Codeofchina.com is in charge of this English translation. In case of any doubt about the English translation, the Chinese original shall be considered authoritative.
This standard is developed in accordance with the rules given in GB/T1.1-2009.
Evaluation method for efficacy of antibacterial and bacteriostasis
1 Scope
This standard specifies the selection principle and application of the evaluation method for antibacterial and bacteriostasis.
This standard is applicable to the identification of the efficacy of antibacterial and bacteriostasis of products with antibacterial and/or bacteriostatic functions.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.
GB 15979 Hygienic Standard for disposable sanitary products
FZ/T 73023 Antibacterial knitwear
Technical standard for disinfection (Edition 2002) issued by [the Ministry of Health of the People's Republic of China (WEIFAJIANFA [2002] No.282)]
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
antibacterial
process of using chemical or physical methods to kill bacteria or hinder their growth and reproduction, which can reduce their quantity and activity
3.2
bacteriostasis
process of using chemical or physical methods to inhibit or hinder the growth, reproduction and activity of bacteria
3.3
sustained antibacterial action
function where disinfection products with antibacterial action are able to kill or hinder the growth and reproduction of bacteria after being applied on the surface of an object for more than 7 d
3.4
neutralizer
reagent, in the test of killing microorganisms, used to eliminate the disinfectant left in the suspension for testing microorganisms and disinfectants and on the surface of microorganisms, so as to make these disinfectants lose their inhibitory and killing effects on microorganisms
4 Principles for selection of evaluation methods
4.1 Difference between antibacterial test and bacteriostatic test
Antibacterial test: To determine the antibacterial action of antibacterial products on bacteria and fungi, and neutralizer is needed to terminate the bactericidal action in the test.
Bacteriostatic test: To determine the bacteriostatic action of bacteriostatic products on bacteria and fungi, and neutralizer is not needed to terminate the bacteriostatic action in the test.
4.2 Selection of appropriate evaluation method according to product performance
4.2.1 Selection principle of evaluation method for bacteriostatic efficacy
As for bacteriostatic products, the method to evaluate their bacteriostatic efficacy shall be selected. Suspension quantitative bacteriostatic test shall be selected for liquid bacteriostatic products, the carrier immersion quantitative bacteriostatic test for paste or semisolid gel or viscous bacteriostatic products, the carrier bacteriostatic test for wet wipes or other products with bacteriostatic components, the bacteriostatic annulus test for soap and other solid bacteriostatic products, the immersion bacteriostatic test for the textile with soluble bacteriostatic components, and the retention bacteriostatic test for the bacteriostatic lotion with sustained bacteriostatic action.
4.2.2 Selection principle of evaluation method for antibacterial efficacy
As for antibacterial products, the method to evaluate their antibacterial efficacy shall be selected. Suspension quantitative bactericidal test shall be selected for liquid antibacterial products, the carrier immersion quantitative bactericidal test for gel or paste antibacterial products, the carrier bactericidal test for wet wipes or other products with antibacterial components, and the immersion antibacterial test for the textile with soluble antibacterial components.
The shake flask test shall be adopted for textiles, non-woven fabrics, fibers, etc., with insoluble antibacterial components, provided that they have been identified as not containing soluble antibacterial components.
The film sticking test shall be adopted for tiles, plastics, metals, coatings, etc. with insoluble antibacterial components; the continuous antibacterial test shall be adopted for products coated on the surface and having sustained antibacterial action after drying.
5 Evaluation method
5.1 Bacteriostatic efficacy
5.1.1 Suspension quantitative bactericidal test
5.1.1.1 Application scope
It is applicable to the determination of bacteriostatic efficacy of liquid bacteriostatic preparations such as sanitary lotion and bacteriostatic spray on microorganisms.
5.1.1.2 Reagents, culture media and apparatus
5.1.1.2.1 Test organisms
Staphylococcus aureus (ATCC 6538), Escherichia coli (8099), Candida albicans (ATCC 10231) and other organisms used based on specific uses of bacteriostatic agents.
5.1.1.2.2 Reagents
Diluent: 0.03 mol/L phosphate buffer solution (PBS) (pH 7.2 ~ 7.4); culture media: nutrient agar medium shall be used for the culture of Staphylococcus aureus and Escherichia coli, and Sabouraud Dextrose Agar (SDA) shall be used for the culture of Candida albicans.
5.1.1.2.3 Apparatus
Thermostatic water bath, timer, Class II biosafety cabinet, etc.
5.1.1.3 Test procedures
Take fresh slant culture obtained by culturing the test bacteria for 24 h, wash it with PBS, and then dilute it with PBS into a bacterial suspension of about 5.0×105 CFU/mL ~ 4.5×106 CFU/mL for later use. Take a sterile test tube, add 5.0 mL of the sample (using either stock solution or diluent according to the operating instructions), put it in the water bath at 20 ℃ ± 1 ℃ for 5 min, then add 0.1 mL of the test bacteria suspension, mix well quickly and start timing immediately. After the test bacteria interacts with the sample for the specified time in the instructions, pipette 1.0 mL of the mixed solution of the test bacteria and the sample to inoculate two plates respectively, into which the culture media shall be poured. If the number of colonies is not countable, PBS shall be used for ten-fold serial dilution, from which an appropriate dilution shall be selected, then pipette 1.0 mL of such dilution to inoculate two plates so as to culture and count the viable bacteria. At the same time, use PBS instead of the sample to carry out a parallel test as positive control. The number of colonies recovered from positive control shall be ranged from 1.0×104 CFU/mL ~ 9.0×104 CFU/mL. Take the PBS and the culture medium of the same batch as negative control. Culture all the test samples and control samples at 36 ℃ ± 1 ℃, for 48 h for gonites and for 72 h for Candida albicans respectively, and then observe the final results. Repeat the test for 3 times, and calculate the bacteriostasis rate respectively.
Foreword i
1 Scope
2 Normative references
3 Terms and definitions
4 Principles for selection of evaluation methods
5 Evaluation method
Codeofchina.com is in charge of this English translation. In case of any doubt about the English translation, the Chinese original shall be considered authoritative.
This standard is developed in accordance with the rules given in GB/T1.1-2009.
Evaluation method for efficacy of antibacterial and bacteriostasis
1 Scope
This standard specifies the selection principle and application of the evaluation method for antibacterial and bacteriostasis.
This standard is applicable to the identification of the efficacy of antibacterial and bacteriostasis of products with antibacterial and/or bacteriostatic functions.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.
GB 15979 Hygienic Standard for disposable sanitary products
FZ/T 73023 Antibacterial knitwear
Technical standard for disinfection (Edition 2002) issued by [the Ministry of Health of the People's Republic of China (WEIFAJIANFA [2002] No.282)]
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
antibacterial
process of using chemical or physical methods to kill bacteria or hinder their growth and reproduction, which can reduce their quantity and activity
3.2
bacteriostasis
process of using chemical or physical methods to inhibit or hinder the growth, reproduction and activity of bacteria
3.3
sustained antibacterial action
function where disinfection products with antibacterial action are able to kill or hinder the growth and reproduction of bacteria after being applied on the surface of an object for more than 7 d
3.4
neutralizer
reagent, in the test of killing microorganisms, used to eliminate the disinfectant left in the suspension for testing microorganisms and disinfectants and on the surface of microorganisms, so as to make these disinfectants lose their inhibitory and killing effects on microorganisms
4 Principles for selection of evaluation methods
4.1 Difference between antibacterial test and bacteriostatic test
Antibacterial test: To determine the antibacterial action of antibacterial products on bacteria and fungi, and neutralizer is needed to terminate the bactericidal action in the test.
Bacteriostatic test: To determine the bacteriostatic action of bacteriostatic products on bacteria and fungi, and neutralizer is not needed to terminate the bacteriostatic action in the test.
4.2 Selection of appropriate evaluation method according to product performance
4.2.1 Selection principle of evaluation method for bacteriostatic efficacy
As for bacteriostatic products, the method to evaluate their bacteriostatic efficacy shall be selected. Suspension quantitative bacteriostatic test shall be selected for liquid bacteriostatic products, the carrier immersion quantitative bacteriostatic test for paste or semisolid gel or viscous bacteriostatic products, the carrier bacteriostatic test for wet wipes or other products with bacteriostatic components, the bacteriostatic annulus test for soap and other solid bacteriostatic products, the immersion bacteriostatic test for the textile with soluble bacteriostatic components, and the retention bacteriostatic test for the bacteriostatic lotion with sustained bacteriostatic action.
4.2.2 Selection principle of evaluation method for antibacterial efficacy
As for antibacterial products, the method to evaluate their antibacterial efficacy shall be selected. Suspension quantitative bactericidal test shall be selected for liquid antibacterial products, the carrier immersion quantitative bactericidal test for gel or paste antibacterial products, the carrier bactericidal test for wet wipes or other products with antibacterial components, and the immersion antibacterial test for the textile with soluble antibacterial components.
The shake flask test shall be adopted for textiles, non-woven fabrics, fibers, etc., with insoluble antibacterial components, provided that they have been identified as not containing soluble antibacterial components.
The film sticking test shall be adopted for tiles, plastics, metals, coatings, etc. with insoluble antibacterial components; the continuous antibacterial test shall be adopted for products coated on the surface and having sustained antibacterial action after drying.
5 Evaluation method
5.1 Bacteriostatic efficacy
5.1.1 Suspension quantitative bactericidal test
5.1.1.1 Application scope
It is applicable to the determination of bacteriostatic efficacy of liquid bacteriostatic preparations such as sanitary lotion and bacteriostatic spray on microorganisms.
5.1.1.2 Reagents, culture media and apparatus
5.1.1.2.1 Test organisms
Staphylococcus aureus (ATCC 6538), Escherichia coli (8099), Candida albicans (ATCC 10231) and other organisms used based on specific uses of bacteriostatic agents.
5.1.1.2.2 Reagents
Diluent: 0.03 mol/L phosphate buffer solution (PBS) (pH 7.2 ~ 7.4); culture media: nutrient agar medium shall be used for the culture of Staphylococcus aureus and Escherichia coli, and Sabouraud Dextrose Agar (SDA) shall be used for the culture of Candida albicans.
5.1.1.2.3 Apparatus
Thermostatic water bath, timer, Class II biosafety cabinet, etc.
5.1.1.3 Test procedures
Take fresh slant culture obtained by culturing the test bacteria for 24 h, wash it with PBS, and then dilute it with PBS into a bacterial suspension of about 5.0×105 CFU/mL ~ 4.5×106 CFU/mL for later use. Take a sterile test tube, add 5.0 mL of the sample (using either stock solution or diluent according to the operating instructions), put it in the water bath at 20 ℃ ± 1 ℃ for 5 min, then add 0.1 mL of the test bacteria suspension, mix well quickly and start timing immediately. After the test bacteria interacts with the sample for the specified time in the instructions, pipette 1.0 mL of the mixed solution of the test bacteria and the sample to inoculate two plates respectively, into which the culture media shall be poured. If the number of colonies is not countable, PBS shall be used for ten-fold serial dilution, from which an appropriate dilution shall be selected, then pipette 1.0 mL of such dilution to inoculate two plates so as to culture and count the viable bacteria. At the same time, use PBS instead of the sample to carry out a parallel test as positive control. The number of colonies recovered from positive control shall be ranged from 1.0×104 CFU/mL ~ 9.0×104 CFU/mL. Take the PBS and the culture medium of the same batch as negative control. Culture all the test samples and control samples at 36 ℃ ± 1 ℃, for 48 h for gonites and for 72 h for Candida albicans respectively, and then observe the final results. Repeat the test for 3 times, and calculate the bacteriostasis rate respectively.
Contents of WS/T 650-2019
Foreword i
1 Scope
2 Normative references
3 Terms and definitions
4 Principles for selection of evaluation methods
5 Evaluation method
