydrochloride………………………. 0,3 g
l-lysine hydrochloride…………………… 0,85 g
l-tyrosine………………………………… 0,21 g
dl-threonine………………………………… 0,5 g
dl-valine…………….. 1,0 g
l-leucine ……………………………0,8 g
dl-isoleucine ……………………………….0.44 g
Glycine…………………………. 0,06 g
dl-serine…………………………………… 0,61 g
dl-alanine………………………. 0,43 g
l-glutamic acid hydrochloride ……………………………..1 .3 g
l-aspartic acid …………………………………………..0,45 g
d l-phenylalanine………………………………. 0,26 g
dl-tryptophan……………………………… 0,05 g
l-proline …………………………0,05 g
Water ……………………………….500 ml
5.4.23.1.2 Preparation
Dissolve the ingredients in the water and add 1 8 m{ of a sodium hydroxide (c(NaOH) = 1 mol/l) solution.
5.4.23.2 Parts
5.4.23.2.1 Ingredients
Sodium chloride ………………………………..3,0 g
Potassium chloride……………………………………… 0,2 g
Magnesium sulfate………………………………………….. 0,05 g
Potassium phosphate……………………………………… l,5 g
Disodium phosphate …………………………………4,0 g
Thiamine hydrochloride……………………… 0,01 g
Nicotinamide ………………………0,01 g
Water ……………………………..500 ml
5.4.23.2.2 Preparation
Dissolve the ingredients in the water. Mix Parts A and B. If necessary, adjust the pH value to 7,1 . Dispense half of the medium in 10 m{ ± 0,2 mi volumes and the other half in 6 m{ ± 0,2 mi volumes into bottles (see 5.2.1(a)) and sterilize by autoclaving.
Before use, add to each tube in the two sets of bottles 0,1 me and 0,06 me, respectively, of a sterile 100 g/l solution of glucose.
5.4.24 Yeast suspension
5.4.24.1 Preparation of 20 % (by mass) of moist yeast suspension
NOTES
1 Whenever possible, the 20 % (by mass) of moist yeast suspension should be sterilized on the day the yeast is received. If this is not feasible, the unopened yeast package should be stored at a temperature not higher than 5 °C for not longer
than 48 h before use.
Crumble approximately 500 g of baker's yeast by hand into a previously tared 1 l beaker and determine the mass of the moist yeast. Cream the yeast by adding a small volume of hard water (see 5.4.6) while stirring the mixture. Carefully transfer the creamed portion to an Erienmeyer flask of capacity 2 i (see 5.2.6) and add a further small volume of hard water to any lumpy residue remaining in the beaker.
Continue this process until all the yeast has been transferred from the beaker to the flask and the concentration of the yeast suspension in the flask has been reduced to approximately 40 % (by mass) of moist yeast. Shake the contents of the flask vigorously and remove large particles by passing the suspension through a sieve of aperture size 140 pm, supported in a funnel in an Erienmeyer flask of capacity 3 l (see 5.2.6). Add enough hard water to reduce the concentration of the yeast to approximately 20 % (by mass) of moist yeast. Shake thoroughly and, while agitating, dispense 100 ml
volumes into bottles (see 5.2.1 (b)). Sterilize by autoclaving and store at 4 °C until required for use.
5.4.24.2 Determination of moisture content
Pipette 25 ml of the sterilized yeast suspension (see 5.4.24.1 ) into a dry tared
dish and dry to constant mass in a hot air oven maintained at 100 °C. Use this mass to determine the additional volume of hard water that must be added to each bottle of sterilized yeast suspension to make a suspension that contains exactly 5 % (by mass) of dry yeast.
5.4.24.3 Adjustment of pH value
Using a sodium hydroxide (c(NaOH) = 1 mol/«) solution, adjust the pH value of 1 00 ml of the 20 % (by mass) of moist yeast suspension (see 5.4.24.1) to 7,0, and note the volume of the sodium hydroxide solution required.
5.4.24.4 Preparation of 5 % (by mass) of dry yeast suspension
Immediately before use add to 100 ml of 20 % (by mass) of moist yeast suspension (see 5.4.24.1), the volume of hard water (see 5.4.6) necessary to mal<e a suspension that contains exactly 5 % (by mass) of dry yeast and enough sodium hydroxide (c(NaOH) = 1 mol/fi) solution (see 5.4.24.3) to adjust the pH value to 7,0. Store the yeast suspension at 4 "C for not longer than 7 d before use.
5.5 Test organisms
Use the following test organisms:
a) Staphylococcus aureus SABS TCC Sta 53 and SABS TCC Sta 59;
b) Escherichia coli SABS TCC Esc 25;
c) Escherichia coli K-1 2 Hf r, NCTC1 2486 SABS TCC Esc 36;
d) Escherichia coli 36 with bacteriophage MS2 SABS TCC Phg-C1 in cases where virucidal
e) Escherichia coli ATCC 1 3706 SABS TCC Esc 37;
f) Escherichia coli 37 with bacteriophage <t)X1 74 SABS TCC Phg-C2 in cases where virucidal
g) Pseudomonas aeruginosa SABS TCC Pse 2 and SABS TCC Pse 16;
h). Salmonella typhi SABS TCC Sal 10 (NCTC 786. Lister strain)
i) Aspergillus r)iger SABS TCC 355 in cases where fungicidal
j) Bacillus subtilis var. globigii SABS TCC Bac 35 in cases where sporicidal
Organisms that have survived the action of a disinfectant or detergent-disinfectant shall under no circumstances be used in a test.
Notes
1 Additional organisms may be used if so desired.
2 The extreme importance of using the standard strain is emphasized.
5.5.1 Maintenance of test organisms
5.5.1.1 Escherichia coli. Salmonella typhi. Staphylococcus aureus and Pseudomonas aeruginosa
5.5.1.1.1 From a newly opened freeze-dried culture or recently received agar culture, subculture the test organisms into bottles of 10 ml nutrient medium (see 5.4.11).
(Use tryptone soy broth (see 5.4.22) in the case of disinfectants based on chlorhexidine gluconate.)
5.5.1.1.2 Incubate the bottles at 37 °C for 24 h. Make subcultures from the cultures in the bottles onto slopes of nutrient agar (see 5.4.9). Incubate the slopes at 37 °C for 24 h.
5.5.1.1.3 From each of these slope cultures, prepare four subcultures (stock cultures) of each test organism on 10ml nutrient agar slopes (see 5.4.9). Incubate the stock cultures at 37 °C for 24 h and then store them in a refrigerator maintained at 4 °C.
NOTE -Take not more than six serial subcultures from each stock culture before resorting to a new freeze-dried culture.
5.5.1.2 Aspergillus niger
Inoculate slopes of malt extract agar (see 5.4.8) with Aspergillus niger and incubate at 25 °C for 7 d.
5.5.1 .3 Bacillus subtilis var. globigii
5.5.1.3.1 Inoculate slopes of sporuiation medium Sr\/1 1 (see 5.4.17) with B. subtilis var. globigii and incubate at 30 °C for 7 d and then at ambient temperature until 80 % to 100 % spores are present (i.e. approximately 5 d).
5.5.1 .3.2 Determine the degree of sporuiation microscopically. When 80 % to 1 00 % spores are present, harvest the spores by adding sterile water and gently scraping the agar surface. Alternatively, use a sterile glass rod or sterile beads to rub the spores off.
5.5.1.3.3 Centrifuge the suspension for 20 min to 30 min. Decant the supernatant liquid. Wash the spores 3 times to 4 times with sterile water until the supernatant liquid is completely clear. Re-suspend the spores in sterile water and heat the suspension for 1 min at 80 °C. Cool the suspension rapidly and store at 4 °C for 7 d.
5.5.1.34 Finally, wash the spore suspension once more in sterile distilled water and so standardize the suspension, by using a spectrophotometer in conjunction with a standard curve, a haemocytometer, Petroff-Hausser counting chamber or any other suitable means, that it contains at least 10 million (10 ) spores per millilitre.
5.5.1.3.5 Dispense into sterile bottles (see 5.2.1 (a)) and store the spore suspension at 4 °C until needed.
5.5.2 Preparation of cultures for test suspensions
5.5.2.1 Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa
5.52.1,1 For each of the test organisms, inoculate a nutrient agar slope (see 5.4.9) from a stock culture kept at 4 °C (see 5.5.1 .1 .3) and incubate at 37 °G for 24 h.
5.5.2.1.2 For the test, use a 24 h culture that has been subcultured for two successive days. After six subcultures, restart the process, using a fresh stock culture (see 5.5.1 .1 .3).
NOTE - The physiological condition of the test organisms is important and might influence inter-laboratory and intra- laboratory variations in test results.
5.5.2.1.3 Using 10 ml of sterile water, wash the bacterial growth resulting from 24 h incubation from the slope (see 5.5.2.1.1), scraping the agar surface if necessary. Carefully decant the suspended growth into a sterile Erienmeyer flask (see 5.2.6) and shake vigorously to suspend all growth in the water. So standardize the suspension, by using a spectrophotometer in conjunction with a standard curve, a haemocytometer, Petroff-HSusser counting chamber or any other suitable means, that it contains the
required number of organisms per millilitre as stipulated in the individual microbiological efficacy tests. Use the suspension within three hours of preparation.
5.5.2.2 Salmonella typhi
5.522.1 Transfer a small portion of growth from a stock culture (see 5.5.1 .1 .3) to 5 ml of Nutrient broth No. 2 (double strength) (see 5.4.10) and incubate at 37 °C for 24 h.
5.5.2.2.2 Continue subculturing into fresh tubes of Nutrient broth No. 2 (double strength) at daily intervals, always transferring one standard loopful of the culture. After two weeks of subculturing as described, begin the process again with a fresh stock culture.
SJ522.3 A subculture made on any day between the third day and the fifteenth day (inclusive) may be used for the test, subject to the following provisions:
a) omission of one daily subculturing operation requires no special reorganization of procedure but, if subculturing cannot take place on two successive days, three successive daily subculturing operations shall be carried out after the break before organisms suitable for the test are obtained; and
b) over week-ends, the following procedure may be adopted: on the Friday make a subculture at about 1 0:00 and incubate at 37 °C until 1 6:00. Transfer the subculture to an incubator maintained at 22 °C, and leave it there for the week-end. At 08:00 on the following Monday, transfer it back, for at least 2 h, to the incubator maintained at 37 °C. and then prepare the usual subculture for use on the Tuesday.
NOTE - Discard cultures that show signs of clumping or pellicle formation.
5.5.2.3 Aspergillus niger
Using 10 ml of a sterile 0,5 g/l solution of polyoxyethylene sorbitan mono-oleate, wash the growth resulting from 7 d incubation from the slope (see 5.5.1.2), scraping the agar surface if necessary.
Carefully decant the suspended growth into a sterile bottle (see 5.2.1(a)) and shake vigorously to suspend all growth in the water. So standardize the suspension, by using a haemocytometer, Petroff- Hausser counting chamber or any other suitable means, that it contains the required number of organisms per millilitre as stipulated in the individual microbiological efficacy tests. Use the suspension within three hours of preparation.
5.5.2.4 Bacillus subtilis var. globigii
Prepare the suspension in accordance with 5.5.1 .3.
5.5.2.5 Bacteriophage
NOTE - The preparation refers to bacteriophage MS2 strain (see 5.5(d)). The preparation of bacteriophage (PXI 74 strain is identical, except that a different bacterial culture, i.e. SABS TCC Esc 37, is used.
5.5.2.5.1 Preparation of tiie bacteriopiiage stock
Use either the plate method or the broth method.
5.5.2.5.1.1 Plate method
5.5.2.5.1.1.1 Inoculate a dilution of bacteriophage MS strain that produced a semi-confluent lysis on an agar plate with Escherichia coli SABS TCC Esc 36 into tubes containing top layer agar (see 5.4.21 ) . Pour the suspension onto bottom layer agar (see 5.4.2), and incubate at 37 "C for 18 h.
5.52.5.1.15 Dispense 2,5 ml of nutrient medium (see 5.4.1 1 ) aseptically onto each plate and remove the nutrient medium with top layer agar by scraping it from the bottom layer agar, using a sterile glass rod.
55.2.5.1.1.3 Homogenize the suspension by vigorous shaking and then centrifuge the suspension at 7 000r/min for 10 min.
5.5.2.5.1.1.4 Add 10 % (by volume) of chloroform to the bacteriophages in the supernatant and store this stock solution at 4 °C.
5.5.2.5.1.2 Broth method
5.5.2.5.1.2.1 Add the bacteriophage MS strain to a nutrient medium (see 5.4.11) culture of Escherichia Coll SABS TCC Esc 36 which is near the end of the log phase to ensure that virtually every bacterial cell is infected simultaneously.
5.5.2.5.1.2.2 Centrifuge the suspension at 7 000 r/min for 10 min after lysis has occurred (visible as a marked drop in turbidity).
5.5.2.5.1.2.3 Add 10 % (by volume) of chloroform to the bacteriophages in the supernatant and store this stock solution at 4 °C.
5.5.2.5.2 Titration of the bacteriophage suspension
5.5.2.5.2.1 Inoculate a nutrient agar slope from a stock culture of Escherichia coli SABS TCC Esc 36 (see 5.5.1 .1 .3) and incubate at 37 °C for 24 h.
5.5.2.5.2.2 Using 2 ml of nutrient medium (see 5.4.1 1), wash the bacterial growth from the 24 h culture from the slope, scraping the agar surface if necessary. Carefully decant the suspended growth into a sterile screw-top glass bottle and vortex to suspend all the growth in the medium.
5.5.2.52.3 Add 0,5 ml of the solution obtained in 5.5.2.5.2.2 to 50 m« of nutrient medium (see 5.4.11) and incubate for 2 h at 37 "C.
5.52.524 Verify that the absorbance measured at 620 nm ± 20 nm through cells of optical path 1 cm is between 0,05 and 0,10 (inclusive), representing a bacterial concentration of approximately 10 organisms per millilitre.
532J523 Prepare a tenfold dilution series of the bacteriophage suspension, using, for each step, 9 ml volumes of sterile distilled water and, to 0,1 ml of each of the dilutions obtained, add 0,9 ml of the 2 h £ CO// culture (see 5.5.2.5.2.4).
5.52.52.6 Place the suspensions in a water-bath or incubator at 37 °C for 15 min ± 1 min and then add 5,0 mi of top layer agar, which had been melted and kept in a water-bath at 44 "C to prevent solidification of the agar.
5.5.2.5.2.7 Vortex and immediately spread the mixture over the surface of the bottom layer agar in appropriately labelled Petri dishes, and swirl gently.
5.5.2.5.2.8 Allow the agar to solidify, invert and incubate the Petri dishes at 37 °C for 24 h.
5.5.2.5.2.9 After incubation, examine the plates for plaque formation. Count and record the plaques on each plate that contains between 10 and 100 plaques.
5.5.2.5.2.10 A bacteriophage titre of at least 10^7 per millilitre is recommended.
5.6 Disinfecting efficacy of disinfectants and detergent-disinfectants based on
ciilortiexidine gluconate
5.6.1 Inoculate a bottle of tryptone soy broth (see 5.4.22) from a daily subculture of each of the Staphylococcus aureus (SABS TCC Sta 59) and Pseudomonas aeruginosa (SABS TCC Pse 1 6) (see 5.5.2.1 .2) test organisms.
5.6.2 Incubate the inoculated tryptone soy broth at 37 °C for 24 h and proceed as in 5.5.2.1.3, so standardizing the suspension that it contains at least 10 million (10^) but not more than 100 million (10 )
organisms per millilitre.
5.6.3 Test procedure
5.6.3.1 Using sterile distilled water, prepare a relevant dilution of the disinfectant or detergent- disinfectant (see 7.2(f)).
5.6.3.2 Dispense 9 mi volumes of this sample dilution aseptically into sterile test tubes and place for 10 min in a water-bath maintained at 37 "C. Repeat this procedure but use 9 ml of sterile distilled water as a control.
5.6.3.3 Melt the contents of a sufficient number of bottles (containing 15 ml volumes) of nutrient agar (see 5.4.9), cool to 45 °C and maintain them at this temperature.
5.6.3.4 Using a clean, sterile pipette, dispense 1 m« of the Staphylococcus aureus suspension (see 5.6.2) into one of the tubes of each disinfectant or detergent-disinfectant to be tested and to the control.
Mix well, using a vortex type mixer and maintain the suspension in the water-bath for the duration of the test.
5.6.3.5 Remove a 1 mi volume from each tube after 1 min and add each volume to separate 9 ml volumes of neutralizer broth medium (see 5.4.7.4) and stir well.
5.6.3.6 Prepare a tenfold dilution series of the sample, using for each step 9 ml volumes of sterile distilled water as diluent. Four to five serial dilutions are recommended for the control.
5.6.3.7 Using a clean, sterile pipette, dispense 1 ml of each dilution (see 5.6.3.5 and 5.6.3.6) onto two appropriately labelled, sterile plates (Petri dishes). Add 15 ml! of nutrient agar (see 5.6.3.3) to each of the plates and swirl gently to ensure an even distribution of colonies after incubation. Avoid spilling any of the contents of the plates during this process.
5.6.3.8 Allow the agar to solidify, invert the plates and incubate at 37 °C for 48 h.
5.6.3.9 After incubation, examine the plates for growth. Count and record the colonies on each plate (of the test solution and of the control) that contains between 30 and 300 colonies. If the least diluted sample (with the highest concentration) yields less than 30 colonies, count all the colonies. Ensure that the colonies that have been counted are derived from survivors of the test organisms and not from contamination.
5.6.3.10 Take the dilution factor (DF) for the test sample and for the control as the inverse of the dilution, for example if the sample or the control has been diluted to 1/1 000, take DF as 1 000.
5.6.3.12 Repeat the procedure described in 5.6.3.1 to 5.6.3.1 1 , using Pseudomonas aeruginosa as the test organism.
5.6.4 interpretation of results
Deem the disinfectant or detergent-disinfectant to comply with the requirements of 4.2 if, subject to the following conditions:
a) at a concentration of 2 % of chlorhexidine gluconate (in cases where the supplied disinfectant or detergent-disinfectant contains 5 % of chlorhexidine gluconate or more); or
b) at the prescribed concentration, i.e. in an undiluted form (see 7.2(f)) (in cases where the supplied disinfectant or detergent-disinfectant contains less than 5 % of chlorhexidine gluconate),
the disinfectant or detergent-disinfectant kills at least 99,9 % of each organism tested.
5.7 Disinfecting efficacy of disinfectants and detergent-disinfectants based on
glutaraldeliyde
5.7.1 Survivor count method
The following distinctions are made:
a) disinfectants or detergent-disinfectants intended for general use; and
b) disinfectants intended for use on medical instruments.
5.7.1 .1 Preparation of test organism suspensions
5.7.1.1.1 Disinfectants and detergent-disinfectants based on glutaraldeliyde, intended for general use
Prepare the test organism suspensions as described in 5.5.2.1 and 5.5.2.3 and ensure that each millilitre of growth medium contains:
a) at least 1 00 000 (1 0^5), but not more than 1 million (1 0^) Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa organisms; and
b) 1 00 000 (1 0^5) Aspergillus niger organisms, respectively.
5.7.1.1.2 Disinfectants based on glutaraldeliyde, intended for use on medical instruments
Prepare the test organism suspensions as described in 5.5.2.1 , 5.5.2.3 and 5.5.2.4, and ensure that each
millilitre of growth medium contains:
a) at least 1 million (1 0^7), but not more than 1 00 million (1 0^8) Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa organisms;
b) 1 million (10^7) Aspergillus n/ger organisms; and
c) at least 1 million (1 0^7) Bacillus subtilis var. globigii spores, respectively.
5.7.1.2 Preparation of control and test solutions
5.7.1.2.1 Control solutions
5.7.1.2.1.1 Disinfectants and detergent-disinfectants based on glutaraldehyde, intended for general use
Dispense 8,9 ml of standard hard water No. 1 (see 5.4.1 8.1 ) into each of three sterile glass bottles and to each add 0,1 ml of sterile skimmed milk (see 5.4.20).
5.7.1.2.1.2 bisinfectants based on glutaraldehyde, Intended for use on medical instruments
Dispense 9 ml of sterile distilled water into each of three sterile glass bottles.
5.7.1 .2.2 Test solution
5.7.1.2.2.1 Disinfectants and detergent-disinfectants based on glutaraldehyde, intended for general use
Prepare the prescribed concentration of the test sample as stated on the label (see 7.2(f)), using standard hard water No. 1 (see 5.4.18.1). Dispense 8,9 mfi of the test solution into each of three sterile glass bottles and to each add 0,1 ml of sterile skimmed milk (see 5.4.20). Use the test solution on the day of preparation.
5.7.1.2.2.2 Disinfectants based on glutaraldehyde, intended for use on medical Instruments
Prepare the relevant concentration of the test sample as stated on the label (see 7.2(f)), using sterile distilled water. Dispense 9 m{ of the test solution into each of three sterile glass bottles. Use the test solution on the day of preparation.
5.7.1.3 Temperature adjustment
Label the bottles that contain the control, the test solutions and the test organism suspensions and place them for at least 30 min in a water-bath maintained at 22 °C ± 1 °C.
5.7.1.4 Test procedure for disinfectants and detergent-disinfectants based on glutaraldehyde, intended for general use
NOTE - For the sake of brevity, the procedure for testing the disinfectant solution against one organism is given. For each organism, a control test Is carried out, using the relevant solution (see 5.7.1 .2.1 .1 and 5.7.1 .2.1 .2) instead of the disinfectant solution. Each control runs concurrently with its associated test (with a time lapse of about 30 s between associated actions). Use a stop-watch for this purpose.
5.7.14.1 Melt the contents of a sufficient number of bottles (containing 15 mi volumes) of nutrient agar (see 5.4.9), cool to 45 °C and maintain them at this temperature.
5.7.1.45 Using a clean, sterile pipette, dispense 1 m{ of the Pseudomonas aeruginosa test suspension (see 5.7.1 .1 .1(a)) into the test solution and to the control (without removing the bottles from the water-bath). Remove the bottles from the water-bath, vortex them to mix the contents thoroughly and immediately return them to the water-bath.
5.7.1.4.3 At the end of a 5 min exposure period (actual contact time), transfer 1 mH of the test solution containing the organism to 9 ml of inactivator medium No. 2 (see 5.4.7.2) and mix well into a uniform suspension (first dilution). Within 30 s, repeat this procedure with the control.
5.7.1.4.4 Using a clean, sterile pipette, transfer 1 ml of the inactivator medium containing the organism (see 5.7.1 .4.3) to a Petri dish and then 1 m{ to the first of a series of bottles, each containing 9 ml of sterile distilled water. Mix well into a uniform suspension (second dilution).
5.7.1.4.5 Repeat the procedure in 5.7.1.4.4, but transfer the suspension obtained after the second dilution to the second of the series of bottles. Dilute further until an end dilution of 1 :1 000 of the test sample is obtained.
5.7.1.4.6 Consecutively prepare, as described above, a dilution series using the control.
5.7AA.7 From each dilution of the test sample, transfer 1 m{ to each of two appropriately labelled, sterile plates (Petri dishes). Do the same for each dilution of the control. Add 15 me of nutrient agar (see 5.7.1 .4.1) to each of the plates and swirl gently to ensure an even distribution of colonies after incubation.
Avoid spilling any of the contents of the plates during this process.
5.7.1.4.8 Allow the agar to solidify, invert the plates and incubate them at 37 °C for 48 h.
5.7.1.4.9 After incubation, examine the plates for growth. Count and record the colonies on each plate (of the test solution and of the control) that contains between 30 and 300 colonies. If the least diluted sample (with the highest concentration) yields less than 30 colonies, count all the colonies. Ensure that the colonies that have been counted are derived from survivors of the test organisms and not from contamination.
5.7.1.4.10 Take the dilution factor {DF) for the test sample and for the control as the inverse of the dilution, for example if the sample or the control has been diluted to 1/1 000, take DF as 1 000.
5.7.1.4.11 The percentage kill
5.7.1.4.12), Staphylococcus aureus after 5 min contact time (see 5.7.1.4.13) and Aspergillus mger after 15 min contact time (see 5.7.1.4.14).
5.7,1.4.12 Repeat the procedure described in 5.7.1 .4.1 to 5.7.1 .4.1 1 (inclusive) with the Escherichia coli suspension (see 5.5.2.1).
5.7.14.13 Repeat the procedure described in 5.7.1 .4.1 to 5.7.1 .4.1 1 (inclusive) with the Staphylococcus aureus suspension (see 5.5.2.1).
5.7.14.14 Repeat the procedure descritDed in 5.7.1 .4.1 to 5.7.1 .4.1 1 (inclusive) with the Aspergillus niger
test organism suspension (see 5.5.2.3), but
a) use an exposure period (actual contact time) of 1 5 min,
b) add malt extract agar (see 5.4.8) to the suspensions on the plates, and
c) incubate lor 7 d at 25 °C.
5.7.1.5 Test procedure for disinfectants based on glutaraidehyde. intended for use on medical instruments
5.7.1.5.1 Pseudomonas aeruginosa and Aspergillus nIger
Carry out the procedure in 5.7.1 .4.1 to 5.7.1 .4.1 1 (inclusive), using the Pseudomonas aeruginosa test suspension (see 5.7.1 .1 .1(a)) and the Aspergillus nigenesX suspension (see 5.5.2.3).
5.7.1.5.2 Bacillus subtilis var. globigii
Carry out the procedure in 5.7.1 .4.1 to 5.7.1 .4.1 1 (inclusive), using the Bacillus subtilis var. globigii test suspension (see 5.5.2.4), but:
a) use an exposure period (actual contact time) of 4 h; and
b) incubate at 37 °C for 48 h.
5.7.1.6 Interpretation of results
Deem the samples to comply with the requirements of 4.2 if. for each organism tested, a result of at least a 99,99 % kill was obtained.
5.7.2 Sporicidal activity (Kelsey-Sykes test modified)
5.7.2.1 Medium
Use the inactivator medium No. 2 described in 5.4.7.2.
5.7.2.2 Test organism suspension
Use the spore suspension of B. subtilis var. globigii (see 5.5.2.4).
5 7.2.3 Preparation of test solution
Prepare the prescribed concentration of the test solution as stated on the label (see 7.2(f)). using sterile distilled water. Use the test solution on the day of preparation.
5.7.2.4 Test procedure
5.7.24.1 Dispense 3 ml of the test solution into a sterile glass bottle.
5 7.24.2 Label the bottles containing the test solution and the test organism suspensions and place them for at least 30 min in a water-bath maintained at 22 °C ± 1°C.
5.7.24.3 Then, without removing the bottle containing the test solution from the water-bath and using a clean, sterile pipette, add 1 m{ of the test organism suspension to the test sample. Simultaneously start a stop-watch. Remove the bottle from the water-bath, mix the contents well and immediately return it to the water-bath.
5.7.2.4.4 After exactly 4 h, transfer 0,02 ml of the suspension obtained in 5.7.2.4.3 to each of five tubes containing 10 ml of inactivator medium No. 2 (see 5.4.7.2).
5.7.2.4.5 Incubate the inoculated tubes of inactivator medium at 30 °C for 48 h.
5.7.2.4.6 After incubation, examine the tubes of inactivator medium for growth and record the results.
5.7.2.5 Interpretation of results
Deem the sample to comply with the requirements of 4.2 if no growth of the test organism is detectable in any of the five tubes of inactivator medium.
5.7.3 Kelsey-Sykes test
5.7.3.1 Procedure
Follow the procedure in 5.8, but use only Pseudomonas aeruginosa as the test organism.
5.7.3.2 Interpretation of results
Deem the sample to comply with the requirements of 4.2 if the initial concentration of disinfectant (concentration B) (see 5.8.2.6(a)) shows no growth of the test organism in at least two of the five tubes of reculture medium in sets inoculated at:
a) the eighth minute after the addition of the initial inoculum; and
b) the eighteenth minute after the addition of the initial inoculum.
An example of a series of test results and their interpretation is given in table 2.
5.8 Kelsey-Sykes test for detergent-disinfectants based on plienolics
5.8.1 Minimum inhibitory concentration test
5.8.1.1 Preparation of the test suspensions
Prepare a 1 :10 dilution in Wright and Mundy medium (see 5.4.23) of a freshly grown subculture of each of the test organisms (see 5.5.2.1).
NOTE - Before diluting a Pseudomonas aeruginosa culture, filter it through a coarse filter paper.
5.8.1 .2 Preparation of the test sample dilutions
5.8.1.2.1 To 5 ml of test sample in a glass bottle of capacity 30 ml (see 5.2.1 (a)), add 5 ml of Wright and Mundy medium (see 5.4.23). Mix well and transfer 5 mi of this dilution of test sample to a further 5 mi of Wright and Mundy medium. Repeat the procedure until 10 doubling dilutions of the test sample (from 1 :2 to 1 :1 024) have been prepared. Discard 5 mi of the last dilution (so that each bottle will contain 5 m{ of a dilution of the test sample).
5.6.122 Repeat 5.8.1 .2.1 until three sets of 10 doubling dilutions of the test sample have been prepared.
5.8.1.3 Test procedure
5.8.1.3.1 To each of the 10 dilutions of the test sample (see 5.8.1.2.2), add 0,02 mi of the
Staphylococcus aureus test suspension (see 5.8.1 .1). Incubate the inoculated bottles at 30 °C for 72 h.
Examine the bottles for growth. The minimum Inhibitory concentration is the highest dilution (minimum concentration) not showing growth.
5.8.1.3.2 Repeat the procedure given In 5.8.1.3.1 but use, successively, the Escherichia coll and Pseudomonas aeruginosa (see 5.8.1 .1) test suspensions.
5.8.1.4 Interpretation of results
Determine which of the three test organisms is most resistant to the test sample, i.e. the organism for which minimum concentration Is the highest. Use this organism for the remainder of the test (see 5.8.2).
5.8.2 Remainder of test
5.8.2.1 Selection of the test organism for the determination using the minimum inhibitory concentration test (see 5.8.1 ). determine which of the test organisms is the most resistant, and use It as the test organism for the determination.
5.8.2.2 Preparation of culture for test organism suspensions
On the day before the test is due to be carried out, inoculate a tube containing 10 ml of Wright and Mundy medium (see 5.4.23) from a daily subculture of the appropriate test organism (see 5.5.2.1 ) and incubate the inoculated medium at 37 °C for 24 h.
5.8.2.3 Preparation of test organism suspension for the test under "clean" conditions
5.8.2.3. 1 After incubation (see 5.8.2.2). centrifuge the culture of the test organism for 15 min at a resultant centrifugal of 6 kN/kg. using a sterile Pasteur pipette, remove and discard the supernatant liquid and resuspend the organism In 1 mi! of hard water prepared for use in this test (see 5.4.1 8.3).
5.8.2.3.2 Transfer this suspension to a sterile glass bottle of capacity 30 ml (see 5.2.1 (a)).
5.8.2.3.3 Add a few sterile glass beads and shake for 1 min.
5.8.2.4 Preparation of test organism suspension for the test under "dirty" conditions
Obtain a suspension that contains 5 % (by mass) of dry yeast by adding 6 ml of the culture of the test organism (see 5.8.2.3) to 4 md of 5 % (by mass) of the dry yeast suspension (see 5.4.24.4) contained in a sterile glass bottle of capacity 30ml (see 5.2.1 (a)). Add a few sterile glass beads to the mixture and vortex for 1 min.
5.8.2.5 Estimation of the number of viable organisms in the test organism suspension
So standardize the suspension by using a spectrophotometer in conjunction with a standard curve, a haemocytometer, Petroff-Hausser counting chamber or any other suitable means, that it contains at least 100 million (10^6) but not more that 10^10 organisms per milliliter. Use the suspension with three hours of prepearation.
5.8.2.6 Preparation of test solutions
Using the hard water prepared for this test (see 5.4.18.3) and glass bottles of capacity 30 ml (see 5.2.1 (a)), prepare three different concentrations A. B and C of the test sample that are such that.
a) concentration B is that which is expected or claimed to pass the test;
b) concentration A is half of concentration B ; and
c) concentration C is one-and-a-half-times concentration B.
For example, if a test sample is expected to pass the test at a concentration of 1 %, concentration A is a 0,5 % concentration, B is a 1 % concentration and C is a 1 ,5 % concentration.
5.8.2.7 Test procedure under "clean" conditions
5.8.2.7.1 Dispense 3 ml of each concentration of the test sample (see 5.8.2.6) into glass bottles of capacity 30 ml (see 5.2.1(a)), and label these bottles A, B and C, as relevant.
5.8.2.7.2 Place these bottles and the bottle containing the test organism suspension (see 5.8.2.3) for at least 30 min in a water-bath maintained at 22 °C ± 1 °C. To maintain the reproducibility of the test, adhere strictly to the temperature stated.
5.8.2.7.3 Then, without removing the bottle containing test concentration A from the water-bath, add 1 ml of the test suspension (see 5.8.2.3) and at the same time start a stop-watch (zero time). Remove the bottle from the water bath, mix well and immediately return it to the water-bath.
5.8.2.7.4 One minute after zero time, add, in the same way, 1 ml of the test suspension to the bottle containing test concentration B.
5.8.2.7.5 Five minutes after zero time, add, in the same way, 1 ml. of the test suspension to the bottle containing test concentration C.
5.8.2.7.6 Eight minutes after zero time, transfer 0,02 m{ from the bottle containing test concentration A to each of five tubes of reculture medium (see 5.4.1 4) each of which has been labelled A1 .
5.8.2.7.7 Ten minutes after zero time, add, in the same way as described in 5.8.2.7.3 above, a further 1 ml of test suspension (see 5.8.2.3) to the bottle containing test concentration A.
5.8.2.7.8 Eighteen minutes after zero time, transfer 0,02 me from the bottle containing test concentration A to each of five tubes of reculture medium each of which has been labelled A2.
5.8.2.7.9 Concurrently with 5.8.2.7.1 to 5.8.2.7.8. treat the test concentrations B and C in the same way, but base the time intervals on the times at which the first additions of the test suspensions were made, and label the sets of tubes of reculture medium B1 and B2, and C1 and C2 (respectively). Thus in the case of test concentration B, addition and transference will be made one minute later than the times given for test concentration A (i.e at 9 min , 11 min and 19 min after zero time).
Note – In order to obviate errors in transference, a copy of the test timetable (see table 2) should be used during each test. Each step of the test should be ticked off on the timetable as it is carried out.
5.8.2.7.10 Incubate all the inoculated tubes of reculture medium at 30 °C for 48 h.
5.82.7.11 After incubation, examine the tubes of reculture medium for growth and record the results as
shown in the example given in table 3.
5.8.2.7.12 If test concentration B passes the test (see 5.8.2.8), repeat steps 5.8.2.7.1 to 5.8.2.7.1 1 on two subsequent days.
5.8.2.8 Test procedure under "dirty" conditions
carry out the procedure described in 5.8.2.7 but use the test organism suspension described in 5.8.2.4.
5.9 Disinfecting efficacy of coal-tar type disinfectant liquids (black and white)
— Rideai-Wailcer coefficient test
5.9.1 Preparation of test organism suspension
Mix thoroughly a culture of Salmonella typhi (see 5.5.2.2) in nutrient broth No. 2 (double strength) (see 5.4.10) that has been incubated for 24 h at 37 °C and, before use, place the tube for 30 min ± 5 min in the water-bath (see 5.3.2) maintained at 22 °C.
5.9.2 Preparation of control and test solution
5.9.2.1 Standard phenol control solution
From the 50 g/l of standard phenol solution (see 5.4.1 9), make five phenol control dilutions that contain 1 g of pure phenol in each of 95 mi, 100 ml, 105 mi 1 10 ml and 1 15 m£ of solution.
NOTE - These dilutions may be stored in the dark for not more than one week before use.
5.9.2.2 Stock solution of the test sample (1 :100)
5.9.2.2.1 Immediately before withdrawing any portion for testing, thoroughly mix a composite sample of the disinfectant to be tested, ensuring that air is not beaten into or shaken into the sample.
5.9.2.2.2 Withdraw the test portion from the middle of the sample by means of a 5 ml capacity pipette (see 5.2.3). Fill the pipette to just above the mark, wipe it clean on the outside with sterile cotton wool and then adjust the contents to the mark. Allow the contents of the pipette to discharge below the surface of about 480 ml of water at a temperature of 1 8 °C, contained in a measuring cylinder.
5.9.2.2.3 Rinse the pipette out three times (or more in the case of viscous liquids) by drawing up and returning some of the dilution.
5.9.2.2.4 Make the solution up to 500 ml with water, stopper the cylinder and thoroughly mix the contents by inverting, with a corkscrew motion, 50 times.
5.9.2.3 Test solutions
From the stock solution (see 5.9.2.2), prepare five suitable test solutions (see tables 4 and 6) based on the nominal RW coefficient stated on the label (see 7.2(m)). Place 5 me of each of the five chosen solutions in sterile culture tubes (see 5.2.2(a)). Mark and place these tubes in sequence in a rack in the water-bath (see 5.3.2), with the strongest disinfectant solution on the left.
5.9.2.4 Phenol solutions
Prepare five culture tubes (see 5.2.2(a)), each containing 5 m{ of a different phenol control solution (see 5.9.2.1), and mark and arrange them in the same way in the water-bath as the solutions of the disinfectant (see 5.9.2.3).
5.9.2.5 Culture medium
5.9.2.5.1 Mix thoroughly a culture of Salmonella typhi (see 5.5.2.2) in Nutrient broth No. 2 (double strength (see 5.4.10) that has been incubated at 37 °C for 24 h and, before use, place the tube for 30 min ± 5 min in a water-bath maintained at 22 "C.
5.9.2.5.2 Place two sets of 15 tubes (marked sequentially "1 " to "30"), each containing 5 mi of Nutrient broth No. 2 (double strength) (see 5.4.10), in the water-bath.
5.9.3 Test procedure
5.9.3.1 Starting exactly at zero time (use the stop-watch (see 5.3.4) for timing all operations), add 0,2 mlof the test organism suspension (see 5.9.1 ) from the droplet pipette (see 5.2.1 0) to the leftmost test tube containing the disinfectant solution (see 5.9.2.3). Ensure that all of the culture added is pipetted straight into the disinfectant solution and not onto the wall of the test tube. Shake the tube; 30 s after this, inoculate the next tube to the right with 0,2 ml of culture in a similar manner. Inoculate each successive tube, at intervals of 30 s, until the fifth tube has been inoculated.
5.9.3.2 Thirty seconds after this last addition, i.e. 2,5 min after zero time, withdraw a representative loopful (see 5.2.9) of the well-shaken contents of the tube on the extreme left and add this to the tube marked "1" and containing 5 ml of the nutrient broth No. 2 (double strength) (see 5.4.10). Immediately after inoculation, shake the tube containing the inoculated broth. Thirty seconds after this loopful has
been withdrawn, transfer, in the same way. a loopful of the contents of the second test tube to the tube of broth marked "2". Repeat this procedure at intervals of 30 s, working from left to right, until all five test tubes have been so treated.
5 9.3.3 Starting again in each case with the left hand tube, perform two further cycles of withdrawals until three sets of cultures (15 tubes) have been made, i.e. at 2.5 min, 5 min and 7,5 mm intervals, respectively, after exposure.
5.9.34 Repeat the procedure given in 5.9.3.1 to 5 - Reference of the measure
- Regulation 5 and 6
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 3808.94 -- Disinfectants Yes Disinfectants and detergent-disinfectants intended for use on inanimate surfaces. - Description
- Disinfectants and detergent-disinfectants intended for use on inanimate surfaces.
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 9401.20: - Seats of a kind used for motor vehicles
The measure came into effect on 20 August 2003
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 20 August 2003
- Publication where the measure is specified
- Government Notice No. R. 862 (Government Gazette 25082) Of 20 June 2003
- Regulation where the measure is specified
- Compulsory Specification For Child Restraints For Use In Motor Vehicles
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This specification applies to child restraint that are suitable for installation in power driven vehicles that have three or more wheels.
- Coded list of objectives
- X: For purposes n.e.s.
- Description of the measure
- 6.1.4 Dynamic test
6.1.4.1 General
6.1.4.1.1 Child restraints of the universal, the restricted and the semi-universal category shall be tested on the test trolley using the test seat described in F.3 of this specification, and in accordance with 7.1.3.1.
6.1 .4.1 .2 Child restraints of the specific vehicle category shall be tested with each vehicle model for which the child restraint is intended. The test authority responsible for conducting the tests may reduce the number of vehicle models tested if the models do not differ much In respect of the features mentioned in 6.1.4.1.3(c).
6.1 .4.1 .3 Child restraints of the specific vehicle category may be tested in one of the following ways:
a) using a vehicle body shell on the test trolley, as given in 7,1.3.2; or
b) using a complete vehicle, as given in 7.1.3.3; or
c) using a sufficient number of parts of the vehicle body shell to be representative of the vehicle structure and impact surfaces.
If the child restraint is intended for use on the rear seat, the test areas shall include the back of the front seat, the rear seat, the floor pan, the B and C pillars, and the roof.
If the child restraint Is intended for use on the front seat, the testareas shall include the dashboard, the A pillars, the windscreen, any levers or knobs installed in the floor or on a console, the front seat, the floor pan and the roof.
If the child restraint is intended for use in combination with the adult safety belt{s), the appropriate adult belt(s) shall be included in the test.
The test authority responsible for conducting the tests may permit parts to be excluded from the test if they are found to be superfluous. The tests shall be conducted in accordance with 7.1.3.2.
6.1 .4.1 .4 If a child restraint system of the specific vehicle category is installed in the area behind the rearmost forward-facing adult seat position (for example, the luggage area), one test with the largest dummy on a complete vehicle, as given in 7. 1 .3.3, shall be performed. The tests given in 7. 1.3.2 may be conducted if required by the manufacturer.
6.1 .4.1 .5 In the case of a special-needs restraint every dynamic test specified in this specification for each mass group shall be performed twice: first, using the primary means of the restraint and, second, with all restraining devices in use. In these tests, special attention shall be given to the requirements given in 5.2.3 and 5.2.4.
6.1 .4.1 .6 In the case of a restraint of the non-integral class the safety belt used shall be the standard belt and its anchorage brackets shall be as given in annex B of this specification. This requirement does not apply to child restraints of the specific vehicle category where the safety belt of the vehicle shall be used.
6.1 .4.1 .7 The dynamic tests shall be conducted on child restraints that have not previously been under load.
6.1 ,4,1 .8 During the dynamic tests, no part of the child restraint that actually helps to keep the child in position shall break, no buckles or locking system or displacement system shall release, and the standard safety belt used to install the child restraint shall not become disengaged from any guide or locking device utilized in the tests.
6.1.4.2 Chest acceleration
6.1.4.2.1 The resultant chest acceleration during the dynamic tests shall not exceed 540 m/s^2, except during periods whose sum does not exceed 3 ms.
6,1.4.2.2 The vertical component of the acceleration from the abdomen towards the head shall not exceed 295 m/s^2. except during periods whose sum does not exceed 3 ms.
NOTE Chest acceleration limits do not apply when the newborn test manikin is used.
6.1.4.3 Abdominal penetration
During the verification described in G.4.5.3 of this specification, there shall be no visible sign that any part of the child restraint has penetrated the modelling clay in the abdomen,
7 Description of tests
7 A Tests of the assembled restraint
7.1.1 Corrosion
7.1.1.1 Position the metal items of the child restraint in a test chamber, in accordance with annex H of this specification. In the case of a child restraint that incorporates a retractor, unwind the strap to its full length minus 100 mm ± 3 mm. Except for short interruptions that might be necessary, for example to check and replenish the salt solution, the exposure test shall proceed continuously for a
period of 50h.
7.1.1.2 On completion of the exposure test, gently wash, or dip. the metal items of the child restraint in clean running water at a temperature not higher than 38 ̊ C, to remove any salt deposit that might have formed. Allow to dry at a room temperature of 18 ̊ C to 25 ͦ C for 24 h and inspect in accordance with 6.1.1.2,
7.1.2 Overturning
7.1 .2.1 Place the test manikin in the restraint installed in accordance with this specification and taking into account the manufacturer's Instructions and with the standard slack as specified in 7.1 .3.6.
7.1.2.2 Fasten the restraint to the test seat or vehicle seat. Rotate the entire seat around a horizontal axis contained in the median longitudinal plane of the seat through an angle of 360" at a speed of 2 7s to 57s. For the purposes of this test, a restraint intended for use in specific vehicles may be attached to the test seat described in annex F of this specification.
7.1.2.3 Repeat the test while rotating the seat in the reverse direction after, if necessary, the test manikin has been replaced in its initial position. With the rotational axis In the horizontal plane and at 90ͦ to that of tile two earlier tests, repeat the procedure in the two directions of rotation.
7.1.2.4 Do these tests while using both tile smallest and the largest appropriate manikin of the group(s) for which the restraining device is intended.
7.1.3 Dynamic tests
7.1 .3.1 Tests using the trolley and the test seat
7.1.3.1.1 Forward facing
7.1.3.1.1,1 The trolley and test seat used in this dynamic test shall meet the requirements given in annex F of this specification, and the dynamic crash test installation procedure shall be in accordance with annex I.
7.1.3.1.1.2 The trolley shall remain horizontal throughout deceleration.
7.1.3.1.1.3 Achieve the deceleration of the trolley by using the apparatus given in annex F of this specification, or any other device that gives equivalent results. This apparatus shall be capable of the performance specified in 7.1.3.4 and in annex J of this specification.
7.1.3.1.1.4 Make the following measurements and inspections:
a) the trolley speed, immediately before impact;
b) the stopping distance;
c) the displacement of the test manikin's head in the vertical and horizontal planes for group I, group II and group III restraints and, for group and group 0+ restraints, the displacement of the manikin (other than that of Its limbs);
d) the chest acceleration in three mutually perpendicular directions, except in the case of the newborn test manikin; and
e) any visible signs of penetration of the modelling clay in the abdomen (see 6.1.4.3), except in the case of the newborn test manikin.
7.1.3.1.1.5 Film the tests at frequency at least 500 frames per second.
7.1.3.1.1.6 After impact visually inspect the child restraint without opening the buckle, to determine whether there has been any failure or breakage.
7.1.3.1,2 Rearward facing
7.1 .3.1 .2.1 Rotate the test seat through an angle of 1 80^ in accordance with the requirements of the rear impact test
7.1.3.1.2.2 When a rearward-facing child restraint intended for use in the front seating position is being tested, the vehicle facia shall be represented by a rigid bar so attached to the trolley that all the energy absorption takes place in the child restraint.
7.1.3.1.2.3 The deceleration conditions shall satisfy the requirements of 7.1.3.4.
7.1.3.1.2.4 Make the measurements as given in 7.1.3.11.4.
7.1,3.1.2.5 Film the tests at least 500 frames per second.
7.1,3.1.2.6 After impact visually inspect the child restraint without opening the buckle, to determine whether there has been any failure or breakage.
7.1.3.2 Test using the trolley and the vehicle body shell
7.1.3.2.1 Forward facing
7.1 .3.2.1 .1 The method used to secure the vehicle during the test shall not be such that the anchorages of the vehicle seats, adult safety belts and any additional anchorages required to secure the child restraint will be strengthened, or such that the normal deformation of the structure will be lessened.
No part of the vehicle shall be present which, by limiting the movement of the test manikin, would reduce the load imposed on the child restraint during the test. The eliminated parts of the structure may be replaced by parts of equivalent strength, provided that they do not hinder the movement of the test manikin.
7.1 .3.2.1 .2 A securing device shall be regarded as satisfactory if it produces no effect on an area that extends over the entire width of the structure and if the vehicle or structure is blocked or fixed in front at a distance of not less than 500 mm from the anchorage of the restraint system. At the rear, the structure shall be secured at a sufficient distance behind the anchorages to ensure that the requirements of 7.1.3.2.1.1 are satisfied.
7.1.3.2.1 .3 The vehicle seat and child restraint shall t)e fitted and shall be placed in a position that has been selected by the test authority to give the most adverse conditions in respect of strength, compatible with installing the test manikin in the vehicle. The position of the vehicle seat back and the child restraint shall be stated in the report. The vehicle seat back, if adjustable for inclination, shall be locked as specified by the manufacturer or, in the absence of any specification, at an actual seat back angle as near as possible to 25 ͦ
7.1.3.2.1.4 Unless the instructions for fitting and use require otherwise, the front seat shall be placed in the most forward position normally used in the case of child restraints that are intended for use in the front seating position, and in the rearmost position normally used in the case of child restraints that are intended for use in the rear seating position.
7.1.3.2.1.5 The deceleration conditions shall satisfy the requirements of 7.1.3.4. The test seat shall be the seat of the actual vehicle.
7.1 .3.2.1 .6 Make the following measurements and inspections:
a) the trolley speed, immediately before impact;
b) the stopping distance;
c) any contact of the test manikin's head (in the case of group 0, not taking the manikin's limbs into account) with the interior of the vehicle body shell;
d) the chest deceleration in three mutually perpendicular directions, except in the case of the newborn test manikin; and
e) any visible signs of penetration of the modelling clay in the abdomen (see 6.1 .4.3). except in the case of the newborn test manikin.
7.1.3.2.1.7 Film the tests at least 500 frames per second.
7.1 .3.2.1.8 After impact, visually inspect the child restraint, without opening the buckle, to determine whether there has been any failure or breakage.
7.1.3.2.2 Rearward facing
7.1.3.2.2.1 For rear impact tests, rotate the vehicle body shell through an angle of 1 80' on the test trolley.
7.1.3.2.2.2 In all other respects, the requirements for frontal impact apply.
7.1.3.3 Test using the complete vehicle
7.1 .3.3.1 The deceleration conditions shall satisfy the requirements of 7.1.3.4.
7.1.3.3.2 For frontal impact tests, use the procedure set out in annex K of this specification.
7.1.3.3.3 For rear impact tests, use the procedure set out in annex L of this specification.
7.1.3.3.4 Make the following measurements and inspections:
a) the speed of the vehicle or the impactor, immediately before impact;
b) any contact of the manikin's head (in the case of group 0, not taking the manikin's limbs into account) with the interior of the vehicle;
c) the chest acceleration in three mutually perpendicular directions, except in the case of the newborn test manikin; and
d) any visible signs of penetration of the modelling clay in the abdomen (see 6.1 .4.3), except in the case of the newborn test manikin.
7.1 .3.3.5 Film the tests at at least 500 frames per second:
7.1.3.3.6 If the front seats are adjustable for inclination, lock the seats as specified by the manufacturer or, in the absence of any specification, at an actual seat back angle as near as possible to 25ͦ. .
7.1 .3.3.7 After impact, visually inspect the child restraint, without opening the buckle, to determine whether there has been any failure or breakage.
7.1.3.4 Conditions for dynamic tests
The conditions for dynamic tests are summarized in table 1.
7.1.3.5 Child restraints that require the use of additional anchorages
7.1.3.5.1 In the case of semi-universal child restraints that require the use of additional anchorages, the frontal impact test, in accordance with 7.1.3.4. shall be carried out as given in 7.1.3.5.2 to 7.1.3.5.5.
7.1.3.5.2 In the case of restraints with short upper attachment straps, for example restraints intended to be attached to the rear parcel shelf, the upper anchorage configuration on the test trolley shall be as given in F.5 of this specification.
7.1.3.5.3 In the case of restraints with long upper attachment straps, for example restraints intended for use where there is no rigid parcel shelf and where the upper anchorage straps are attached to the vehicle floor, the anchorages on the test trolley shall be as given in F.5 of this specification.
7.1.3.5.4 In the case of restraints intended for use in both configurations, the test that uses the anchorage configurations given in 7.1 .3.5.2 and 7.1.3.5.3 shall be carried out, except that, in the case of the test that uses the anchorage configurations given in 7.1 .3.5.3, only the heavier manikin shall be used.
7.1.3.5.5 In the case of rearward-facing restraints the lower anchorage configuration on the test trolley shall be as given in F.S of this specification.
7.1.3.6 Test manikins
7.1.3.6.1 General
The test manikins used to test the child restraint shall comply with annex G of this specification.
7.1.3.6.2 installation of the test manikin
7.1.3.6.2.1 Frontal impact with forward-facing restraints and rear Impact with rearward-facing restraints
Place the test manikin so that there is a gap between the front of the manikin and the restraint.
7.1.3.6.2.2 Forward Impact with rearward-facing restraints
Place the test manikin so that there is a gap between the rear of the manikin and the restraint.
7.1.3.6.2.3 Carry-cots
Place the test manikin in a straight horizontal position as close as possible to the centre-line of the carry-cot.
7.1.3.6.2.4 Child restraint with a separately anchored chair
7.1.3.6.2.4.1 Place the test manikin in the vehicle seat or test seat. Place a board 25 mm thick and 60 mm wide between the back of the test manikin and the backrest of the vehicle seat or test seat. The board should follow as closely as possible the curvature of the chair and its lower end should be at the height of the manikin's hip joint Adjust the belt in accordance with the manufacturer's Instructions, but to a tension of 250 N ± 25 N greater than the adjusting force» with the deflection angle of the strap at the adjuster measuring 45ͦ ± 5̊ , or alternatively, at the angle prescribed by the manufacturer. Remove the board.
7.1.3.6.2.4.2 The longitudinal plane that passes through the centre-line of the manikin shall be set midway between the two lower belt anchorages; however, note shall also be taken of 7.1.3.2.1.3. In the case of booster cushions to be tested with the test manikin that represents a 1 0-year-old child, the longitudinal plane that passes through the centre-line of the manikin shall be positioned to the left or right of the point midway between the two lower belt anchorages.
7.1 .3.6.2.4.3 In the case of restraints that require the use of a standard belt, the shoulder strap may be positioned on the test manikin before the dynamic test by means of a lightweight masking tape of sufficient width and length, in the case of rearward-facing devices, the head of the manikin may be held against the backrest of the restraint system by means of a lightweight masking tape of sufficient width and length.
7.1.3.7 Category of test manikin
7.1 .3,7.1 The following categories of test manikin shall be used when testing restraints of the various mass groups:
a) group restraint: test using a newborn manikin and a manikin of mass 9 kg;
b) group 0+ restraint: test using a newborn manikin and a manikin of mass 11 kg;
c) group I restraint: test using manikins of mass 9 kg and 15 kg respectively;
d) group It restraint: test using manikins of mass 15 kg and 22 kg respectively; and
e) group Hi restraint: test using manikins of mass 22 kg and 32 kg respectively.
7.1.3.7.2 If the child restraint system is suitable for two or more mass groups, the tests shall be carried out while using the lightest and heaviest manikins specified above for all the groups concerned. However, if the configuration of the restraint alters considerably from one group to the next, for instance when the configuration of the harness or the length of the harness is changed, the test authority may. if it deems it advisable, add a test with a manikin of intermediate mass.
7.1 .3.7.3 If the child restraint system is designed for two or more children, one test shall be carried out with the heaviest manikins occupying all the seat positions. A second test shall be carried out with the lightest and the heaviest manikins specified above. The test authority may, if it deems it advisable, add a third test with any combination of manikins or empty seat positions.
7.2 Tests of individual components
7.2.1 Buckle
7.2.1 .1 Opening test under load
7.2.1.1.1 Use a child restraint that has already been subjected to the dynamic test specified in 7.1,3 for this test
7.2.1.1.2 Remove the child restraint from the test trolley or from the vehicle, without opening the buckle. Apply a tension of 200 N ± 2 N to the buckle, if the buckle is attached to a rigid part, apply a force that reproduces the angle formed between the buckle and that rigid part during the dynamic test.
7.2.1.1.3 Apply a load at a speed of 400 mm/min ± 20 mm/min to the geometric centre of the buckle-release button, along a fixed axis running parallel to the initial direction of motion of the button. The geometric centre applies to that part of the surface of the buckle to which the release pressure is to be applied. Secure the buckle against a rigid support during the application of the opening force.
7.2.1 .1 .4 Apply the buckle opening force, using a dynamometer or similar device in the manner and direction of normal use. The contact end shall be a polished metal hemisphere of radius 2,5 mm ±0,1 mm.
7.2.1.1.5 Measure the buckle opening force and note any failure.
7.2.1 .2 Opening test under zero load
7.2.1.2.1 Mount and position under a zero load condition a buckle assembly that has not previously been subjected to a load.
7.2.1.2.2 Use the method of measuring the buckle opening force as given in 7.2.1.13 and 7.2.1.1.4.
7.2.1.2.3 Measure the buckle opening force.
7.2.1.3 Strength test
7.2.1.3.1 Use two samples for the strength test. All adjusters, except adjusters mounted direct on a child restraint, are included in the test
7.2.1.3.2 Annex M shows a typical device for a buckle strength test The buckle is placed on the upper round plate A within the relief. All the adjacent straps have a length of at least 250 mm and are arranged hanging down from the upper plate respective to their position at the buckle. The free strap ends are then wound round the lower round plate B until they come out at the plate's inner opening. All the straps shall be vertical between A and B. The round clamping plate C is then clamped lightly against the lower face of B, still allowing a certain strap movement between them. With a small force at the tensile machine, the straps are tensioned and pulled between B and C until all the straps are loaded respective to their arrangement The buckle stays free from plate A or any parts at A during this operation and the test itself. B and C are then clamped firmly together and the tensile force is increased at a traverse speed of 100 mm/min ± 20 mm/min until the required values are reached.
7.2.2 Adjusting device
7.2.2.1 Ease of adjustment
7.2.2.1.1 When testing a manual adjusting device, steadily draw the strap through the adjusting device, having regard for the normal conditions of use, at a rate of approximately 1 00 mm/s. After the first 25 mm of strap movement, measure, to the nearest newton, the maximum force.
7.2.2.1.2 Carry out the test in both directions of strap travel through the device, white the strap is subjected ten times to the full travel cycle, before making the measurement.
7.2.2.2 Microslip test (see annex N, figure N.3)
7.2.2.2.1 Keep the components and adjusting devices to be subjected to the microslip test in an atmosphere at a temperature of 20 ̊ C ± 5 ͦ C and a relative humidity of (65 ± 5) % for a minimum of 24 h before testing. Carry out the test at a temperature of between 15 **C and 30 °C.
7.2.2.2.2 Arrange the free end of the strap in the configuration in which it is when the adjusting device is in use in the vehicle. The free end of the strap shall not be attached to any other part.
7 J2.2.2.3 Place the adjusting device on a vertical piece of strap, one end of which bears a 50 N load (guided so that the load is prevented from swinging and the strap from twisting). Mount the free end of the strap vertically upwards or downwards from the adjusting device, as it is in the vehicle. Pass the other end over a deflector roller, with its horizontal axis parallel to the plane of the section of strap that
supports the load, and the section that passes over the roller being horizontal.
7.2.2.2.4 Arrange the adjusting device under test so that its centre, in the highest position to which it can be raised, is 300 mm ± 20 mm from a support table and the load of 50 N Is 1 00 mm 1 20 mm from the said support table.
7.2.2.2.5 Complete 20 pretest cycles and then complete 1 000 cycles at a frequency of 0.5 cycles per second, the total amplitude being 300 mm ± 20 mm, eras specified in 7.2.4.2.6.2. Apply the 50 N load only during the time that corresponds to a shift of 100 mm ± 20 mm for each half period. Measure the microslip from the position at the end of the 20 pretest cycles.
7.2.3 Retractor
7.2.3.1 Retracting force
Measure the retracting forces with the child restraint fitted with a test manikin as for the dynamic test given in 7. 1.3. Measure the strap tension at the point of contact with (but just dear of) the manikin, while the strap is retracted at an approximate rate of 0,6 m/min.
7.2.3.2 Durability of the retractor mechanism
Withdraw the strap and allow it to retract for the required number of cycles, at a rate of not more than 30 cycles per minute. In the case of emergency-locking retractors, introduce, at each fifth cycle, a jolt to lock the retractor The jolts occur in equal numbers at each of live different extractions, namely at 90 %, 80 %, 75 %, 70 % and 65 % of the total length of the strap on the retractor. However, where the length of the strap exceeds 900 mm, the above percentages are related to the final 900 mm of strap that can be withdrawn from the retractor.
7.2.3.3 Locking of the emergency-locking retractors
7.2.3.3.1 Test the retractor once for locking, when the strap has been unwound to its full length minus 300 mm ±3 mm.
7.2.3.3.2 In the case of a retractor that is actuated by strap movement, the extraction shall be in the direction in which it normally occurs when the retractor is installed In a vehicle.
7.2.3.3.3 When retractors are being tested for sensitivity to vehicle acceleration, they shall be tested at the above extraction length in both directions along two mutually perpendicular axes that are horizontal if the retractors are to be installed in a vehicle as specified by the child restraint manufacturer. When this position is not specified, the test authority shall consult the child restraint manufacturer. One of these test directions shall be selected by the test authority to give the most
adverse conditions with respect to actuation of the locking mechanism,
7.2.3.3.4 The design of the apparatus used shall be such that the required acceleration is given at an average rate of increase of acceleration of at least 245 m/s^2.
7.2.3.3.5 For testing for compliance with the requirements of 6.2.3.2.1.3 and 6.2.3.2.1 .4, the retractor shall be mounted on a horizontal table and the table tilted at a speed not exceeding 2ͦ^ /s until locking has occurred. The test shall be repeated with tilting In other directions, to ensure that the requirements are satisfied.
7.2.3.4 Corrosion test
Carry out the corrosion test as described in 7.1,1.
7.2.3.5 Dust-resistance test
7.2.3.5.1 Position the retractor in a test chamber, as shown in annex Q of this specification. Mount the retractor in an orientation similar to that in which it is mounted in the vehicle. The test chamber shall contain dust as specified in 7.2.3.5,2. Extract from the retractor, and keep extracted, a length of 550 mm of the strap, except that it shall be subjected to ten complete cycles of retraction and withdrawal within 1 min or 2 min after each agitation of the dust For a period of 5 h, agitate the dust every 20 min for 5 s by compressed air that is free of oil and moisture, is at a gauge pressure of 550 kPa ± 50 kPa and enters through an orifice of diameter 1 ,5 mm ± 0,1 mm.
7.2.3.5.2 The dust used in the test described in 7.2.3.5.1 shall consist of about 1 kg of dry quartz with the particle size distribution shall be as follows:
a) passing through 150µm aperture, 104µm wire diameter 99 % to 100 %;
b) passing through 105µm aperture, 64µm wire diameter: 76 % to 86 %; and
c) passing through 75µm aperture, 52µm wire diameter: 60 % to 70 %.
7.2.4 Straps
7.2.4.1 Strap strength test
7.2.4.1-1 Carry out each test on two new samples of strap, conditioned as specified in 6.2.4.
7.2.4.1 .2 Grip each strap between the clamps of a tensile-strength testing machine. The clamps shall be so designed as to avoid breakage of the strap at or near them. The speed of traverse shall be about 1 00 mm/min. The free length of the specimen between the clamps of the machine at the start of the test shall be 200 mm ± 40 mm.
7.2.4.1.3 Increase the tension until the strap breaks and note the breaking load.
7.2.4.1 .4 If the strap slips or breaks at or within 1 mm of either of the clamps, deem the test to be invalid and carry out a new test out on another specimen.
7.2.4.2 Conditioning
7.2.4.2.1 Room conditioning
Keep the strap for 24 h ± 1 h in an atmosphere that has a temperature of 23ͦ C ± 5 ͦ C and a relative humidity of (50 ±10) %. If the test is not carried out immediately after conditioning, place the specimen in a hermetically closed receptacle until the test begins. Determine the breaking load within 5 min after removal of the strap from the conditioning atmosphere or from the receptacle.
7.2.4.2.2 Light conditioning
7.2.4.2.2.1 Use the apparatus described in SABS ISO 105-B02, Textiles^- Tests for colour fastness -Part B02: Colour fastness to artificial light « Xenon arc fading lamp test and a test strap of length at least 1,3 m.
Expose a central portion of the strap, of length at least 200 mm, to light for the time necessary to produce fading of standard blue dye No. 7 to a contrast equal to grade No, 4 on the grey scale according to SABS ISO 105-AQ2, Textiles - Tests for colour fastness - Part A02: Grey scale for assessing change in colour
7.2.4.2.2.2 After exposure, keep the strap for a minimum of 24 h in an atmosphere that has a temperature of 23 ͦ C ± 5 ̊ C and a relative humidity of (50 ± 10) %. Determine the breaking load within 6 min after removal of the strap from the conditioning apparatus.
7.2.4.2.3 Cold conditioning
7.2.4.2.3.1 Keep the strap for a minimum of 24 h in an atmosphere at a temperature of 23 ̊ C ± 5 °C and a relative humidity of (50 ± 10) %,
7.2.4.2.3.2 Then keep the strap for 90 min ± 5 min on a plane surface in a low-temperature chamber In which the air temperature is -30 *C ± 5 'C, Fold the strap and load the fold with a mass piece of mass 2 kg ±0,2 kg that has previously been cooled to -30 'C ± 5 'C. When the strap has been kept under load for 30. min ± 5 min in the same low-temperature chamber, remove the mass piece and measure the breaking load within 5 min after removal of the strap from the low-temperature chamber.
7.2.4.2.4 Heat conditioning
7.2.4.2.4.1 Keep the strap for 1 80 min ± 1 min in a heating-cabinet atmosphere that has a temperature of 60 ̊'C ± 5 ̊ C and a relative humidity of (65 ± 5) %.
7.2.4.2.4.2 Determine the breaking load within 5 min after removal of the strap from the heating cabinet.
7.2.4.2.5 Exposure to water
7.2.4.2.5.1 Keep the strap fully immersed for 1 80 min ±10 min in distilled water, at a temperature of 20 ̊ 0 ± 5 ̊ C, to which a trace of wetting agent has been added. Any wetting agent suitable for the fibre under test may be used.
7.2-4.2.5-2 The breaking load shall be determined within 10 min after removal of the strap from the water.
7.2.4.2-6 Abrasion conditioning
7.2.4.2.6.1 Keep the components or devices to be subjected to the abrasion test in an atmosphere at a temperature of 23 X ± 5 ̊ C and a relative humidity of (50 ± 10) % for a minimum of 24 h before testing. Keep the room temperature during testing between 15 °C and 30 °C.
7.2.4.2-6.2 Table 2 sets out the general conditions for each type of abrasion conditioning.
7.2.4.2.6.3 Where there is insufficient strap to test over 300 mm of shift, the test may be applied over a shorter length, subject to a minimum of 100 mm.
7.2.4.2 J Particular test conditions
7.2.4.2.7.1 Type 1 (in cases where the strap slides through the quick adjusting device)
Apply the 10 N load vertically and permanently on one of the straps. Attach the other strap, horizontally, to a device that is capable of giving the strap a back-and-forth motion. So place the adjusting device that the horizontal strap remains under tension (see figure N.I of annex N).
7.2.5.2.7.2 Type 2 (in cases where the strap changes direction in passing through a rigid part)
During this test, the angles of both straps shall be as shown In figure N.2 of annex N. The 5 N load shall be permanently applied. In cases where the strap changes direction more than once in passing through a rigid part, the load of 5 N may be so increased as to achieve the prescribed 300 mm of strap movement through that rigid part.
7.2.5 Lock-off devices
7.2.5.1 Class A devices.
Set up the child restraint with the largest manikin for which the child restraint is intended, as shown in figure 5. Use the webbing as specified in annex B of this specification. Fully apply the lock-off device and make a mark on the belt where the belt enters the lock-off device. Attach the force gauges to the belt via a D ring, and apply a force equal to twice (± 5 %) the mass of the heaviest dummy of group I
for at least 1 s. Apply the force for a further nine times. Make a further mark on the belt where it enters the lock-off device and measure the distance between the two marks. During this test, the retractor shall be unlocked.
7.2.5.2 Class B devices
Firmly secure the child restraint and pass webbing, as specified in annex 8 of this specification, through the lock-off device and frame following the routing described in the manufacturer's instructions. Pass the belt through the testing equipment, as shown in figure 6, and attach it to a mass of 5,25 kg ± 0.05 kg. Ensure that there is 650 mm ± 40 mm of free webbing between the mass and the point where the webbing leaves the frame. Fully apply the lock-off device and make a mark on the belt where it enters the lock-off device. Raise and then release the mass so that It falls freely over a distance of 25 mm ± 1 mm. Repeat this 1 00 times ± 2 times at a frequency of 60 cycles 1 2 cycles per minute, to simulate the jerking action of a child restraint in a car. Make a further mark on the belt where it enters the lock-off device and measure the distance between the two marks. The lock-off device shall cover the full width of the webbing in the installed condition with a 15 kg dummy installed. Conduct the test using the same webbing angles as those formed in normal use and with the free end of the lap belt portion fixed. For the duration of the test the child restraint system is firmly attached to the test bench used in the overturning or dynamic test. The loading strap can be attached to the simulated buckle.
7.2,7 Conditioning procedure for adjusters mounted direct on a child restraint
Install the largest dummy for which the restraint is intended, as if for the dynamic test, including the standard slack as specified in 7.1 .3.6. Mark a reference line on the webbing where the free end of the webbing enters the adjusting device.
Remove the dummy and place the restraint in the conditioning rig shown in figure P. 1 of annex P.
Cycle the webbing for a total distance of not less than 150 mm through the adjusting device. This movement shall be such that at least 100 mm of webbing on the side of the reference line towards the free end of the webbing and the remainder of the moving distance (approx. 50 mm) on the integral harness side of the reference line moves through the adjusting device.
If the length of webbing from the reference line to the free end of the webbing is insufficient for the movement described above, the 1 50 mm of movement through the adjusting device shall be from the fully extended harness position. The frequency of cycling shall be 10 ± 1 cycles/minute, with a velocity on S of 1 50 ± 1 mm/sec.
7.3.1 High-speed films and videos
7.3.1 Determine the behaviour of the test manikin and its displacement by means of a high-speed camera.
7.3.2 Firmly mount a calibration screen on the trolley or in the vehicle structure in such a way that the displacement of the manikin can be determined. - Reference of the measure
- Regulations 6.1.4.1 to 6.1.4.3
Regulation 7 - Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 9401.20 - Seats of a kind used for motor vehicles Yes Child restraint - Description
- Child restraint
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 7007: Safety glass, consisting of toughened (tempered) or laminated glass.
The measure came into effect on 16 July 2014
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 16 July 2014
- Publication where the measure is specified
- Government Gazette No. 37631, 16 May 2014
- Regulation where the measure is specified
- The Compulsory Specification For Safety Glass And Other Safety Glazing Materials - VC 9003
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- 1.1 This compulsory specification covers the requirements for safety glass and other safety glazing materials used in buildings in accordance with Part N of the National Building Regulations, and in any other application in which breakage due to human contact may result in serious injuries. Products within the scope of this compulsory specification include but are not limited to: Stock sheets of laminated safety glazing materials, i.e. sheets of safety glazing produced by a manufacturer equipped to laminate safety glass that will be cut to size by an installer;
• Toughened safety glass, i.e. panels of flat safety glazing manufactured from flat glass, cut to size, and then tempered;
• Polymeric glazing, i.e. sheets of transparent plastics that can be cut to size;
• Any glazed products that may be subjected to human impact;
• Any other material claimed to be safety glazing.
1.2 Safety glazing used in automotive applications is excluded. - Description of the measure
- A.1.3 Evidence of conformity including test reports issued not more than 12 months before the date of submission to the NRCS by a conformity assessment body recognized in terms of the NRCS's Conformity Assessment Policy, to prove compliance with all the relevant requirements of this compulsory specification;
- Reference of the measure
- Annex A.1.3
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 7007 Safety glass, consisting of toughened (tempered) or laminated glass. No - Description
- Safety glazing material: Glazing material that is so manufactured, constructed, treated and combined with other materials and components that, if broken by human contact, the likelihood of cutting or piercing injuries that might result from such contact are minimized.
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 7321.12: -- For liquid fuel and 7321.82: -- For liquid fuel
The measure came into effect on 08 October 2013
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 08 October 2013
- Publication where the measure is specified
- Government Notice No. R. 552 (Government Gazette No. 36725) Of 8 August 2013
- Regulation where the measure is specified
- Amendment of the Compulsory Specification For Non-Pressure Paraffin Stoves And Heaters -VC 9089
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This compulsory specification covers the requirements for the construction, operation and safe use of non-pressure paraffin stoves and heaters primarily intended for domestic use with a gauge pressure close to the burner < 2 kPa. gauge and with a maximum heat rating of 4,5 kW for a single burner.
- Description of the measure
- A1.2.2 Test report/s issued by an appropriately recognised conformity assessment body in a format acceptable to the NRCS and in accordance with the NRCS's Conformity Assessment Policy.
- Reference of the measure
- Annexure A1.2.2
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 7321.12 -- For liquid fuel Yes non-pressure paraffin stove 7321.82 -- For liquid fuel Yes non-pressure paraffin heater - Description
- A category of a non-pressure paraffin stove or heater that is similar in essential characteristics such as design,
materials, manufacturing processes or methods of assembly.
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 8701.90.90: -- Other
The measure came into effect on 20 April 2004
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 20 April 2004
- Publication where the measure is specified
- Government Notice R. 209 (Government Gazette 26014) Of 20 February 2004
- Regulation where the measure is specified
- Compulsory Specification For Agricultural Tractors
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- 1.1 This specification covers the requirements for agricultural tractors that have a maximum design speed not exceeding 40 km/h, and that have not previously been registered in South Africa.
1.2 This specification does not apply to:
a) experimental or prototype agricultural tractors that have been constructed or imported for the purpose of testing, assessment or development; and
b) industrial tractors, earthmoving tractors and purpose-built forestry tractors,
1.3 The relevant requirements of the specification that take effect on any specified date do not apply to
agricultural tractors manufactured or imported before the operative date given in schedule 1 of this specification.
1.4 Where a South African national standard, including an international standard or an ECE regulation adopted by Standards South Africa as a national standard, is incorporated by reference into this specification, only the technical requirements/specifications for the commodity, and the tests to verify compliance, apply. - Description of the measure
- 7 Proof of compliance
Homologation shall comprise the confirmation by the Regulatory Authority that the manufacturer or importer has provided the following specific evidence in respect of the commodity covered by this specification:
a) a summary of evidence that shows that all the relevant tests have been conducted with successful results under appropriate controls in respect of the model or variant of the commodity;
b) sufficient data to enable a relevant model or variant, and its components to be identified and related
to (a) above;
c) relevant samples for the conducting of whatever tests and inspections considered appropriate by the Regulatory Authority, to verify any or all of the evidence provided;
d) details of the qualify management system applied by the manufacturer or importer; and
e) when relevant, documentation to advise subsequent manufacturers or importers of incomplete commodities of their responsibilities.
The Regulatory Authority may issue such confirmation, on application, in respect of new models or variants, provided that such confirmation is not used for the purposes of advertising or to imply that all units of the commodity necessarily or consequently comply with all the requirements of the specification. - Reference of the measure
- Regulation 7.1
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 8701.90.90 -- Other Yes Agricultural tractors that have a maximum design speed not exceeding 40 km/h - Description
- Agricultural tractors that have a maximum design speed not exceeding 40 km/h, and that have not previously been registered in South Africa.
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 0307.81: -- Live, fresh or chilled
The measure came into effect on 15 August 2012
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 15 August 2012
- Publication where the measure is specified
- Government Gazette No.35436, 15 June 2012
- Regulation where the measure is specified
- The Introduction Of The Compulsory Specification For Live Aquacultured Abalone (VC 9001)
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- 1.1 This Compulsory Specification applies to the harvesting, preparation, packing, conveyance and quality of live aquacultured abalone.
1.2 Wild harvested live abalone is excluded from the scope of this Compulsory Specification. - Description of the measure
- 3.8 The testing of live aquacultured abalone against the requirements of this Compulsory Specification shall be done by test facilities that are accredited to use the test methods as referenced in the SAMSM&CP. In the case where there are no test facilities available that are in compliance with the foregoing, the NRCS will determine which facilities can be used in terms of its conformity assessment policy. This includes testing undertaken by farms and/or packers to demonstrate conformance with this Compulsory Specification.
4.5 All official sampling of live aquacultured abalone and water shall take place according to the requirements of the SAMSM&CP. - Reference of the measure
- Regulation 3.8 and 4.5
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 0307.81 -- Live, fresh or chilled Yes Live aquacultured abalone - Description
- Live aquacultured abalone
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa for 8535.2: - Automatic circuit breakers : and 8536.20: - Automatic circuit breakers
The measure came into effect on 16 December 1987
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 16 December 1987
- Publication where the measure is specified
- Government Notice 2286 (Government Gazette 10987) Of 16 October 1987
- Regulation where the measure is specified
- Compulsory Specification For Earth Leakage Protection Units
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This specification covers earth leakage protection units (circuit-breakers) for use in domestic and industrial or similar electrical installations and connected to a source of alternating current supply operating at a frequency of 50 Hz, the neutral conductor of which is connected to the general mass of earth on the supply side, and subject to the following limitations:
(a) Maximum rated voltage.................................................. . . . . . 500v.
(b) Maximum rated current ....................... ................................ 100 A.
(c) Maximum rated earth leakage tripping current.. ........................30mA
(d) Ambient temperature ..................... ...................................... - 5 ̊C to +10 ̊C.
The earth leakage protection units covered by this specification are intended to reduce fire and electrocution
hazards caused by current leaking from line and neutral conductors to earth.
This specification does not cover earth leakage protection units intended for service under extreme operating
conditions such as continuous high atmospheric humidity, atmospheric pollution, mechanical vibration. mechanical shock, and excessively high or low temperatures. - Description of the measure
- 6.2.1 Unless otherwise required, carry out the tests on each EL circuit-breaker as a whole and with the ambient
temperature maintained at 15-25 ͦͦ C.
6.2.2Test each single-pole and switched-neutral EL circuit-breaker, and each single-pole and neutral EL circuit-breaker, as a double-pole EL circuit-breaker.
6.2.3 With the exception of the air-core reactor in any phase (which shall be shunted by a resistance of a value such that the current through the resistor is 0,6 -0,7 % of the current through the air-core reactor), do not connect in parallel the reactance or resistance components of a test circuit load impedance.
Except where otherwise specified, use for the test an alternating current supply of practically sinusoidal wave form and that has a frequency of 50 +- 2 Hz. For any test where no tolerances are stated, the following general tolerances shall apply:
Voltage +- 5%; Current +- 5%; Frequency +- 2Hz; Time +- 5%
6.3 INSULATION RESISTANCE TEST: Measure at a d.c. voltage of 500 V the insulation resistance in the following positions:
(a) Between live terminals and any metal parts that would be exposed when the EL circuit-breaker is mounted in its normal position. Do this test (with all exposed metal parts electrically connected together and with all live terminals electrically connected together) with the EL circuit-breaker first in the "ON" position and then in the "Off" position.
(b) Between each incoming and each outgoing terminal, with the EL circuit-breaker in the "OFF" position
and, in the case of multiple circuit-breakers, with electrical interconnection between -
(1) all incoming terminals, and
(2) all outgoing terminals.
Check for compliance with 4.2.
6.6 OVERCURRENT RELEASE TEST.
6.6.1 Mounting and connections: Mount the EL circuit-breaker as for normal service (with all covers in position) and using conductors of length at least 1 m and of the size given in Table 2, appropriate to the current rating of EL circuit-breaker, connect it to the power supply.
6.6.2 Procedure:
(a) Using any convenient voltage, determine the current time charactersist of the EL circuit-breaker operatinf it at ambient temperature of 25+- 5 ̊C at all the following percentages of rated current:
135 1000
200 2000
400
Test each pole fitted with an overcurrent release separately except that for calibration at 135 %; of the rated current, all poles shall be tested simultaneously and shall be equally loaded. Test two poles. or one pole and the neutral, to ensure that the currents through the sensing device are balanced. Check for compliance with 4.5.1.3
Note: Where the characteristics of an EL circuit-breaker are such that the above points do not produce a
smooth curve. additional intermediate points may be taken as required.
(b) Finally, verify the current rating by operating the EL circuit-breaker at its rated through all poles
simultaneously for at least two hours.
6.7 OVERLOAD TEST FOR A N EL ClRCUIT-BREAKERS FITTED WITH AN OVERCURRENT RELEA SE.
6.7.1 Test circuit: With the EL circuit-breaker mounted as for normal service, connect it into a !circuit in which the current has been adjusted to 600+-5% of the relevant rated value and the applied voltage to 100 +-5% of the rated voltage of the EL circuit-breaker. Use an a.c. circuit having a lagging power factor of 0,45-0.5.
6.7.2 Procedure: Unless otherwise required, close and open the EL circuit-breaker 50 times consecutively at the rate of six operation cycles per minute and unsure that the contacts do not remain closed for more than two seconds in each cycle. In the case of multi-pole EL circuit-breakers, test simultaneously all poles intended to interrupt current, each pole carrying the current specified above. In the case of an EL circuit-breaker with no overcurrent release but fitted with a shunt release, carry out the test as described above but so energize the release (by applying a voltage equal to the rated voltage of the release) that the circuit-breaker opens 20-40 ms after closing. Check for compliance with 4.6.
6.8 OVERLOAD TEST FOR AN EL CIRCUIT-BREAKER NOT FITI'ED WITH AN OVERCURRENT RELEASE.
6.8.1 Test circuit: With the EL circuit-breaker mounted as for normal service, connect it into a circuit in which the current has been adjusted to 150% of the relevant rated value with a power factor of 0,95 and the applied voltage has been adjusted to 110+- 5 % of the rated voltage of the EL circuit-breaker.
Procedure: Close and open the EL circuit-breaker five times consecutively and ensure that the contacts do not remain closed for more than two seconds in each cycle. In the case of multiple EL circuit-breakers, test simultaneously all poles intended to interrupt current, each pole carrying the current specified above. Check for compliance with 4.6.
6.9 TEMPERATURE RISE TEST: Mount the EL circuit-breaker as for normal service (with all covers in position) in a place where there are no draughts and where the ambient air temperature is 25 +- 5 ̊C.
Using conductors each of length at least 1 m and of the size given in Table 2, appropriate to the current rating of the EL circuit-breaker, connect the EL circuit-breaker to the power supply and pass the rated current through the EL circuitbreaker for a period of two hours or until thermal equilibrium is attained. Then measure the relevant temperatures (see Table 4)by means of thermocouplesor any other acceptable method, at the points where the temperatures are judged to be highest. Check for compliance with 4.7.
6.10 MECHANICAL STRENGTH TEST.
6.10.1 Apparatus:
(a) A mechanical strength test apparatus as shown in Fig. 1. The metal cylinder A of mass 250 g and of
external diameter 25 mm fits loosely over the guide rod B. The guide rod is not rigidly fixed to the frame of the test apparatus and can be easily slid up and down. A hard fibre washer C ofdiameter25 mm and of thickness 12.5 mm is fixed to the bottom of the guide rod.
6.10.2 Specimen: Use one EL circuit-breaker from the sample.
6.10.3 Procedure:
(a) Place the hardwood block on the base of the test apparatus and f m l y hold or otherwise mount the
specimen on the block with its mounting surface down (i.e. toggle uppermost).
(b) Raise the guide rod and so move the block that, when the guide rod is lowered, the fibre washer rests on the surface of the specimen. Lower the guide rod, raise the metal cylinder to a height of 250 mm above the fibre washer, and allow the cylinder to drop onto the fibre washer.
(c) Repeat the drop twice, ensuring that the toggle is subjected to one drop when it is in the OFF position.
(d) So turn the specimen that it lies on its side on the block and proceed as described in (b) above, subjecting the specimen to three drops, two at random (but not on an edge of the specimen) and one approximately in the middle of the terminal guard.
(b)A rectangular hardwood block D that fits on the base of the test apparatus.
(e) Turn the specimen onto the other side and repeat the procedure given in (d) above.
(f)After the specimen has been subjected to nine drops, examine it for compliance with 4.8.
6.11 TEST FOR PERFORMANCE OF AN EL CIRCUIT-BREAKER UNDER EARTH FAULT CONDlTIONS:
Connect a 1 A fuse between the earthed conductor of the supply circuit and any metal parts that will be exposed when the EL circuit-breaker is mounted in its normal position. Connect all exposed metal parts together electrically. Connect the EL circuit-breaker to a power supply having an open circuit voltage of 100 2 5 % of the rated voltage.
Adjust the impedance of the load to provide a prospective earth fault current of 500 A with a power factor of 0,45-0,50. Cover that part of the surface of the EL circuit-breaker in which the toggle is situated with clean dry butter muslin that complies with requirement for Grade 1 butter muslin of SABS 446 'Absorbent gauze (fabric and swabs) and butter muslin'. Ensure that any cut or torn edges of the muslin are not exposed directly to the arc openings where flame may be emitted. Apply the earth fault current once by means of a separate switch in the supply circuit and once by means of the EL circuit-breaker switch, with the separate switch closed (i.e. close the EL circuit-breaker switch onto the fault.) In the case of a three-phase EL circuit-breaker test only one pole. Check for compliance with 4.9.
Immediately after the test, check the EL circuit-breaker for performance under earth leakage conditions at room temperature, the earth leakage current being applied gradually (see 6.14.3). Should the unit fail to comply with the requirement given in 4.12 (d), repeat the earth leakage test (as in 6.14.3) after 24 hours. - Reference of the measure
- Regulation 6.2.1 - 6.11
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 8535.2 - Automatic circuit breakers : No 8536.20 - Automatic circuit breakers No - Description
- Earth leakage protection units (circuit-breakers)
Countries/Regions affected by the measure.
- Description
- All countries
-
Applied by South Africa on the entire world for 3819.00: Hydraulic brake fluids and other prepared liquids for hydraulic transmission, not containing or containing less than 70 % by weight of petroleum oils or oils obtained from bituminous minerals.
The measure came into effect on 17 July 1975
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 17 July 1975
- Publication where the measure is specified
- Government Notice 128 (Government Gazette 4562) Of 17 January 1975
- Regulation where the measure is specified
- Compulsory Specification For Hydraulic Brake And Clutch Fluid
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- This specification covers fluid of the non-petroleum and the non-silicone types suitable for use in automotive
hydraulic brake and clutch systems. - Coded list of objectives
- X: For purposes n.e.s.
- Description of the measure
- 4. METHODS OF TEST.
4.1 EQUILIBRIUM REFLUX BOILING POINT AT
A PRESSURE OF 101. 325 kPa.
4.1.1 Apparatus (see Fig. 1 and 2).
(a) flask. — A 100 ml round-bottomed flask of heat resistant glass with a short neck having a 19/38 standard- taper, female ground-glass joint, and a 10 mm outside diameter side tube which enters the flask at such an angle as to permit the end of die thermometer bulb to be directly centred in the flask 6.5 mm from die bottom.
(b) Condenser. — A reflux, glass tube condenser of the water-cooled type having a jacket 200 mm in length. The bottom end of the condenser shall have a 19/38 standard- taper, drip-tip, and a male ground-glass joint.
(c) Boiling stones. — Silicone carbide grains of No. 8 grit.
(d) Thermometer.— A thermometer having a range of
— 5 to 300 °C and accurately calibrated for 7,62 cm immersion.
(c) Neat source. — A means of heating that will provide the healing and reflux rates specified in 4.1.2.
4.1.2 Procedure. — Place 60±1 ml of the fluid together with three silicone carbide grains in the flask, attach the condenser and insert the thermometer through the side tube so that its end is 6,5 mm from the centre bottom of the flask. Seal the thermometer to the side tube with a short length of rubber tubing. Mount the flask, and turn on the condenser water. The water supply temperature shall be not more than 28 °C and the temperature rise through the condenser shall be not more than 2 °C.
Apply heat at such a rate that the fluid is refluxing in 10+/-2 min at a rate of 1 to 5 drops per second. During the next 5±2 min period by adjusting the heat input, ensure that the rate of reflux is I to 2 drops of equilibrium reflux per second. Maintain the specified reflux rate for an additional 2 min and then read the temperature and record the barometric pressure. The temperature reading should be the average of four readings taken at 30 s intervals.
4.1.3 Recording. — Correct the temperature for thermometer error and for difference in atmospheric pressure between that recorded and 101,325 kPa by applying the appropriate correction given in Table 2. Record the corrected temperature rounded off to the nearest 0,5 °C as the equilibrium reflux boiling point at a pressure of
101,325 kPa.
4.2 FLUID STABILITY.
4.2.1 Stability at high temperature.
4.2.1.1 Apparatus.— As described in 4.1
4.2.1.2 Procedure. — Heat a new 60 ±1 ml sample of the original test fluid to a temperature of 185±2 °C by the procedure specified in 4.1. Maintain it at that temperature for 120±5 min and then determine the equilibrium reflux boiling point again by the procedure described in 4.1.
4.2.1.3 Boiling point change. — Take as the change in the boiling point the difference between the two corrected equilibrium reflux boiling points.
4.2.2 Chemical stability.
4.2.2.1 Apparatus. — As described in 4.1.
4.2.2.2 Procedure.— Mix 30±ml of the fluid with 30±1 ml of standard compatibility fluid. Determine the equilibrium reflux boiling point of this fluid mixture by the procedure specified in 4.1, applying heat to the flask at such a rate that the fluid is refluxing in 10±2 min at a rate of one to five drops per second. Record the maximum fluid temperature observed during the first minute after the fluid begins to reflux at the specified rate. Over the next 15±1 min, adjust and maintain the rate of reflux to one to two drops per second. Maintain a constant equilibrium reflux rate of one to two drops per second for an additional 2 min. Record the average of four
temperature readings taken at 30 s intervals as the final equilibrium reflux boiling point.
4.2.2.3 Change in temperature. — Take as the change in temperature of the refluxing mixture the difference between the two corrected equilibrium reflux boiling points.
4.3 WET EQUILIBRIUM REFLUX BOILING POINT AT A PRESSURE OF 101,325 kPa.
4.3.1 Apparatus. — (See Fig. 3).
(a) Glass jars. — Four straight-sided, cylindrical, screw top glass jars, each of capacity approximately 475 ml and with inside dimensions of height 100 mm and diameter 75 mm, with matching screw lids that will provide water and vapour-tight seals.
(b) Desiccators and covers.— Four bowl-form glass desiccators of inside diameter 250 mm, with matching tubulated covers fitted with No. 8 rubber stoppers.
(c) Desiccator plates. — Four perforated porcelain plates, without feet, glazed on one side, and of diameter 230 mm.
4.3.2 Reagents.
(a) Ammonium sulphate, reagent grade.
(b) Distilled water.
(c) Compatibility fluid corresponding in quality to the reference standard of the South African Bureau of Standards and with an equilibrium reflux boiling point of not less than 182 °C when tested in accordance with 4.1.
4.3.3 Procedure.
4.3.3.1 Lubricate the ground glass joints of the desiccators with silicone grease. Into each desiccator place 450± 25 g of the ammonium sulphate and add 125±10 ml of the distilled water. The surface of the resultant ammonium sulphate slurry shall lie within 45±7 mm of the top surface of each desiccator plate. Use a fresh charge of ammonium sulphate slurry for each test. Condition the charged desiccators with their covers on and stoppers in place at a temperature of 23±2 °C for at least 12 h before use.
4.3.3.2 Determine the water content of the compatibility fluid, using a method of determination that is sufficiently accurate to measure the mass of water added to samples of the compatibility fluid within approximately 15 per cent of the water added for additions up to 0,8 per cent (m/m), and within approximately 5 per cent of the water added for additions greater than 0,8 per cent (m/m).
4.3.3.3 Adjust the water content of the compatibility fluid to 0,50±0,5 per cent (m/m) and pour 100±1 ml of this fluid into cadi of two of the glass jars. Pour 100 ±1 ml of the test fluid into each of the remaining two glass jars. Immediately place the jars separately into the four charged and conditioned desiccators, replace the desiccator covers and maintain the temperature throughout the humidification procedure at 23±2 °C. At intervals, remove the rubber stopper in the top of each desiccator that contains the compatibility fluid. Use a long needled hypodermic syringe to withdraw a sample of not more than 2 ml from each jar and determine its water content in accordance with 4.3.3.2. Do not remove more than 10 ml in total of the compatibility fluid from each jar during the humidification procedure. When the average water content of the duplicates of the compatibility fluid roaches 3,50 ± 0.05 per cent (m/m), remove the two fluid test specimens from their desiccators and immediately cap each jar tightly. Determine the equilibrium reflux boiling points of the duplicate test specimens in accordance with 4.1. If the two results agree within 4 °C, record the average as the wet equilibrium reflux boiling point. If the two results differ by more than 4 °C, repeat the test and record the average of the four individual equilibrium reflux boiling points as the wet equilibrium reflux boiling point.
4.4 KINEMATIC VISCOSITY.
4.4.1 Apparatus. — A calibrated capillary type viscosity tube capable of measuring viscosity to within the limits of error given in Table 3.
4.4.2 Procedure. — Determine the kinematic viscosity of the fluid at —40 °C and at 100 °C taking particular care to avoid contamination of die fluid by condensation of atmospheric moisture during the determination at — 40 °C.
4.5 NEUTRALITY.
4.5.1 Apparatus. — A pH meter equipped with a calibrated glass electrode and a calomel reference electrode.
4.5.2 Procedure— Mix equal volumes of the fluid and an 80 per cent aqueous (distilled water) solution of ethanol having a pH value of 7,0±0,1. Distilled water with a pH value of 7,0d=0,l may be used instead of the alcohol solution if the fluid is miscible with an equal volume of water. Determine the pH value of the mixture at a temperature of 23± 5°C.
4.6 CORROSIVENESS.
4.6.1 Metal strips. — Prepare as follows from the metals listed in Table 1 three sets of corrosion test strips corresponding in composition and dimensions to the reference standard of the South African Bureau of Standards: With the exception of the tinned iron strips, clean the strips by abrading all surfaces with 320A waterproof
silicone carbide paper wetted with 95 per cent ethanol until all surface scratches, cuts, and pits have been removed from the strips. Use a new piece of silicone carbide paper for each different type of metal. With the exception of the tinned iron strips, polish the strips with No. 00 grade steel wool. Wash the strips, including the
tinned iron ones, with 95 per cent ethanol, dry them with a clean lint-free cloth, and place them for at least 1 h in a disiccator maintained at 23±5 °C. After polishing, handle the strips with clean forceps or tongs to avoid contamination. Determine the mass of each strip to the nearest 0,1 mg and assemble each set of strips on an uncoated steel cotter pin or bolt in the following order: tinned iron, steel, aluminium, cast iron, brass, and copper. Ensure that the strips are in electrical contact, and except for the cast iron strips bend them so that there is a gap
of 3,0±0,5 mm between adjacent strips for a distance of about 5 cm from the free ends of the strips. Measure the total exposed surface area of each strip. Immerse the strip assemblies in 95 per cent ethanol and then handle them only with clean forceps or tongs. Dry the assemblies with dried filtered compressed air and desiccate them at least 1 h before use.
4.6.2 Styrene-butadiene rubber cups.
(a) Take three styrene-butadiene wheel cylinder rubber cups corresponding in quality to the reference standard of the South African Bureau of Standards. Measure to the nearest 0,02 mm the base diameter of these cups along the centre line of the lettering on the cup and at right angles to this centre line. Take the measurements at least 0,4 mm above the bottom edge and parallel to the base of the cup. Do not use any cup for which the two measurements differ by more than 0,08 mm. Average the two readings of each cup.
(b) By means of a Micro-tester apparatus determine on the inner surface of the test cups the hardness of the cups in International Rubber Hardness Degrees.
4.6.3 Procedure. — Place one rubber cup, with the lip edge facing up, in each of three straight-sided round glass jars of capacity approximately 475 ml and inner dimensions of approximately 100 mm in height and 75 mm in diameter. Insert one metal strip assembly inside each cup with the pinned end in contact with the concavity of
the cup and the free end extending upward in the jar. Mix 760 ml of fluid with 40 ml of distilled water. Use this mixture to cover the metal strip assembly in each jar to a depth of approximately 10 mm above the tops of the strips. To close the jar use only tinned steel lids vented with a hole of 0,8±0,1 mm in diameter. Tighten the lids
and place the jars in an oven maintained at 100+2 °C for 120±2 hs. Allow the jars to cool at 23±5 °C for 60 to 90 min and then use forceps or tongs to take the strip assemblies out of the jars, removing loose adhering sediment by agitating each assembly in the fluid in its jar. Examine the strips and jars for adhearing crystalline deposits, disassemble the metal strips, remove adhearing fluid by flushing with water, and clean individual strips by wiping with a cloth wetted with 95 per cent ethanol. Examine the strips for evidence of etching and pitting. Place the strips in a desiccator maintained at 23±5 °C for at least an hour. Determine the mass of each strip to the nearest 0,1 mg. Determine the difference in mass of each metal strip and divide the difference by the total exposed surface area of the metal strip expressed in square centimetres. Average the results for the three strips of each type of metal. Immediately following (he cooling period use forceps or tongs to remove the rubber cups from the jars, removing loosely adhering sediment by agitation of the cup in the fluid in the jar. Rinse the cups in 95 per cent ethanol, air-dry them, and examine them for evidence of sloughing, blistering, and other forms of disintegration.
Determine the hardness and base diameters in accordance with 4.6.2 within 15 min after removal from the fluid. Examine the fluid-water mixture for jelling. Agitate the liquid in the jars to uniformly suspend any sediment, transfer a 100 ml portion of the liquid from each jar to a cone-shaped centrifuge tube [see 4.6.4.1 (b)] and
then determine percentages of sediment as described in 4.6.4. Measure as described in 4.5 the pH value of the fluid water mixture.
4.6.4 Determination of sediment.
4.6.4.1 Apparatus.
(a) Centrifuge. — A centrifuge capable of whirling two or more filled centrifuge tubes at a speed which can be controlled to give a relative centrifugal force of 600-700 at the tip of the tubes. Calculate the required speed of the rotating head by means of the following equation:
rpm = 423 squareroot(rcf/d)
where rcf = relative centrifugal force
d — diameter of swing, in centimetres, measured between lips of opposite tubes when in rotating position
(b) Centrifuge tubes. — Centrifuge tubes made of thoroughly annealed glass and having the dimensions given in Fig. 4 and distinct graduations.
4.6.4.2 Procedure.- Balance the two centrifuge tubes or pairs of tubes containing the liquid under lest with their respective trunion cups and place them on opposite sides of the centrifuge head. Then whirl them for 10 min at a rate sufficient to produce a relative centrifugal force of 600-700 at the tips of the whirling lubes. Repeat this
operation until the volume of sediment in each tube containing brake fluid remains constant for three consecutive readings.
4.6.4.3 Recording. Record the average volume of sediment at the bottom of the centrifuge lubes to an accuracy of at least 0,1 ml.
4.7 FLUIDITY AND APPEARANCE AT LOW TEMPERATURES.
4.7.1 At —40 °C- Place 100±2 ml of fluid in a 125 ml sample bottle of outside diameter 37d_0,05 mm and overall length 165±.2,5 mm. Stopper the bottle with a cork and place for 144±4 h in a bath maintained at — 40±2 °C.
Remove the bottle from the bath, quickly wipe the bottle with a clean lint-free cloth saturated with acetone or 95 per cent ethanol, determine the transparency of the fluid by placing the bottle against a hiding power chart corresponding in design to the reference standard of the South African Bureau of Standards and by observing the clarity of the contrast lines on the chart when viewed through every part of the fluid. Examine the fluid for stratification and sedimentation. Invert the bottle and determine the time in seconds required for the air bubble to travel to the lop of the fluid.
4.7.2 At — 50 °C. — Use the same procedure as described in 4.7.1 but keep the bottle for 6±0,2 h in a bath maintained at — 50±2 °C.
4.8 EVAPORATION.
4.8.1 Procedure.— Determine to the nearest 0,01 g the mass of each of four covered Petri dishes, approximately 100 mm in diameter and 15 mm high. Place 25±1 ml of
fluid in each of the four tared dishes, replace the covers, and redetermine the mass to the nearest 0,01 g. Determine the mass of fluid from the difference in masses of the filled and empty dishes. Remove the covers, invert them, place each dish inside its cover in a top-vented, gravity-convection oven at 100±2°C; maintain this
temperature for 46±2 hs. Remove the dishes from the oven, replace the covers, cool in a desiccator lo 23±5 °C for 1 h, and determine the mass of each dish. Return all the dishes to the oven for a further 24d;2 hs, cool and determine the mass as before.
If at the end of 72±4 hs the average loss by evaporation is less than 60 per cent (m/m), terminate the test. Otherwise, continue this procedure until equilibrium is reached as evidenced by mass loss of less than 0,25 g in 24 hs on each dish, or until seven days have elapsed, whichever occurs first.
4.8.2 Calculation. — Calculate the percentage of fluid evaporated from each dish, average the results, and record this figure as the loss by evaporation.
4.8.3 Quality of residue. — Examine the residue in the dishes at the end of 1 h at 23±5 °C for compliance with 3.9.2 by rubbing any sediment with a fingertip to establish the presence or absence of grittiness and abrasiveness.
4.8.4 Pour point of residue. — Combine the residue from all four dishes in a 125 ml sample bottle (as described in 4.7.1) and store vertically in a cold bath at — 5±1 °C for 60±10 min. Quickly remove the bottle and place it in a horizontal position. The residue must flow not less than 5 mm along the bottle within 5 s.
4.9 WATER TOLERANCE.
4.9.1 At — 40. °C— Mix 3,5±0,1 ml of distilled water with 100±1 ml of fluid and pour the mixture into a cone- shaped centrifuge tube (see Fig. 4) as described in 4.6.4.1
(b). Stopper the tube with a cork and place it for 24±2 h in a cold bath maintained at — 40±2 °C. Remove the centrifuge tube from the bath, and quickly wipe the tube with a clean lint-free cloth saturated with acetone or 95 per cent ethanol. Place the tube against a hiding power chart corresponding in design to the reference standard
of the South African Bureau of Standards, and observe the clarity of the contrast lines on the chart when viewed through the fluid as a whole. Examine the fluid for stratification and sedementation. Invert the tube and determine the time in seconds required for the air bubble to travel to the top of the fluid. The air bubble shall be considered to have reached the top of the fluid when the top of the bubble reaches the 2 ml graduation mark of the centrifuge tube.
4.9.2 At 60 °C— Place, for 24±2 h, the centrifuge tube and fluid used for the test given in 4.9.1 in an oven maintained at 60±2 °C. Remove the tube from the oven and immediately examine the contents for stratification. Then determine the percentage by. volume of sediment as described in 4.6.4.
4.10. COMPATIBILITY.
4.10.1 At -40 °C— Mix 50±0,5 ml of fluid with 50±0,5 ml of compatibility fluid corresponding in quality to the reference standard of the South African Bureau of Standards. Pour this mixture into a cone-shaped centrifuge tube (see Fig. 4) as described in 4.6.4.1 (b), and stopper with a cork. Place the centrifuge tube for 24±2 h in a bath maintained at — 40±2 °C. Remove the centrifuge tube from the bath and quickly wipe the tube with a clean lint-free cloth saturated with acetone or 95 per cent ethanol. Place the tube against a hiding power chart corresponding in design to the reference standard of the South African Bureau of Standards, and observe the clarity of the contrast lines on the chart when viewed through the fluid as a whole. Examine the fluid for stratification and sedimentation.
4.11 RESISTANCE TO OXIDATION.
4.11.1 Preparation of metal strips. — Prepare and clear as described in 4.6.1 three sets of aluminium and cast iron corrosion test strips corresponding in composition and dimensions with the reference standard of the South African Bureau of Standards. Determine the mass of each strip to the nearest 0,1 mg and assemble a strip of each
metal on an uncoated steel cotter pin, separating the strip at each end with a piece of tinfoil approximately 12 mm in area and between 0,02 and 0,06 mm in thickness. The tinfoil shall contain at least 99,9 per cent of tin and not more than 0,025 per cent of lead.
4.11.2 Preparation of test mixture. — Place 30± 1 ml of fluid in a test tube of diameter 22 mm and length 175 mm. Add 60±2 mg of reagent grade benzoyl peroxide and 1,5±0,05 ml of distilled water to the tube. Stopper the tube and shake the contents without allowing the solution to wet the stopper. Place the stoppered tube in an oven
at 70±2 °C for 120±10 min, shaking every 15 min, to dissolve the peroxide. Remove the tube from the oven and allow to cool to 23±5 "C. Use this test mixture not later than 24 h after removal from the oven.
4.11.3 Procedure. — Cut a styrene-butadiene wheel cylinder rubber cup into eight sections of approximately equal mass and place one section in the bottom of each of three test tubes 22 mm in diameter and 175 mm in length. Add 10 ml of the prepared test mixture to each test tube. Place a metal strip assembly in each lube with the free
ends of the strips resting on the rubber, the test mixture covering about one-half the length of the strips, and the end having the cotter pin projecting above the fluid. Stopper the tubes with corks and store them upright for 70±2 h at 23=h5 °C. Loosen the stoppers, place the tubes for 168±2 h in an oven maintained at 70 ±2 °C and then
remove and disassemble the metal strips.
4.11.4 Examination and calculation. — Examine the strips for gum deposits. Wipe the strips with a cloth soaked in 95 per cent ethanol and examine for pitting, etching or roughening of their surfaces. Place the strips for at least 1 h in a desiccator maintained at 23±5 °C, then determine the mass of each strip to the nearest 0,1 mg, and determine the loss caused by oxidation by dividing the difference in mass of each metal strip by the total exposed surface area of each metal strip expressed in square centimetres. Record, to the nearest 0,05 mg per cm 2 . The average of the results for the three strips of each type of metal "separately.
4. 12 EFFECT ON RUBBER.
4.12.1 Rubber cups. — Four styrene-butadiene wheel cylinder rubber cups corresponding in quality to the reference standards of the South African Bureau of Standards. Measure the base diameter and hardness of each cup as described in 4.6.2 fa) and 4.6.2 (b) respectively. Do not use any cup for which the two base diameter measurements differ by more than 0,08 mm.
4.12.2 Procedure.
(a) At 70 °C. — Place two rubber cups in a straight- sided cylindrical glass jar having a capacity of about 250 ml and inside dimensions of approximately 125 mm in height and 50 mm in diameter. Add 75 ml of fluid to the jar. Close the jar tightly with a tinned steel lid. Heal the jar for 120±2 h at 70±2 °C. Allow the jar to cool at 23±5 °C for 60-90 min. Remove the cups from the jars, wash the cups immediately with 95 per cent ethanol, and air-dry them. Examine the cups for disintegration as evidenced by stickiness, blistering, or sloughing. Within 15 min after removal from the fluid measure the base diameter and hardness of each cup as described in 4.6.2 (a) and 4.6.2 (b) respectively.
(b) At 120 °C— Place the other two rubber cups in a 250 ml glass jar with lid [see (a) above]. Add 75 ml of fluid to the jar and heat for 70±2 h at 120:fc2 °C. Allow the jar to cool at 23±5 °C for 60-90 min. Remove the cups from the jars, immediately wash the cups with 95 per cent ethanol, and air-dry them. Examine the cups for
disintegration as evidenced by stickiness, blistering, or sloughing. Within 15 min after removal from the fluid measure the base diameter and hardness of each cup as described in 4.6.2 (a) and 4.6.2 (b) respectively.
4.13 SIMULATED SERVICE PERFORMANCE.
4.13.1 Test apparatus and materials.— A stroking test apparatus (see Fig. 5) corresponding in design and component quality to the reference standard of the South African Bureau of Standards and consisting of the following:
(a) Master cylinder assembly. — A new hydraulic brake system cylinder that has an inside diameter of approximately 28 mm, a cast iron housing and is filled with an uncoated steel stand pipe. The piston shall be of half hard copper base alloy.
(b) Brake assemblies. — Four new straight bore hydraulic brake wheel cylinder assemblies in cast iron housings that have diameters of approximately 28 mm and pistons made from unanodized aluminium alloy. Each assembly shall have a forward brake shoe with lining, a reverse brake shoe with lining, a front brake drum assembly, and the necessary assembly components.
(c) Braking pressure actuating mechanism. — An actuating mechanism for applying a force, free from side- thrust, to the master cylinder push rod. The amount of force applied by the actuating mechanism shall be adjustable and capable of applying sufficient thrust to the master cylinder to create a pressure of at least 6 900 kPa in the simulated brake system. A hydraulic gauge or pressure recorder that has a range of at least 0-6 900 kPa shall be installed in the system and shall be provided with a shut-off valve and with a bleeding valve for removing air from the connecting tubing. The actuating mechanism shall permit adjustable "stroking rates of approximately 1000 strokes per hour. A mechanical or electrical counter shall be used to record the total number of strokes.
(d) Heated air cabinet.— An insulated cabinet or oven that has sufficient capacity to house the four mounted wheel cylinder assemblies, master cylinder, and necessary connections and having a thermostatically controlled heating system to maintain a temperature of 120:±5 °C. Heaters shall be shielded to prevent direct radiation to
wheel and master cylinders.
4.13.2 Preparation of test apparatus.
(a) Wheel cylinder assemblies. — Disassemble the cylinders and discard the rubber cups. Clean all metal parts with 95 per cent ethanol and dry with clean compressed air. Inspect the working surfaces of all metal parts for scoring, galling, pitting, and cylinder bore roughness, and replace all defective parts. Remove any stains on cylinder walls with coarse abrasive cloth and 95 per cent ethanol. If stains cannot be removed, replace the cylinder. Measure the internal diameter of each cylinder at four positions approximately 19 mm from each end of the cylinder bore by taking the measurements in line with the hydraulic inlet opening and at right angles to this
centre line. Replace the cylinder if any of these four readings exceed maximum or minimum limits of 28,66 or 28,60 mm respectively. Measure the outside diameter of each piston at two positions approximately 90 degrees apart. Replace any piston if either reading exceeds maximum or minimum limits of 28,35 or 28,52 nun respectively. Select the parts to insure that the clearance between each piston and mating cylinder is within 0,08-0,13 mm. Use the new styrene-butadiene rubber cups corresponding in quality to the reference standards of the South African Bureau uf Standards. Ensure that all cups are free from lint and dirt and do not use any cup which has defects such as cuts, moulding flaws, or blisters. Measure to the nearest 0,02 mm the lip and base diameter of all test cups along the centre line of I he lettering on the cup and at right angles to this centre line. Determine the
base diameter measurements at least 0,4 mm above the bottom edge and parallel to the base of the cup. Do not use any cup of which the lip or base diameters differ by more than 0.08 mm. Average the lip and base diameters of each cup. Determine the hardness of all cups by the procedure specified in 4.6.2 (b). Clean the rubber parts with 95 per cent ethanol and lint-free cloth. Dry with compressed air. With the exception of housings and rubber boots dip the rubber and metal parts of the wheel cylinders in the fluid to be tested and install them in the cylinders. Operate the cylinders manually to ensure that they operate easily. Install the cylinders in the simulated brake system.
(b) Master cylinder assembly. — Disassemble the master cylinder and discard all rubber components. Clean and measure all metal components as described in 4.13.2 (a). Use new styrene-butadiene rubber cups corresponding in quality to the reference standard of the South African Bureau of Standards which have been inspected and cleaned as described in 4.13.2 (a) and measured as described in 4.6.2 (a). Prior to determining the lip and base diameters of the secondary cup, clip the cup in the test fluid, assemble on the piston, and maintain die assembly at 23±5 °C for at least 12 h in a vertical position. Inspect the relief and supply ports of the master cylinder and replace the cylinder if these ports have burrs or wire edges. Measure the internal diameter of the cylinder at two positions: approximately midway between the relief and supply ports and approximately 19 mm beyond the relief port towards the bottom or discharge end of the bore, taking measurements at each position on the vertical and horizontal centre lines of the bore. Replace the cylinder if any reading exceeds maximum or minimum limits of 28,65 or 28,57 mm respectively. Measure each of the outside diameters of the master cylinder piston at two points approximately 90 degrees apart. Replace the piston if any of these four readings exceed maximum or minimum limits of 28,55 or 28,52 mm respectively.
Except for the housing and push rod-boot assembly, dip the rubber and metal parts of the master cylinder in the fluid to be tested and install them in the cylinder. Operate die master cylinder manually to ensure that it operates easily. Install the master cylinder in the simulated brake system.
(c) Assembly and adjustment of test apparatus. — With the wheel cylinder assemblies and master cylinder installed, adjust the brake shoe toe clearance to 1,0 ±0,1 mm. Fill the system with the test fluid and bleed all wheel cylinders and the pressure gauge to remove entrapped air from the system. Operate the actuator manually to apply a pressure of more than the required operating pressure, and inspect the system for leaks. Adjust the actuator to obtain a pressure of 6 900 .±.300 kPa. Adjust to a smooth pressure-stroke pattern and a stroking rate of 1 000±100 strokes per hour. Record the fluid level in the master cylinder stand pipe. 4.13.3 Test procedure.- Operate the system for 16 000 ±1000 cycles at 23 ±5 °C. Repair any leaks, re-adjust the brake shoe clearances, and add fluid to the master cylinder stand pipe to bring to the level originally recorded. Restart the actuating mechanism and within 6±2 h raise the temperature of the cabinet to 120±5 °C. Ensure that the wheel cylinders are functioning properly and record at intervals of 24 000 strokes during test the amount of fluid required to replenish any loss. Stop the test at the end of 85 000 total recorded strokes, i.e. including the number of
strokes during operation at 23± 5 °C and the number of strokes during the period required to bring the system to the operating temperature of 120± 5 °C. Allow the equipment to cool to room temperature. Examine the wheel cylinders for leakage. Operate the assembly for an additional 100 strokes, examine the wheel cylinders for leakage, and record the volume of any fluid lost. Discard the results of any test in which mechanical failure occurs which may affect the evaluation of the test fluid, and repeat the test.
4.13.4 Dismantling of apparatus and inspection of operating parts. — Within 16 h after completion of the test, remove the master and wheel cylinders from the system and retain the fluid in the cylinders by immediately capping or plugging the parts. Disassemble the cylinders and collect the fluid in a glass jar. Clean the rubber cups
in 95 per cent ethanol and dry them with compressed air. Inspect the cups for stickiness, scuffing, blistering, cracking, chipping, and change in shape. Within an hour after disassembly measure the lip and base diameters of each cylinder cup as described in 4.13.2 (a) and 4.13.2 (b), with the exception that the lip or base diameters may differ by more than 0,08 mm. Determine the hardness of each cup by the procedure specified in 4.6.2 (b).
4.13.5 Recording and calculation of test results.— Record any sludge, or jelling, present in the fluid after the test. Within an hour after draining the cylinders, agitate the fluid in the glass jar to suspend and uniformly disperse any sediment present and transfer a 100 ml aliquot of this sample to a cone-shaped centrifuge tube [see 4.6.4.1 (b), and determine the percentage by volume of sediment as described in 4.6.4. Allow the tube and fluid to stand for 24 h, recentrifuge and record any additional sediment recovered.
Inspect the cylinder parts and record any gumming or any pitting en pistons and cylinder walls. Rub any deposits adhering to the cylinder walls with a cloth wetted with 95 per cent ethanol to determine its abrasiveness and removability. Clean the cylinder parts in 95 per cent ethanol and dry with compressed air. Measure and record the diameters of the pistons and cylinders as described in 4.13.2 (a) and 4.13.2 (b). Calculate the lip diameter interference set from the following formula:
Lip diameter interference set, % =(D1-D2)/(D1-D3) *100
Where D1 + original lip diameter
D2 = final lip diameter
D3 = original cylinder bore diameter. - Reference of the measure
- Regulation 4
- Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 3819.00 Hydraulic brake fluids and other prepared liquids for hydraulic transmission, not containing or containing less than 70 % by weight of petroleum oils or oils obtained from bituminous minerals. Yes Hydraulic brake and clutch fluid - Description
- Hydraulic brake and clutch fluid
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
-
Applied by South Africa on the entire world for 8414.80: - Other
The measure came into effect on 05 April 2001
Non-Tariff Measure
- NTM classification
- B82: Testing requirement
- Date when the measure came into force
- 05 April 2001
- Publication where the measure is specified
- Government Notice R93 (Government Gazette 22014) Of 2 February 2001
- Regulation where the measure is specified
- Compulsory Specification For Microbiological Safety Cabinets (Classes I, II And III)
- Country/Region applying the measure
- South Africa
- The rationale of the measure
- 1.1 This specification covers requirements for the construction, fittings and pre-installation and post- installation performance of class I, class li and class III microbiological safety cabinets (also known as biological safety cabinets) intended to protect the operator and the environment from hazardous microbiological materials and (if so required by the customer or user or both), organic toxins and non-corrosive volatile organic agents.
NOTE - Microbiological safety cabinets are not intended to provide protection against corrosive chemical or radioactive materials.
1 .2 The specification does not cover the actual design of a safety cabinet and in no way restricts new design, provided that a microbiological safety cabinet of a new design complies with the requirements for materials, reliability, performance and safety given in this specification.
NOTE - Microbiological safety cabinets of class I, class II and class III should not be confused with laminar flow
clean workstations that usually discharge horizontally and vertically towards the operator and that do not provide
protection for an operator, but can even increase exposure to airborne hazards. - Coded list of objectives
- X: For purposes n.e.s.
- Description of the measure
- 5.2 Installation
After installation, the following tests shall be performed to ensure that the cabinet complies with the relevant performance and safety requirements.
a) Gastightness of the outer shell (class III cabinets)
When tested in accordance with 6.1, a class III cabinet shall comply with the requirements of 4.3.1.3.
b) HEPA-filter and HEPA-filter installation Integrity
When tested in accordance with 6.4, class I, class II and class III cabinets shall comply with the requirements of 3.5.1 .3.1 .
c) Integrity of the viewing window seal
When tested in accordance with 6.5, a class II cabinet shall comply with the requirements of 3.4.3.4.
d) Flow and distribution of air, average velocity and uniformity and rate of airflow
Class I cabinets: When determined in accordance with 6.6.3.1, the flow and distribution of air through the work-access aperture shall comply with the requirements of 4.1 .3.2.
When a smoke test is carried out in accordance with 6.5.3.3. the airflow shall comply with the requirements of 4.1 .3.1 .
Class 11 cabinets: When determined in accordance with 6.6.3.2.1, the velocity and uniformity of airflow in the work space shall comply with the requirements of 4.2.4.2.1 .
When determined in accordance with 6.6.3.2.2, the inward airflow velocity through the work-access aperture shall comply with the requirements of 4.2.4.3.3.
Class III cabinets: When determined in accordance with 6.6.3.3, the airflow shall comply with the requirements of 4.3.4.1 and 4.3.4.2.
6 Methods of test
6.1 Determination of gastlghtness of outer shell (class ill cabinets)
6.1.1 Principle
The cabinet is sealed and positively pressurized with hydrofluorocarbon gas. All surfaces and joints are scanned with the detector probe for leakage of the gas.
6.1.2 Apparatus
6.1.2.1 Gas detector, adjusted and calibrated to detect, at a reference leak source, the loss of hydro- fluorocarbon gas at a maximum rate of 16,5 g per annum.
6.1.2.2 Manometer, with scale divisions not exceeding 10 Pa and that is capable of
registering pressures m the range 200 Pa to 300 Pa.
6.1.2.3 Cylinder of hydrofluorocarbon gas (1.1.1.2 tetra-fluoro-ethane), commercially available as a refrigerant, R1 34(a), with a regulator valve, nozzle and connecting hose.
6.1.3 Procedure
6.1 .3.1 Prepare the cabinet for testing as a closed system by sealing all openings such as the exhaust opening, removable panel and other penetrations by any convenient means. Remove all external covers that are not essential for the operation of the cabinet.
6.1.3.2 Attach the manometer to the relevant test area of the cabinet to indicate interior pressure.
6.1.3.3 Suitably connect the gas cylinder to the test area and release the gas to positively pressurize the cabinet interior to a pressure of 250 Pa ± 10 Pa.
6.1.3.4 Prepare, calibrate and operate the gas detector in accordance with the manufacturer's instructions.
6.1.3.5 Move the probe of the instrument over the seams, joints, utility penetrations, gaskets and other locations of possible leakage, keeping the probe 7 mm to 12 mm from any surface and moving it at a rate of approximately 0,013 m/s.
6.1.4 Evaluation
Deem the cabinet to be gaslight if at no location a gas leak in excess of 1 6,5 g per annum is detected.
6.2 Determination of illuminance
6.2.1 Principle
Measurements of illuminance are taken at evenly spaced locations at a specified work level.
6.2.2 Apparatus
Illuminance meter (calibrated, cosine and vision-corrected), of such range that the illuminance measured is at least one-fifth of the full-scale value.
6.2.3 Procedure
6.2.3.1 Operate the lamps in the cabinet for at least 2 h.
6 2.3.2 Take eight illuminance measurements at eight evenly spaced locations at a height not exceeding 25 mm from the surface of the work floor but not within 1 50 mm of the perimeter of the work space. Record the results obtained at each location.
6.3 Determination of vibration
6.3.1 Principle
Measurements of the vibration velocity are made with a simple vibration meter at the geometric centre of the work surface, both with and without the cabinet in operation, to permit comparison of the vibration levels under these two conditions. Determination of the net vibration, i.e. vibration attributable to the cabinet alone, would require vibration frequency analysis.
6.3.2 Apparatus
Vibration meter, capable of measuring steady-state vibration velocities in the range
0,05 mm/s-1 ,0 mm/s (r.m.s.) in the frequency range 10 Hz ± 1 Hz to 250 Hz ± 25 Hz.
6.3.3 Procedure
6.3.3.1 Attach the vibration meter to the geometric centre of the work surface.
6.3.3.2 Ensure that the airflow is as specified.
6.3.3.3 With the cabinet in normal operation, measure the gross vibration velocity in the vertical, horizontal front to rear, and horizontal side-to-side axes.
6.3.3.4 Turn off the mechanical system and with the sensing element positioned and attached as in 6.3.3.1 , measure the ambient vibration velocity.
NOTE - The vibration frequency components of the ambient vibration are usually quite different from those of the mechanical cabinet system and hence the derivation of the net r.m.s. velocity (that attributable to the cabinet equipment) from measurements of gross and ambient vibration is not necessarily a simple mathematical subtraction.
6.3.4 Report
The following information shall be reported:
a) maximum value of the gross r.m.s. vibration velocity; and
b) maximum value of the ambient r.m.s. vibration velocity.
6.4 Determination of HEPA-filter and HEPA-filter installation integrity and
integrity of gaskets and construction joints in the vicinity of the HEPA-filter
installation
6.4.1 Principle
A polydisperse aerosol at ambient temperature is fed into the upstream side of the HEPA-filter installation at a specified flow rate and the downstream surface sides of the filters, seals, gaskets and construction joints in the vicinity of the filter installation is scanned with a probe nozzle to determine the percentage of penetration.
6.4.2 Apparatus
6.4.2.1 Vane anemometer or thermo-anemometer, as appropriate, accurate to within 2 %.
6.4.2.2 DOP generator
6.4.2.2.1 A cold DOP aerosol generator fitted with suitable nozzles and using filtered, compressed air at a pressure of 140 kPa ± 14 kPa. with the free airflow adjusted to not less than 30 e/min per nozzle producing an aerosol of particles with a median diameter of less than 0,8 pm.
6.4.2.2.2 A flexible aerosol delivery hose or tubing of nominal internal diameter 50 mm.
NOTES
1. A hot DOP aerosol generator that uses compressed nitrogen gas may be used to
determine the HEPA-filter installation integrity, provided that the aerosol particles produced should comply with the provisions in 6.4.2.2.1.
2 Liquids other than DOP may be used to generate an aerosol of particles, provided that the aerosol generator and the photometer have been suitably adjusted and calibrated for the alternative liquid. The aerosol produced should have a similar particle size distribution to that given in 6.4.2.2. 1.
6.4.2.3 Aerosol photometer
A light-scattering mass concentration indicator fitted with a probe nozzle. The probe nozzle or tip used for filter integrity testing is of internal diameter (d) not exceeding 30 mm. Any transition from initial inlet diameter to final inlet diameter is gradual. A maximum excluded angle of 20° is recommended. Photometers that have a threshold sensitivity of at least 10^3µg/l for DOP particles of diameter 0,3 pm, and that are capable of measuring concentrations in the range of 80 µg /l to 120 µg /l are suitable. The test photometer has a sample flow rate of 30 C/min ± 3 «/min. The probe inlet is of sufficient size to maintain the probe inlet rate at or slightly higher than a test flow rate of 27,5 «/min through the filter.
6.4,3 Procedure
NOTE - The test operator should avoid inhalation and exposure to heavy concentrations of the test aerosol. It is recommended that a suitable mask or respirator be worn for the duration of the test and thereafter, if necessary.
6.4.3.1 By using the method given in 6.6, determine the airflow and ensure that the flow through the air filter bank is within the operating limits of the cabinet design flow (see 4.1.3.2, 4.2.4.2. 4.3.4.1, or 4.3.4.2, as applicable). Ensure that the cabinet is operating normally while this procedure is being carried out.
6.4.3.2 Regulate the generator pressure and the gas flow rate in accordance with the manufacturer's instructions or as specified in 6.4.2.2.1 , as appropriate.
Introduce the aerosol via a sparge arrangement, if necessary, so that it is evenly distributed across the air entry.
6.4.3.3 For photometers that have
a) a linear readout, establish the upstream concentration by introducing the least amount of DOP aerosol required to produce a 100 % reading, thus allowing the instrument to be stray light adjusted to zero on the lowest scale range when the sample air stream is filtered free of aerosol, and
b) a logarithmic readout, adjust the upstream concentration (as determined from the instrument calibration curve) by introducing the least amount of DOP aerosol required to produce a concentration of 1 X 1 0^4 above that concentration required to give a reading of one scale division. Avoid prolonged exposure of filters to DOP.
6.4.3.4 With any removable filter guard removed, scan the entire filter media face in slightly overlapping strokes at a distance between the probe and the filter media face of approximately 25 mm and at a traverse rate not exceeding 50 mm and ensure that each filter pleat is scanned parallel to the direction of the pleat. Also scan the entire periphery of the filter at the bond between the filter media and the frame at the seal between the filter frame and the cabinet, and any construction joints
downstream of the filter installation. Record any local areas or points where a reading exceeding 0,03 % is obtained.
6.5 Method for the detection of leaks Into the work space of, and demonstration
of the integrity of a class II cabinet and its air barrier
6.5.1 Principle
6.5.1.1 While air-generated DOP aerosol is directed at joints in the vicinity of the work space or at the work-access aperture (air barrier), measurements are made using an aerosol photometer. Any meter readings in excess of 0,03 % penetration are an indication of seal or joint leakage or induction of contaminants into the clean work zone.
6.5.1.2 Smoke is released on the work space side of the work-access aperture.
Escape of smoke to the ambient air indicates an ineffective air barrier.
6.5.2 Apparatus
6.5.2.1 DOP generator, as in 6.4.2.2.
6.5.2.2 Aerosol photometer, as in 6.4.2.3.
6.5.2.3 Air current tube (smoke generating tube).
6.5.2.4 Barrier test fitting. (For barrier integrity testing, the aerosol delivery hose is fitted with a smooth parallel-bore fitting of internal diameter 50 mm ± 1 mm at the point of discharge. The fitting is of overall length 250 mm ± 5 mm, incorporates flow straighteners at its inlet and has a square cut end as illustrated in figure 2.)
6.5.3 Procedure
NOTE - The test operator should avoid inhalation and exposure to heavy concentration
of the tests aerosol. It is recommended that a suitable mask or respirator be worn for the duration of the test and thereafter if necessary.
6.5.3.1 Joints and seals
6.5.3.1 .1 Using the aerosol photometer, measure the ambient aerosol level of the work room/environment and of the work space of the cabinet. If the reading of the aerosol photometer is less than 10^ 3 above the filter face reading, discharge sufficient aerosol at the exterior of the joint or seal from a distance of approximately 150 mm to ensure the challenge is maintained at 0,1 % concentration or more, with the photometer setting as that used to establish the 100 % baseline during the integrity testing of the HEPA-filter.
6.5.3.1 .2 Use the photometer to scan all construction joints bordering the work space Hold the probe nozzle inside the cabinet, not more than 25 mm away from the joint and move it along the joint at not more than 5 cm/s (see figure 3).
6.5.3.1.3 Start scanning approximately 3 s after the aerosol cloud has been directed at the joint.
6.5.3.2 Air barrier
6.5.3.2.1 Discharge sufficient cold aerosol at a distance of approximately 1 50 mm in front of each test position and scan the lower edge of the viewing glass with the probe inlet held inside the work space at a distance of 25 mm away from the glass and at 100 mm centres. Start scanning at a point 25 mm from each of the work surface boundaries (see figure 4).
6.5.3.2.2 Direct the probe inlet towards the work-access aperture and position its centre approximately 1 mm above the lower edge of the viewing window.
6.5.3.2.3 Operate the aerosol photometer at each test position for at least 1 5 s. Where an intermittent penetration reading greater than 0,03 % above the ambient concentration is obtained, continue for at least a further 30 s.
6.5.3.2.4 Scan the front edge of the work floor (not the front edge of the cabinet) at a distance of 25 mm and at 1 00 mm centres. Commence scanning at a point 25 mm from each of the work surface boundaries (see figure 5). Direct the probe inlet towards the work-access aperture and ensure that its centre is positioned approximately 25 mm above the work floor.
6.5.3.2.5 Operate the photometer at each test position for at least 15 s and record any photometer reading, and its location, in excess of 0,03 % aerosol penetration, relative to the 100 % measured upstream. Where an intermittent penetration reading greater than 0,03 % above the ambient concentration is obtained, continue for at least a further 30 s.
6.5.3.3 Smoke test
6.5.3.3.1 In addition, carry out a simple smoke test to determine the direction of airflow near tile work- access aperture. Generate aerosol or any other smoke on the ambient side of the aperture so that the smoke cloud is within 150 mm of the entire area of the opening and note the direction of the airflow.
6.5.3.3.2 Using the air current tube (smoke generating tube) (see 6.5.2.3), release an even stream of smoke at a distance of 70 mm ± 5 mm from the inside plane of the work-access aperture at a series of positions with the tip of the air current tube approximately 25 mm below the bottom edge of the viewing window. Test positions at approximately 50 mm from both inner side walls of the work space of the cabinet, and at intervals of approximately 100 mm between these points.
6.6 Determination of flow and distribution of air, average velocity and uniformity and rate of airflow
6.6.1 Principle
Airflow velocity readings are taken at selected locations, using an anemometer to determine the average airflow velocity, the uniformity of airflow and the airflow rate.
6.6.2 Apparatus
6.6.2.1 Vane anemometer, free-standing, where applicable, of appropriate vane diameter and accurate to within ± 2 %.
6.6.2.2 Thermo-anemometer, where applicable, and accurate to within ± 2 %.
6.6.2.3 Barometer.
6.6.2.4 Thermometer.
6.6.3 Procedure
6.6.3.1 Class I cabinets
Ensure that the cabinet is operating normally. With the vane anemometer in the plane of the work- access aperture, take and record velocity readings of the air flowing into the aperture for at least 1 min and at least five locations evenly distributed across the plane of the work-access aperture.
6.6.3.2 Class II cabinets
6.6.3.2.1 Down flow
6.6.3.2.1.1 Ensure that the cabinet is operating normally. Take and record velocity readings in the horizontal plane 100 mm above the top edge of the work-access aperture using a free-standing vane anemometer.
6.6.3.2.1.2 Take velocity readings at 200 mm to 225 mm intervals in both directions, starting at a location 75 mm to 1 00 mm from the inner edge of the work surface. Record each reading and its location.
6.6.3.2.1 .3 Record the pressure drop across the filter system as indicated by the manometer or gauge fitted to the cabinet.
6.6.3.2.2 inflow
6.6.3.2.2.1 Ensure that the cabinet is operating normally.
6.6.3.2.2.2 Using a suitable vane anemometer or thermo-anemometer (as applicable), take readings at multiple points on a plane bounded by the perimeter of the exhaust aperture or duct. Record each reading and calculate the mean airflow velocity.
6.6.3.2.2.3 Multiply the area of the exhaust aperture or duct by the mean velocity obtained to yield the volumetric discharge rate of effluent air.
6 6 3 2.2.4 Obtain the average inward airflow velocity at the work-access aperture by dividing the volumetric effluent air volume by the cross-sectional area of the work-access aperture.
6.6.3.3 Class III cabinets
6.6.3.3.1 Inflow through open glove ports
6.6.3.3.1.1 Ensure that the cabinet is operating normally.
6.6.3.3.1 .2 Remove the gloves. Using the vane anemometer placed at the centre of each open glove port, take and record the airflow velocity for at least 1 min.
6.6.3.3.2 Inflow through the inlet filter
6.6.3.3.2.1 Ensure that the cabinet is operating normally with the gloves attached.
6.6.3.3.2.2 Using the thermo-anemometer, take multiple measurements within the exhaust duct along two axes perpendicular to each other, and record the mean airflow velocity.
6.6.3.3.2.3 Multiply the area of the exhaust duct by the average velocity obtained to yield the volumetric discharge rate of effluent air which is equal to the inflow through the inlet filter.
6.6.4 Report
6.6.4.1 Class I
Report
a) each velocity reading and its location;
b) the average of the velocity readings taken;
c) maximum and minimum velocity readings; and
d) percentage variations from the average of the maximum and minimum readings.
6.6.4.2 Class II
6.6.4.2.1 Downflow
Report
a) the pressure drop across the filter system;
b) each velocity reading and its location;
c) the average of the velocity readings taken;
d) maximum and minimum velocity readings; and
e) percentage variations from the average of the maximum and minimum readings.
6.6.4.2.2 Inflow
Report
a) the pressure drop across the exhaust filter system;
b) exhaust airflow velocity (m/s);
c) dimensions of the exhaust duct and work-access aperture; and
d) mean inward airflow velocity (m/s).
6.6.4.3 Class III
Report
a) inflow velocity, right glove port (m/s);
b) inflow velocity, left glove port (m/s);
c) exhaust airi'low velocity (m/s);
d) dimensions of exhaust duct; and
e) inlet airflow rate (m^/min).
6.7 Determination of noise level
6.7.1 Principle
Noise levels are measured at selected locations near the cabinet under normal operating conditions
and the background ambient conditions are also recorded.
6.7.2 Apparatus
6.7.2.1 Sound level meter
Use an integrating sound level meter configuration, that complies at least with the accuracy requirements specified for a type 1 instrument in SABS IEC 60651:1979, Sound level meters, and SABS IEC 60804: 1 985, Integrating-averaging sound level meters, as published by Government Notice No. 399 of 1 April 1999. Use a windscreen of a type specified by the sound level meter's manufacturer as being suitable for the particular microphone and that does not detectably influence the accuracy of the meter under the ambient conditions of the test.
NOTE - In principle, no time weighting other than l-time weighting is allowed during Integration: S-time weighting in particular should be disabled when L is measured since it could introduce errors over short integration intervals.
6.7.2.2 Calibration source
As the calibration source, use a sound calibrator that complies with the requirements prescribed for a type 1 calibrator in SABS IEC 60942:1997, Electroacoustics - Sound calibrators, as published by Government Notice No. 399 of 1 April 1999.
6.7.3 Procedure
6.7.3.1 Ensure that the cabinet is operating normally. For class 1 and class 11 cabinets, measure and record the noise level with the sound level meter situated 0,3 m from, and 0,3 m above, the top edge of the work-access aperture, at the vertical centre line of the cabinet and 1 m from any other part of the cabinet including the duct work and from the discharge point of the extraction system, if fitted. For class III cabinets, take measurement with the sound level meter situated 0,6 m above the work surface of the cabinet and 0,3 m from the cabinet front, at the vertical centre line of the cabinet and 1 m from any duct work and from the discharge point of the extraction system, if fitted.
6.7.3.2 Ensure that the airflow of the cabinet is as specified. Take all measurements with the sound level meter set to use the A-weighted network and fast response. Using the acoustic calibration source, check the acoustic sensitivity of the sound level meter before and immediately after the measurements are made and discard the results If the two checks do not coincide to within 1,0 dB.
6.7.4 Report
Include the following details in the report:
a) all operating noise level measurements and their location;
b) the identified maximum noise level and its location; and
c) the ambient noise level measurements at locations where indicated.
6.8 Determination of the protection factor for class I and class II cabinets
6.8.1 Principle
6.8.1 .1 Tests which are performed to assure that aerosols will be contained within open-fronted micro- biological safety cabinets, are specified in terms of an operator protection factor. This factor expresses the leakage from the open front of a cabinet, of a given aerosol that was released within the world space.
6.8.1 .2 The transfer index defines the exposure experienced at a given point as a result of the release of a known amount of tracer substance (bacterial spores or potassium iodide particles), within the cabinet. This exposure is defined as n/(Ns), where N is the number of particles released and n is the number of particles recovered at a sampling rate of s, the sampling being continued to completion. The transfer ratio index with and without the cabinet defines the protection factor and it is necessary to
define the reference situations which represent the open bench conditions, that is the exposure that an operator is subjected to by working in a ventilated room without the use of a safety cabinet. The reference open-bench conditions are defined as a room with a ventilation rate V, of 10 m^3/min, with complete mixing. The transfer index of the reference room is equal to 1/V = 1/10 and the protection factor then becomes (Ns)/(10n) if the sampling rate s is expressed in cubic metres per minute, or
(Ns)/(10^4n), if s is expressed in litres per minute. The minimum value of the protection factor that can be determined depends on the sensitivity and selectivity of the test, i.e. the magnitude of the challenge, N, the sampling rate s, and the least number of particles recovered that can be readily distinguished from background contamination. Practical values for these are N at least 3 x 10^8, s at least 50 ?/min and n not exceeding 10, which leads to a minimum verifiable value of not less than 1,5 x10^5 for the protection factor.
6.8.1.3 An "artificial arm" in the form of a cylinder of diameter between 60 mm and 65 mm, is used to mimic the turbulence produced by the worker's arm at the front aperture.
6.8.1.4 Containment tests on safety cabinets can be performed
a) with a microbiological aerosol that consists of a fine spray of microorganisms produced by a nebulizer charged from an aqueous suspension, or
b) with an airborne challenge of potassium iodide particles produced by a spinning disk aerosol generator.
6.8.2 Microbiological method
NOTE - In a room where cross-contamination, external contamination or protection factor tests have recently resulted in considerable leakage of the bacterial challenge into the ambient air, it is particularly advisable to perform a background test for the presence of the test organism 24 h before performing protection factor tests. A count of more than five test organisms on one of the culture plates following a 1 min test should be regarded as unsatisfactory, and the protection factor tests should be postponed until the ambient air is no longer contaminated with the test organisms. It is advisable to perform the protection factor tests before the cross-contamination and
external contamination tests.
6.8.2.1 Apparatus and materials
6.8.2.1.1 Spore suspension
A suspension of spores of a non-pathogenic microorganism, for example Bacillus subtills var. globigii (SABS Type Culture Collection (SABSTCC) Sac 35) in sterile distilled water, standardized to contain approximately 10^8 spores to 10^9 spores per millilitre
6.8.2.1.2 Culture plates
Petri dishes of diameter 90 mm that contain 15 mHo20 mi of nutrient agar (see 6.8.2.1.5).
6.8.2.1 .3 Slit air samplers
Two slit air samplers, each being able to operate at between 25 i and 30 H of air per minute.
6.8.2.1.4 Nebulizer
A Collison six-jet nebulizer with an internal outlet of diameter 14 mm, operated from a pressure line at 70 kPa, that sprays approximately 0,2 ml/min and discharges not more than 10l/min of free air at a velocity of 0,8 m/s.
6.8.2.1.5 Nutrient agar
6.8.2.1.5.1 Ingredients
Agar………..15,0 g
Peptone……… 10, Og
Beef extract …….5,0 g
Sodium chloride…….. 5,0 g
Water ………1 000 ml
6.8.2.1.5.2 Procedure
Dissolve the ingredients in the water by heating. Adjust the pH value to 7,2. Sterilize in bulk by autoclaving at 1 21 °C for 1 5 min. Cool to 45 °C and aseptically dispense 1 5 m5 in sterile Petri dishes. Ensure that the surfaces of the Petri dishes are dry before use.
6.8.2.1.6 Ml agar
6.8.2.1.6.1 Ingredients
Nutrient broth 3,125 g
Manganese sulphate tetrahydrate 0,03 g
Dipotassium hydrogen phosphate 0,25 g
Agar (Oxoid no. 3) 12,0 g
Water sufficient to produce 1 000 mH
6.8.2.1.6.2 Procedure
Dissolve the ingredients in the water by heating. Dispense 30 mH volumes in medical flat bottles or 150 m2 volumes in Roux flasks. Sterilize by autoclaving at 121 °C for 1 5 min. Cool to 45 °C and place the medical flat bottles on a 1-in-4 sloped surface and the Roux flasks on a flat surface. Allow the agar to solidify.
6.8.2.1.7 Cylinder
A cylinder of length approximately 1 m and of diameter 60 mm to 65 mm, that has a smooth surface and is closed at both ends.
6.8.2.2 Preparation of Bacillus subtilis var, globigii spores
6.8.2.2.1 Prepare about 20 Ml agar slopes (see 6.8.2.1 .6) in medical flat bottles or in Roux flasks, as required, to yield the appropriate amount of spores.
6.8.2.2.2 Use a fresh culture of Bacillus subtilis var. globigii that had been subcultured for three days at 36 °C ± 1 °C.
6.8.2.2.3 Inoculate the agar slopes with the organism and incubate for one week at 36 °C ± 1 °C and then at room temperature until 80 % sporulation has been obtained (usually within about 10 d).
6.8.2.2.4 Make a spore stain of the culture after about eight days to determine the percentage of spores. If the percentage is less than 80 %, leave the cultures until 80 % sporulation has been obtained.
NOTE - Use a 5 % aqueous malachite green stain for determining the percentage of spores. Make a smear on a glass microscope slide and heat fix it. Place the slide over a small beaker of boiling water. (Rest the slide on the rim of the beaker.) Add malachite green fire to the slide. Leave on for five minutes, then wash off with water. Counter stain with safranine for approximately 30 s. Rinse off with water. Check under the microscope.
6.8.2.2.5 Once 80 % sporulation has occurred, very gently wash the culture off the slopes by means of a sterile glass rod and suspend the spores In 10 m? of sterile distilled water. Do not get pieces of agar in the suspension since this will allow the spores to germinate.
6.8.2.2.6 Centrifuge the suspension in sterile tubes with the tops covered with brown paper. Wash the spores three times with sterile distilled water, i.e. decant the supernatant and add fresh sterile water to remove all traces of the medium. Centrifuge each time for 20 min.
6.8.2.2.7 After washing, resuspend the spores in sterile water and heat shock at 60 °C for 30 min or at 70 °C for 20 min for three days in succession.
6.8.2.2.8 Prepare tenfold serial dilutions of spore suspensions and plate out 0,1 m? samples of each dilution on nutrient agar plates (see 6.8.2.1 .5) and incubate the plates at 36 °C ± 1 °C for 24 h to 48 h.
Count the number of colonies on those plates with between 30 colonies and 300 colonies. From this result, calculate the concentration of spores per millilitre of suspension.
6.8.2.2.9 Store the stock culture (2 x 10^ spores to 4 x 10^ spores/mfi) at 4 °C until needed.
6.8.3 Procedure for determining protection factor
6.8.3.1 General
For cabinets of width up to 1 m, carry out five replicate protection tests at the centre of the work-access aperture. For cabinets of width exceeding 1 m but not exceeding 1,9 m, carry out five replicate protection tests at the centre of the aperture, and five each at the centres of the right and left halves of the aperture respectively. In order to avoid confusion from background contamination, carry out the tests in a well-ventilated room, after estimating background contamination.
6.8.3.2 Precede this entire procedure by a control run with the nebulizer switched off.
6.8.3.3 Introduce the cylinder through the work-access aperture of the cabinet to disturb the airflow (to simulate an operator's arm). Centre the cylinder between the side walls of the safety cabinetwork space and, where appropriate, at the centre of the right and left halves of the work access aperture, and normal to the plane of the aperture, extending from the back of the work space to protrude at least 250 mm into the room from the plane of the aperture. Raise the lower surface of the cylinder to
between 65 mm and 75 mm from the cabinet floor.
6.8.3.4 Measurement of the concentration of spores in the spore suspension
6.8.3.4.1 Prepare tenfold serial dilutions of the stock spore suspension (see 6.8.2.2.9) and plate out 0,1 mm samples of each dilution on nutrient agar plates (see 6.8.2.1 .5).
6.8.3.4.2 Transfer a measured volume of the stock spore suspension (5 mUo 10 m?, as appropriate) into the nebulizer (see 6.8.2.1 .4) and weigh the nebulizer with its contents. After spraying, weigh the nebulizer again.
6.8.3.4.3 Prepare tenfold serial dilutions of the spore suspension that remains in the nebulizer and plate out 0,1 mH samples of each dilution on nutrient agar plates.
6.8.3.4.4 At least half of the original volume of the stock spore suspension shall remain in the nebulizer. Determine the volume as follows:
V+(M2-M1) (V/2)
where
M1 is the mass of the nebulizer plus contents before spraying, in grams;
M2 is the mass of the nebulizer plus contents after spraying, in grams; and
V is the volume of the initial spore suspension in the nebulizer, in millilitres;
and assuming the density of the spore suspension to be 1 ,0 g/m8.
6.8.3.4.5 Incubate both sets of inoculated plates (before and after spraying) at 36 °C ± 1 °C for a period of 24 h to 48 h and count the colonies on those plates that have between 30 colonies and 300 colonies on them.
6.8.3.4.6 From these counts, determine the concentration of spores per millilitre in the initial suspension n1 and in the final suspension n2.
6.8.3.5 Place the nebulizer inside the work space, and where appropriate, at the centre of the right and left halves of the work-access aperture, with its outlet or the appropriate extension thereof 100 mm behind the plane of the work-access aperture midway between the side walls of the work space, and directed towards the aperture, with the spray axis parallel to the work surface.
6.8.3.6 For class I cabinets (see figure 6(a)), ensure that the spray axis is below the cylinder and approximately midway between its lower surface and the work surface.
6.8.3.7 For class II cabinets (see figure 6(b)), ensure that the spray axis is level with the upper edge of the aperture.
Dimensions in millimeters
6.8.3.8 Position the slit air samplers outside the cabinet in front of the work-access aperture, with their inlets not more than 200 mm in front of the plane of the aperture. Ensure that the inlets are level with the top of the cylinder, one to the right and one to the left, and each not more than 150 mm from the axis of the cylinder.
6.8.3.9 Switch on the cabinet and allow it to run until normal operating conditions are reached. Adjust each sampler to a sampling rate of not less than 25 d/min and not more than 30 d/min. Place a nutrient agar plate (see 6.8.2.1 .5) in each slit air sampler. Start the samplers. After 30 s, start the nebulizer that contains the stock spore suspension. Run the nebulizer for a period of not less than 4 min to ensure
the dispersal of at least 3x10^ spores. Switch off the nebulizer and continue to run the samplers for a further 5 min.
6.8.3.10 Incubate the culture plates at 36 °C ± 1 °C for 24 h to 48 h.
6.8.3.11 After incubation, count the Bacillus subtilis var. globigii colonies. If there is no growth of Bacillus subtilis var. globigii colonies on the control plates, use the results for the calculation of the protection factor (see 6.8.3.14).
6.8.3.12 If there is growth of Bacillus subtilis var. globigii colonies on the control plates, clean the equipment and repeat the procedure.
6.8.3.13 Determine the challenge dose N, in spores per millilitre, using the equation
N=n2{M1-M2)-(n2-n1)V
where
M1 is the mass of the nebulizer plus contents before spraying, in grams;
M2 is the mass of the nebulizer plus contents after spraying, in grams;
n1 is the concentration of the initial suspension before spraying, in spores per millilitre;
n2 is the concentration of the suspension after spraying, in spores per millilitre; and
V is the volume of the initial spore suspension in the nebulizer, in miliilitres;
and assuming the density of the spore suspension to be 1,0 g/ml
6.8.3.15 Carry out five replicate protection tests. Ensure that no individual value of the protection factor is less than 1,0 x 10^5 (see 4.2.4.3.2).
6.8.4 Potassium iodide method
6.8.4.1 Materials and apparatus
6.8.4.1.1 Potassium iodide, 15g/S solution either in absolute ethanol or industrial methylated spirits with a water content of not more than 5 % (by volume).
6.8.4.1.2 Palladium chloride, 1,0 g/fi solution In 0,1 mol/l hydrochloric acid.
6.8.4.1.3 Aerosol generator assembly that comprises a 3,8 cm diameter spinning disc capable of rotating at 28 000 r/min ± 500 r/min, and a nozzle with a fine hole to deliver the potassium iodide solution (see 6.8.4.1.1) to the spinning disc, the gap between the end of the nozzle and the spinning disc being set to 0,1 mm; also a laboratory stand to hold the aerosol generator above the work surface when necessary.
6.8.4.1 .4 Air samplers, that work on a centripetal principle with a volume flow rate of air of 1 00 ? /min through the front orifice, and a cone that entrains some 3 % of this air, collecting approximately 1 00 % of any potassium iodide particles that enter the sampler; airflow through the samplers being provided by a centrifugal fan coupled to the air samplers by a fixed tube.
NOTE - The particles being heavier than air, follow a straight path through the cone and are deposited on a filter membrane located at the base of the cone, while air is deflected to the outside of the cone (see figure 7),
6.8.4.1 .5 Cylinder of length and diameter approximately 1 m and 60 mm to 65 mm. respectively, that has a smooth surface and is closed at both ends.
6.8.4.1.6 Petri dishes, of diameter 55 mm.
6.8.4.1.7 Filter membranes, of diameter 25 mm and with a pore size of 3 Mm.
6.8.4.2 Procedure
6 8 4 2 1 Set out two Petri dishes (see 6.8.4.1 .6) away from the cabinet being tested, one dish half filled with palladium chloride solution (see 6.8.4.1 .2) and the other one half filled with distilled water.
Replace the lids on each. Set out two filter papers for drying the filter membranes (see 6.8.4.1 .7).
6.8.4.2.2 Introduce the cylinder (see 6.8.4.1 .5) through the work-access aperture of the cabinet to mimic the turbulence produced by an operator's arm in front of the aperture. Centre the cylinder between the side walls of the cabinet work space and normal to the plane of the aperture, extending from the back of the work space to protrude at least 250 mm into the room from the plane of the aperture. Raise the lower surface of the cylinder to between 65 mm and 75 mm from the cabinet floor.
6.8.4.2.3 Place the aerosol generator (see 6.8.4.1 .3). inside the work space, with the leading edge of the disc 100 mm behind the plane of the front aperture.
6.8.4.2.4 For class I cabinets (see figure 6(a)), place the aerosol generator below the cylinder.
6.8.4.2.5 For class II cabinets (see figure 6(b)), ensure that the disc is level with the upper edge of the aperture.
6.8.4.2.6 Position two air samplers (see 6.8.4.1.4) outside the cabinet in front of the work-access aperture, with their inlets 1 50 mm to 160 mm in front of the plane of the aperture. Ensure that the inlets are level with the top of the cylinder, one to the right and one to the left, and each 150 mm from the axis of the cylinder.
6.8.4.2.7 Load each air sampler carefully with a filter membrane using a pair of fine-pointed forceps kept clean and dry solely for this purpose. Adjust the air pressure to 20 cm water gauge (this is consistent with an air sampling rate of 100 fi/min), using a U-tube water manometer one limb of which is attached to a pressure tapping on the rear of the sampler.
6.8.4.2.8 Measure 20 m? of potassium iodide (see 6.8.4.1 .1) into the aerosol generator reservoir with the fluid release valve closed.
6.8.4.2.9 Switch on the cabinet and allow it to run until normal operating conditions are reached. Apply suction to the air samplers and start the spinning disc. Wait 1 5 s and then open the release valve to allow the potassium iodide to feed on to the centre of the disc.
6.8.4.2.10 Turn off the air samplers after the generation of aerosol has stopped. Wait until the suction pump stops completely, and then remove the filter membrane from one sampler using a second pair of fine-pointed forceps designated for this purpose.
6.8.4.2.11 Gently float the filter membrane in the palladium chloride solution contained in the Petridish, with the surface that has been exposed to airflow facing upwards. Within 30 s to 45 s, the membrane will become saturated with palladium chloride and any potassium iodide particles will become visible as brown spots.
6.8.4.2.1 2 Remove the membrane with a third pair of fine-pointed forceps designated for this purpose, immerse the membrane in distilled water for 3 s to 4 s and then place it on a clean filter paper to dry.
Repeat this procedure with the filter membranes from the other air samplers. Replace the lids of the Petri dishes.
NOTE - The solution of potassium iodide used for the tests is flammable and corrosive to untreated steel; consequently the cabinet under test should be wiped clean with a wet cloth and scrupulous care should be taken with the spinning disc equipment.
6.8.4.2.13 Examine each filter with either a low-power binocular microscope or a 15 x magnifier and count the number of brown spots on the filter membrane.
6.8.4.2.14 Calculation of the protection factor
Calculate the number of potassium iodide particles released, N, using the following equation:
N = (3,1 X10^7x M)
where
3,1 X 10^7 is a constant derived from the droplet size, the sampling flow rate and the speed of rotation of the disc; and
M is the volume of potassium iodide solution dispersed by the aerosol generator, in
millilitres.
Then calculate a value for the protection factor, PF (separately for each filter membrane), using the
following equation:
PF = NV/10^4
where
V is the sampling flow rate in cubic metres per minute; and
n is the number of spots on the filter membranes.
NOTES
1 In this case, M is 20 ml (see 6.8.4.2.8) and Vis 100 l/min (see 6.8.4.2.7),
2 Using the above equations and the values of /Wand V given in note 1 , a protection factor of 1,0 x 10^5 would correspond to 62 spots on the filter membrane.
3 When calculating the protection factor, if there was only one spot on the filter membrane, the protection factor would be 2,6 X 1 0^6. If there were no spots on the filter membrane this would indicate that the protection factor was higher than this, and in the above example the protection factor would be recorded as PF > 2,6 x 10^6.
6.8.5 Background tests
Place two air samplers loaded with filter membranes in front of the safety cabinet, 1 50 mm to either side of the aperture centre line and 100 mm from the plane of the aperture. Turn on the sampler suction fan and run it for 10 min without any generation of potassium iodide droplets by the aerosol generator.
Remove the filter membranes and develop and examine them in accordance with 6.8.4.2.12 and 6.8.4.2.13.
NOTES
1 On completion of the background tests in laboratories where no previous tests have taken place within 24 h, the developed membranes should not show any brown spots.
2 In laboratories where protection factor tests have recently taken place (or where protection factor tests have resulted in a considerable leakage of aerosol challenge) it Is particularly advisable to perform background tests before further tests
on the cabinets. A count of more than five spots on one of the two filter membranes following a 1 min test should be regarded as unsatisfactory and further cabinet tests should be postponed until the background is no longer contaminated.
6.9 External contamination test
6.9.1 Principle
The integrity of the air barrier at the work-access aperture is indicated by the measurement of the inward penetration of bacterial spores which are sprayed into the opening with the cabinet operating normally.
6.9.2 Apparatus and materials
As in 6.8.2.1 (the slit air samplers are not required).
6.9.3 Procedure
6.9.3.1 Place the cylinder in the cabinet as described in 6.8.3.3.
6.9.3.2 Distribute at least 12 culture plates (see 6.8.2.1 .2) evenly over the work floor of the cabinet.
6.9.3.3 Load a measured volume of spore suspension into the nebulizer. Position the nebulizer outside the cabinet, with the nebulizer's delivery opening 100 mm in front of the centre of the top edge of the work-access aperture. Ensure that the spray axis is parallel to the work surface and directed into the cabinet.
6.9.3.4 Ensure that the cabinet is operating normally. Uncover the culture plates 1 min before spraying begins. Run the nebulizer for a period of at least 4 min to ensure the dispersal of a challenge dose of at least 3x10^6 spores. Switch off the nebulizer and leave the culture plates uncovered for a further 5 min.
6.9.3.5 In any test, the number of colonies of the test organism counted after incubation at 36 °C ± 1 °C for a period of 24 h to 48 h shall not exceed five. Carry out the test five times, using a new set of culture plates for each test.
6.9.3.6 Carry out a control test with the cabinet motor blower(s) switched off. Ensure that at least 300 colonies are recovered from these plates during the control test. If less than 300 colonies are recovered, repeat the test.
6.10 Cross-contamination test
6.10.1 Principle
Bacterial spores are sprayed across the work space and contamination of the opposite two-thirds of the cabinet is monitored.
6.10.2 Apparatus and materials
As in 6.8.2.1 (the slit air samplers are not required).
6.10.3 Procedure
6.10.3.1 Place the cylinder in the cabinet as described in 6.8.3.3.
6.10.3.2 Distribute at least 12 culture plates (see 6.8.2.1 .2) evenly over the right two-thirds of the work surface of the cabinet and at least 350 mm from the left side.
6.1 0.3.3 Load a measured volume of spore suspension into the nebulizer. Place the nebulizer with its spray axis 1 00 mm above the work surface and 50 mm from the left side of the work surface. Ensure that the spray axis is parallel to the work surface and directed towards the opposite wall.
6.10.3.4 Ensure that the cabinet is operating normally. Uncover the culture plates 1 min before spraying begins and cover them again 5 min after spraying stops. Run the nebulizer for a period of at least 4 min to ensure the dispersal of a challenge dose of at least 1 x 10^5 spores.
6.10.3.5 Count the number of colonies of the test organism after incubation at 36 °C ± 1 °C for a period of 48 h ± 2 h. Carry out the test three times.
6.10.3.6 Carry out the test three more times using reversed positions (i.e. placing the culture plates on the left and the nebulizer on the right).
6.10.3.7 Carry out a control test with the cabinet motor blower(s) switched off. Ensure that 300 colonies are recovered from these plates during the control test. If less than 300 colonies are recovered, repeat the test.
6.11 Test for resistance to corrosion
6.11.1 Test solution
6.11.1.1 Preparation of the sodium chloride solution
Dissolve a sufficient mass of sodium chloride in distilled or deionized water of conductivity not higher than 20 pS/cm at 25 °C ± 2 °C to produce a concentration of the sprayed solution collected of 50 gin ± 5 g/e. The specific gravity range for a 50 g/fi ± 5 g/H solution is 1,025 5 to 1,040 at 25 °C.
Ensure that the sodium chloride contains less than 0,001 % (by mass) of copper and less than 0,001 % (by mass) of nickel as determined by atomic absorption spectrophotometry or another analytical method of similar sensitivity. Ensure that the sodium chloride does not contain more than 0,1 % (by mass) of sodium iodide or more than 0,5 % (by mass) of total impurities calculated for dry salt.
If the pH of the prepared solution, measured at 25 °C ± 2 °C, is outside the range 6,0 to 7,0, investigate for the presence of undesirable impurities in the sodium chloride or in the water (or in both).
6.11.1.2 pH adjustment
So adjust the pH of the solution that the pH of the sprayed solution collected within the spray cabinet (see 6.11.2.1) is between 6,5 and 7,2. Check the pH using electrometric measurement at25 °C ±2 °C, or, in routine checks, with a short-range pH paper which can be read in increments of 0,3 pH units or less. Make any necessary correction by adding hydrochloric acid or sodium hydroxide solution of analytical grade.
Changes in pH can result from the loss of carbon dioxide from the solution when it is sprayed. Such changes can be avoided by reducing the carbon dioxide content of the solution by, for example, heating it to a temperature above 35 °C before it is placed in the apparatus, or by making the solution from freshly boiled water.
6.11.1.3 Filtration
If necessary, filter the solution before placing it in the reservoir of the apparatus (see 6. 11 .2.3), to remove any solid matter which might block the apertures of the spraying device.
6.11.2 Apparatus
Ensure that all components in contact with the spray or the test solution are made of, or lined with, materials resistant to corrosion brought about by the sprayed solution and which, in turn, do not influence the corrosiveness of the sprayed test solutions. The apparatus shall include the components given in 6.11.2.1 to 6.11.2.4.
6.11.2.1 Spray cabinet
The spray cabinet shall be of a capacity not less than 0,2 m^ and preferably not less than 0,4 m^, since, with smaller volumes, difficulties can be experienced in ensuring the even distribution of spray. For large-capacity cabinets, it is necessary to ensure that the conditions of homogeneity and distribution of the spray are met (see 6.1 1 .6). The upper parts of the cabinet shall be so designed that drops of
sprayed solution formed on its surface do not fall on the specimens being tested.
The size and shape of the cabinet shall be such that the collection rate of solution in the cabinet is within the limits specified in 6.1 1.6.3.
6.11.2.2 Heater and temperature control
An appropriate system that maintains the spray cabinet and its contents at the specified temperature (see 6.11 .6.1). The temperature shall be measured at least 100 mm away from the walls.
6.11.2.3 Spraying device
The device for spraying the sodium chloride solution comprises a supply of clean air at a controlled pressure and humidity, a reservoir to contain the solution to be sprayed, and one or more atomizers.
The compressed air supplied to the atomizers is passed through a filter to remove all traces of oil or solid matter and is at an absolute pressure of 70 kPa^1 to 170 kPa.
NOTE - Atomizing nozzles might have a "critical pressure" at which an abnormal increase in the corrosiveness of the salt
spray occurs. If the "critical pressure" of a nozzle has not been established with certainty, control fluctuations in the air pressure within ±0,7 kN/m^2, by installing a suitable pressure regulator valve to minimize the possibility of the nozzle being operated at its "critical pressure".
In order to prevent the evaporation of water from the sprayed droplets, the air is humidified before entering the atomizer, by passing through a saturation tower that contains hot water at a temperature several degrees Celsius higher than that of the cabinet. The appropriate temperature depends on the pressure used and on the type of atomizer nozzle and these shall be so adjusted that the rate of collection of spray in the cabinet and the concentration of the collected spray are kept within the specified limits (see 6.11.6). The level of the water is automatically maintained to ensure adequate humidification.
The atomizers are made of inert material, for example glass or plastics materials. Baffles may be used to prevent direct impact of spray on the test specimens and the use of adjustable baffles is helpful in obtaining uniform distribution of the spray within the cabinet. The level of the sodium chloride solution in the reservoir is automatically maintained to ensure uniform spray delivery throughout the test.
6.11.2.4 Collecting devices
At least two suitable collecting devices, which consist of funnels made of glass or other chemically inert material and with the stems inserted into graduated cylinders or other similar containers, shall be used.
Funnels of diameter 100 mm have a collecting area of approximately 80 cm^. The collecting devices are placed in the zone of the cabinet where the test specimens are placed, one close to an inlet of spray and one remote from an inlet. They are so placed that only spray, and not drops of liquid that fall from specimens or from parts of the cabinet, is collected.
6.11.2.5 Re-use
Ensure that if the equipment has been used for a spray test or for any other purpose and a solution different from the specified sodium chloride solution was used, it is thoroughly clean before use.
Operate the equipment for at least 24 h and measure the pH of the collected solution to ensure that it is correct throughout the entire spraying period, before any specimens are placed in the chamber.
6.11.3 Method of evaluation
To check the reproducibility of the test results for one piece of apparatus or for similar items of apparatus in different laboratories, it is necessary to verify the apparatus at regular intervals, as described in 6.11.3.1 to 6.11.3.4.
6.11.3.1 Reference specimens
To verify apparatus, use four reference specimens of thickness 1 mm ± 0.2 mm. width 50 mm and length 80 mm of CR4 grade steel that complies with SABS ISO 3574:1986, Cold-reduced carbon steel sheet of commercial and drawing qualities, as published by Government Notice No 399 of 1 April 1999 with a practically faultless surface^', and a matt finish (arithmetical mean deviation of the profile Ra = 1 .3 µm ± 0,4 µm). Cut these reference specimens from cold-rolled plate or strip.
Carefully clean the reference specimens immediately before testing. Besides the directions given in 6.11 .4.2 and 6. 11 .4.3, ensure that the cleaning eliminates all traces of dirt, oil and other foreign matter that could influence the test results.
Use one of the following methods:
a) clean the reference specimens by vapour degreasing using a chlorinated hydrocarbon. Use three successive treatments of 1 min each, with an interval of at least 1 min between successive treatments; or
b) thoroughly clean the reference specimens with an appropriate organic solvent (hydrocarbon, that has a boiling point between 60 °C and 120 °C) using a clean soft brush or an ultrasonic cleaning device. Carry out the cleaning in a vessel filled with solvent. After cleaning, rinse the reference specimens with fresh solvent, then dry them; or
c) other cleaning methods may be used after agreement between the interested parties, provided that the results will be comparable.
Determine the mass of the reference specimens to an accuracy within 1 mg. Protect one face of the reference specimens with a removable coating, for example, an adhesive plastic film.
6.11.3.2 Arrangement of the reference specimens
Position the four reference specimens in four different quadrants in the spray cabinet, with the unprotected faces upwards, and at an angle of 20° ± 5° from the vertical.
Use supports made of, or coated with, inert materials such as plastics. Ensure that the upper edges of the reference specimens are level with the top of the salt spray collector. The test duration is 96 h.
6.11.3.3 Determination of mass loss
At the end of the test, remove the protective coating. Remove the corrosion products by immersion in a cleaning solution of hydrochloric acid (^20 = 1,18 g/mO, of recognized analytical grade at 50 % (by volume), in water, inhibited by 3,5 g of hexamethylene tetramine per litre.
After stripping, thoroughly clean the reference specimens with water at ambient temperature, then with acetone, followed by drying.
Determine the mass of the reference specimens to an accuracy within the nearest 1 mg and calculate the mass loss of the exposed surfaces in grams per square metre.
6.11.3.4 Checking of apparatus operation
The operation of the test apparatus is deemed to be satisfactory if the mass loss of each reference specimen is 140 g/m^2 ± 40 g/m^2.
6. 11 .4 Test specimens
6.11.4.1 Two rectangular test specimens of nominal dimensions 100 mm x 15 mm of the metal used to manufacture the cabinet.
6.1 1 .4.2 Thoroughly clean the test specimens before testing (see 6.1 1 .3.1 ). The cleaning method used will depend on the nature of the material, its surface and the contaminants, but abrasives or solvents which might attack the surface of the specimens may not be used.
Ensure that specimens are not recontaminated after cleaning by careless handling.
Specimens intentionally coated with protective organic films should not be cleaned before the lest.
6.11.4.3 If the test specimens are to be cut from a larger coated article, ensure that the coating is not damaged in the area adjacent to the cut. Protect the cut edges by coating them with a material that will be stable under the conditions of the test, such as paint, wax or adhesive tape.
6.1 1 .5 Arrangement of the test specimens
6.1 1 .5.1 So place the test specimens in the spray cabinet that they are not in the direct line of travel of spray from the atomizer.
6.11.5.2 The angle at which the surface of a test specimen is exposed in the spray cabinet is very important. Support the specimen facing upwards in the spray cabinet at an angle as close as possible to 20° to the vertical but within the limits 15° to 30°. In the case of irregular surfaces, for example, entire components, adhere to these limits as closely as possible.
6.11.5.3 So arrange the test specimens that they do not come into contact with the cabinet and that surfaces to be tested are exposed to free circulation of spray. The specimens may be placed at different levels within the cabinet, as long as the solution does not drip from specimens or their supports at one level onto other specimens placed below. However, for a new examination or for tests with a total duration exceeding 96 h, location permutation of specimens is permitted.
6.1 1 .5.4 Ensure that the supports for the test specimens are made of inert non-metallic material such as glass, plastics or suitably coated wood. If it is necessary to suspend specimens, use synthetic fibre, cotton thread or other inert insulating material.
6.11.6 Operating conditions
6.1 1 .6.1 Maintain the temperature inside the spray cabinet at 35 °C ± 2 °C with the minimum possible fluctuation in temperature throughout the duration of the test.
6.1 1 .8.2 Start the test after it has been confirmed that the collection rate (see 6. 1 1 .6.3) and conditions (see 6.1 1 .6.1) are within the specified ranges, and the cabinet is filled with test specimens as planned.
6.11.6.3 Ensure that the solution collected in each of the collecting devices (6.1 1 .2.4) has a sodium chloride concentration of 50 g/l. ± 5 g/l and a pH value in the range 6,5 to 7,2.
Ensure that the average rate of collection of solution in each device, measured over a period of at least 24 h of continuous spraying is 1 mfi/h to 2 m?/h for a horizontal collection area of 80 cm^2.
6.11.6.4 Do not re-use test solution which has been sprayed.
6.11.6.5 During the test, prevent any increase or decrease of cabinet pressure.
6.11.7 Duration of test
6.1 1 .7.1 The duration of the test shall be at least 480 h.
6.11.7.2 Do not interrupt the spraying during the prescribed test period. Do not open the cabinet except for brief visual inspections of the test specimens in position and for replenishing the salt solution in the reservoir, if such replenishment cannot be carried out from outside the cabinet.
6.11.7.3 A periodic visual examination of specimens under test for a predetermined period is allowed, but do not disturb the surfaces under test and only open the cabinet for the minimum period necessary to observe and record any visible changes of the material under test.
6.11.8 Treatment of specimens after test
At the end of the test period, remove the test specimens from the cabinet and allow them to dry for 0,5 h to 1 h before rinsing, in order to reduce the risk of removing corrosion products. Before they are examined, carefully remove the residues of spray solution from the surfaces of the test specimens. A suitable method is to rinse or dip the test specimens gently in clean running water, at a temperature not exceeding 40 "'C and then to dry them immediately in a stream of air, at a pressure not exceeding
200 kPa and at a distance of approximately 300 mm.
6.11.9 Evaluation of results
The material shall be deemed to be corrosion resistant if corrosion of the base metal is not visible to the unaided eye.
6.11.10 Report
6.11.10.1 The report shall indicate the outcome of the test according to the criteria for the evaluation of results. Report the result obtained for each specimen tested and, when appropriate, the average result for a group of replicate test specimens. The report could be accompanied by photographic records of the tested specimens.
6.11.10.2 The following information shall be included in the report:
a) a reference to this compulsory specification;
b) the type and purity of sodium chloride and water used;
c) a description of the material or product tested;
d) the dimensions and shape of the test specimen and the nature and area of the surface tested;
e) the preparation of the test specimen, including any cleaning treatment applied and any protection given to edges or other special areas;
f) known characteristics of any coating with an indication of the surface area;
g) the number of test specimens subjected to the test and a reference to each material or product represented;
h) the method used to clean test specimens after the test, with, where appropriate, an indication of the loss in mass resulting from the cleaning operation;
i) the angle at which the tested surfaces were inclined;
j) the frequency and number of specimen location permutations, if any;
k) the duration of the test and the results of any intermediate inspections;
I) the test temperature;
m) the volume of collected solution;
n) the pH of the test solution and the collected solution;
o) the density of the collected solution;
p) any abnormality or incident that occurred during the entire test procedure; and
q) intervals of inspection.
6.12 Prefilter tests
Use the relevant test methods given in SABS 1424, Filters for use m air-conditioning and general ventilation, as published by Government Notice No. 1 851 of 1 December 1 995, to determine the initial arrestance.
Use a 600 mm x 600 mm sample of the prefilter in its unpleated state.
6.13 Chemical resistance
6.13.1 Reagents
Use the following reagents for the testing of chemical resistance:
hydrochloric acid, 40 g/l^3
sodium hydroxide, 40 g/l
quaternary ammonium compound, 10 g/ l
formaldehyde, 50 g/l.
sodium hypochlorite, 5 g/ l
iodophor, 20 g/ l
phenol, 50 g/ l
ethanol, 70 % (by volume)
6.13.2 Procedure
6.13.2.1 Apply approximately 0,5 m? of each reagent to the surface that has to be tested. Cover each reagent with a watch glass in the centre of the puddle with the concave side down and leave the reagent on the surface for 4 h, ensuring that the test surface is wet throughout the entire period.
6.13.2.2 After the 4 h period, scrub the surface with a stiff brush and hot water at a temperature of 70 °C.
6.13.2.3 Dry the surface and remove any surface stains by washing with alcohol before examination.
6.13.3 Inspection
Inspect the test surfaces for any visible effect other than a slight change of gloss or discoloration when compared with an untreated area.
6.14 Stability test
6.14.1 Principle
The stability test is performed to demonstrate the resistance of a microbiological safety cabinet to overturning under an applied force.
6.14.2 Apparatus
6.14.2.1 Compression force gauge, calibrated in kilograms, accurate to within ± 5 % full scale, or
6.14.2.2 Extension spring balance, calibrated in kilograms, accurate to within ± 5 % full scale.
6.14.3 Procedure
6.14.3.1 Block the cabinet at the front bottom edge to prevent lateral movement. Apply force, as appropriate, at the centre of the rear top edge (see figure 8) to subject the cabinet to a torque of 700 Nm.
6.14.3.2 Measure the cabinet lift at the rear bottom edge.
7 Blower performance test
7.1 Principle
The airflow velocity is increased to determine whether the motor blower has spare capacity.
7.2 Apparatus
7.2.1 Vane anemometer or thermo-anemometer (or both) as appropriate, accurate to within 2 %.
7.2.2 Timer.
7.3 Procedure
7.3.1 Class I
7.3.1.1 With clean HEPA-filters fitted, set the blower speed control so that when the inward airflow velocity is measured in accordance with 6.6.3.1 , the average airflow velocity shall be that specified in
3.6.2.1.
7.3.1.2 After 2 h, measure the average inward airflow velocity in accordance with 6.6.3.1 .
7.3.2 Class II
7.3.2.1 With clean HEPA-filters fitted, set the blower speed control so that when the downward airflow velocity in the work space is measured in accordance with 6.6.3.2.1, the average downward airflow velocity shall be that specified in 3.6.2.2.
7.3.2.2 After 2 h, measure the average downward airflow velocity in accordance with 6.6.3.2.1.
7.3.2.3 When a class II cabinet is fitted with a separate motor blower for the exhaust system, set the blower speed control so that when the inward airflow velocity through the work-access aperture is measured in accordance with 6.6.3.2.2, the average inward airflow velocity shall be that specified in 3.6.2.2.
7.3.2.4 After 2 h, measure the average inward airflow velocity in accordance with 6.6.3.2.2. - Reference of the measure
- Regulations 5.2
Regulations 6.6.1 to 7.3 - Measure also domestic
- Yes
Products affected by the measure.
Code Product Partial coverage Partial coverage indication Date in Date out 8414.80 - Other Yes Microbiological safety cabinets (classes I, II And II - Description
- Microbiological safety cabinets (classes I, II And III)
Countries/Regions affected by the measure.
Inclusion/Exclusion Country Date in Date out Inclusion Entire world - Description
- All countries
B82: Testing requirement
