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GB/T 21510-2008 (GB/T21510-2008)

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GB/T 21510-2008: PDF in English (GBT 21510-2008)

GB/T 21510-2008
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 11.080.99
G 70
Antimicrobial property detection methods for nano-
inorganic materials
ISSUED ON: MARCH 13, 2008
IMPLEMENTED ON: AUGUST 01, 2008
Issued by: General Administration of Quality Supervision, Inspection and
Quarantine;
Standardization Administration of the People's Republic of
China.
Table of Contents
Foreword ... 3 
1 Scope ... 4 
2 Normative references ... 4 
3 Terms and definitions ... 4 
4 Test methods ... 5 
5 Test data processing ... 5 
6 Calculation of detection results ... 5 
7 Detection report ... 6 
8 Precautions ... 6 
Annex A (normative) Test method for powder antimicrobial property - Oscillation
method ... 8 
Annex B (normative) Test method for material antimicrobial property -
Oscillation method ... 13 
Annex C (normative) Test method for material antimicrobial property - Filming
method ... 15 
Antimicrobial property detection methods for nano-
inorganic materials
1 Scope
This Standard specifies the terms and definitions, test methods, test data
processing, calculation of detection results, detection report and precautions for
antimicrobial property of nano-inorganic materials.
This Standard is applicable to nano antimicrobial powder and materials that use
nano antimicrobial powder as antimicrobial functional components (structural
units), such as fibers, fabrics, plastics, coatings and ceramics. The detection for
antimicrobial property of other materials can also refer to this Standard.
2 Normative references
The provisions in following documents become the provisions of this Standard
through reference in this Standard. For dated references, the subsequent
amendments (excluding corrigendum) or revisions do not apply to this Standard,
however, parties who reach an agreement based on this Standard are
encouraged to study if the latest versions of these documents are applicable.
For undated references, the latest edition of the referenced document applies.
GB/T 19619, Terminology for nanomaterials
GB/T 13221, Nanometer powder - Determination of particle size distribution
- Small angle X-ray
"Cosmetics Hygiene Standards" of the Ministry of Health of the People's
Republic of China (Edition 2002)
3 Terms and definitions
For the purposes of this document, the terms and definitions established in
GB/T 19619 as well as the followings apply.
3.1 antibacterial
the effect that can inhibit or hinder the growth and reproduction of bacteria or
fungi and their activity.
3.2 bactericidal
the effect that kills the growth and reproduction of bacteria or fungi.
3.3 antimicrobial
the effect that kills or hinders the growth and reproduction of bacteria or fungi
and their activity.
3.4 nano antimicrobial powder
an assembly of discrete nanoparticles that meets the requirements of GB/T
13221 and has antibacterial effects.
3.5 nano antimicrobial material
nano antibacterial powder and materials that use nano antibacterial powder as
antibacterial functional components (structural units).
4 Test methods
4.1 The test method for the antimicrobial property of nano powder shall be
carried out according to the method specified in Annex A.
4.2 The test method for the antimicrobial property of materials such as fibers,
fabrics, plastic powders and microporous filter materials shall be carried out
according to the method specified in Annex B.
4.3 The test method for the antimicrobial property of hard surface materials
such as plastics, ceramics, paint films, plates and metals are carried out in
accordance with the methods specified in Annex C.
5 Test data processing
Multiply the number of colonies on each plate by the dilution factor to obtain the
actual number of colonies recovered from the sample.
6 Calculation of detection results
6.1 Calculation of the number of colonies
Multiply the number of colonies on each plate by the dilution factor to obtain the
actual number of bacteria recovered from the sample.
Annex A
(normative)
Test method for powder antimicrobial property - Oscillation method
A.1 Scope of application
This test method is applicable to determination of the antimicrobial property of
nano powder.
A.2 Test equipment and materials
A.2.1 Test equipment
Type A2 secondary biological safety cabinet, constant temperature shaking
incubator (300r/min), constant temperature incubator, pressure steam sterilizer,
electric heating constant temperature dry oven [(0~250)°C], refrigerator,
microwave oven (output power ≥700W), balance (resolution of 0.001g).
A.2.2 Test equipment
Erlenmeyer flask (capacity of 500ml, 250ml, 150ml), petri dish (diameter of
90mm), test tube (18mm×180mm), graduated cylinder (100mL), straw (10mL,
5mL, 1mL), alcohol lamp, test tube rack and so on.
A.2.3 Medium and reagents
A.2.3.1 Ordinary nutrient broth medium
Peptone 10g
Beef extract 5g
Sodium chloride 5g
Distilled water 1000mL
Adjust the pH value to 7.2~7.4. Perform high-pressure steam sterilization at
121°C for 20min.
Usage: for the cultivation of staphylococcus aureus and Escherichia coli
enrichment.
A.2.3.2 Ordinary nutrient agar medium
Peptone 10g
Wash off the lawn. Transfer the washed bacteria liquid to another test tube.
After mixing with a shaker, use 0.03mol/L phosphate buffer to dilute to an
appropriate concentration (about 105cfu/mL). The suspension of bacterial
propagules shall be stored in a refrigerator at 4°C for later use and shall not
exceed 4h.
A.3.2.2 Preparation of control sample solution
Weigh 0.5g±0.05g of control sample powder into an Erlenmeyer flask. Add 95ml
of phosphate buffer containing 0.1% Tween-80. After mixing, add 5.0mL of pre-
prepared bacteria suspension.
A.3.2.3 Preparation of test group sample solution
Weigh 0.5g±0.05g of test sample powder into an Erlenmeyer flask. Add 95mL
of PBS containing 0.1% Tween-80. After mixing, add 5.0mL of pre-prepared
bacteria suspension.
A.3.2.4 Count of live bacteria in the control sample "0"
Before oscillation, properly dilute the control sample solution. Pipette 1.0mL and
inoculate it in a sterile dish. Inoculate 2 plates in parallel for each sample
solution. Pour the nutrient agar medium that has been melted at 45°C~55°C.
Turn the plate over after the agar medium has solidified. Place the above plate
in a 37°C±1°C constant temperature incubator for colony count.
A.3.2.5 Oscillation contact cultivation
Fix the Erlenmeyer flask containing the control sample and the test sample on
the shaking bed of the constant temperature shaking incubator. Under the
condition of action temperature 37°C±1°C, at the speed of 150r/min, oscillate
and contact the material that test sample needs to be diluted before use 1h~4h;
oscillate and contact the material that does not need to be diluted and is directly
used 4h~24h.
A.3.2.6 Viable bacteria count after oscillation for a certain period of time
After appropriate dilution of oscillated control sample solution and test solution,
respectively take 1.0 mL of the sample solution and inoculate it in a sterile dish.
Inoculate 2 plates in parallel for each sample solution. Pour the nutrient agar
medium that has been melted at 45°C~55°C. Turn the plate over after the agar
medium has solidified. Place the above plate in a 37°C±1°C constant
temperature incubator and count the live bacteria culture.
A.3.2.7 Negative control group
The negative control group draws the same batch of dilutions, culture medium
Annex B
(normative)
Test method for material antimicrobial property - Oscillation method
B.1 Scope of application
This test is applicable to the determination the antimicrobial property of
dissolvable and non-dissolvable fibers, fabrics, plastic powders, and
microporous filter materials.
B.2 Test equipment and materials
B.2.1 Test equipment, test device and standard strains for test
The requirements for test equipment, test device and standard strains for test
shall meet the provisions of A.2.1~A. 2.4.
B.2.2 Control sample
The control sample is a piece of pure cotton plain white cloth (32 yarns). The
sample itself has no antibacterial effect and has no influence on the
determination of test results. Degreasing treatment shall be carried out before
the test: Boil the pure cotton plain weave cloth in water with detergent for 30min;
Rinse 3 times with tap water; Then boil it with distilled water for 5min, rinse, dry
and iron; Before cutting, remove the warp and weft according to the size of the
prepared sample, and then cut it according to the draw marks.
B.3 Test procedures
B.3.1 Preparation of strain slope
The preparation of the strain slope shall comply with the provisions of A.3.1.
B.3.2 Test steps
B.3.2.1 Cut the antimicrobial fabric sample into 10mm×10mm. Antimicrobial
plastics, microporous filter materials, filaments, staple fibers, and yarns are
used as they are. Weigh 1.0g±0.05g of antimicrobial sample into an Erlenmeyer
flask. Add 95 mL of PBS containing 0.1% Tween-80. After mixing, add 5.0mL of
pre-prepared bacteria suspension.
B.3.2.2 Preparation of bacterial suspension, preparation of control sample
solution, count of viable bacteria of control sample "0" contact time, oscillation
contact culture, viable bacteria count after oscillation contact for a certain period
the bacterial suspension from overflowing.
C.3.2.2 Control sample
The control sample is injection-molded with sanitary high-density polyethylene
(HDPE). The standard size is 50mm×50mm (±2 mm). The thickness is not more
than 5mm. It is required that it has no antibacterial effect and has no influence
on the judgment of test results.
C.3.2.3 Preparation of test group samples
Make the test sample into a standard size of 50mm×50mm (±2mm). If the test
sample size is small, it shall not be less than 20mm×20mm.
C.3.2.4 Sample pretreatment
Take control samples and tested samples. Use 70% ethanol solution to wipe
the surfaces. After 5min, use sterile distilled water to rinse. Dry naturally. If the
sample is not suitable for disinfectant treatment, it can be directly rinsed with
sterile distilled water or disinfected by other methods according to the
characteristics of the sample. But it must not affect its antimicrobial property
and interfere with the test results.
C.3.2.5 Preparation of bacterial suspension
Take the nutrient agar medium slope from the third to the eighth generation of
strains for 18h-24h fresh culture. Use a 5.0mL pipette to suck 3.0mL~5.0mL of
0.03mol/L phosphate buffer into the inclined tube. Repeatedly suck and blow.
Wash off the lawn. Transfer the washed bacteria liquid to another test tube.
After mixing with an oscillator, use 0.03mol/L phosphate buffer to dilute to an
appropriate concentration (about 105cfu/mL). The suspension of bacterial
propagules shall be stored in a refrigerator at 4°C for later use and shall not
exceed 4h.
C.3.2.6 Inoculation bacteria solution
Put the control sample and the tested sample into a sterile dish. Pipette
0.2mL~0.5mL test bacteria solution and drip it on the surface of control sample
and tested sample respectively. Make 3 parallel samples for each sample. Use
a sterile tweezer to pick up the cover film and cover the sample surface
separately and flatten it. There must be no bubbles. Make the bacteria solution
evenly contact the sample. Cover the plate. Conduct contact cultivation at
37°C±1°C and relative humidity of 90% for 16h~24h. If the tested sample is a
photocatalyst antibacterial agent, a light source shall be installed in a constant
temperature incubator according to the sample test requirements.
C.3.2.7 Colony count
......
 
(Above excerpt was released on 2020-09-26, modified on 2021-06-07, translated/reviewed by: Wayne Zheng et al.)
Source: https://www.chinesestandard.net/PDF.aspx/GBT21510-2008