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GB/T 15670.22-2017: Toxicological test methods for pesticides registration -- Part 22: DNA damage and repair/unscheduled DNA synthesis test in mammalian cells in vitro
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Toxicological test methods for pesticides registration -- Part 22: DNA damage and repair/unscheduled DNA synthesis test in mammalian cells in vitro
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GB/T 15670.22-2017
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Basic data | Standard ID | GB/T 15670.22-2017 (GB/T15670.22-2017) | | Description (Translated English) | Toxicological test methods for pesticides registration -- Part 22: DNA damage and repair/unscheduled DNA synthesis test in mammalian cells in vitro | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B17 | | Classification of International Standard | 65.100 | | Word Count Estimation | 9,963 | | Date of Issue | 2017-07-12 | | Date of Implementation | 2018-02-01 | | Issuing agency(ies) | General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China | | Summary | This standard specifies the basic principles, methods and requirements for DNA damage and repair / programmed DNA synthesis in mammalian cells in vitro. This standard is applicable to DNA damage and repair / programmed DNA synthesis of mammalian cells in vitro for pesticide registration. | GB/T 15670.22-2017: Toxicological test methods for pesticides registration -- Part 22: DNA damage and repair/unscheduled DNA synthesis test in mammalian cells in vitro
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Toxicological test methods for pesticides registration. Part 22. DNA damage and repair/unscheduled DNA synthesis test in mammalian cells in vitro
ICS 65.100
B17
National Standards of People's Republic of China
Pesticide registration Toxicology test methods
Part 22. In vitro mammalian cell DNA damage
And repair/extra-program DNA synthesis test
Part 22.DNAdamageandrepair/unscheduledDNAsynthesistest
2017-07-12 Posted
2018-02-01 implementation
General Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China
China National Standardization Administration released
Foreword
GB/T 15670 "pesticide registration toxicology test method" is divided into the following sections.
--- Part 1. General principles;
--- Part 2. Acute oral toxicity test Horn's method;
--- Part 3. Acute oral toxicity test sequential method;
--- Part 4. Acute oral toxicity test probability unit method;
--- Part 5. Acute dermal toxicity test;
--- Part 6. Acute inhalation toxicity test;
--- Part 7. Skin irritation/corrosivity test;
--- Part 8. Acute eye irritation/corrosiveness test;
--- Part 9. Skin allergy (sensitization) test;
--- Part 10. Short-term repeated oral toxicity (28 days) toxicity test;
--- Part 11. Short-term repeated transdermal (28 days) toxicity test;
--- Part 12. Short-term repeated inhalation exposure (28 days) toxicity test;
--- Part 13. Subchronic toxicity test;
--- Part 14. bacterial recovery mutation test;
--- Part 15. In vivo mammalian bone marrow polychromatic erythrocyte micronucleus test;
--- Part 16. In vivo mammalian bone marrow cell chromosome aberration test;
--- Part 17. Mammalian spermatogonial/spermatocyte chromosome aberration test;
--- Part 18. Rodent dominant lethality test;
--- Part 19. In vitro mammalian chromosome aberration test;
--- Part 20. In vitro mammalian cell gene mutation test;
--- Part 21. In vivo mammalian hepatocyte extracellular DNA synthesis (UDS) test;
--- Part 22. DNA damage and repair in vitro mammalian cells/DNA synthesis test outside the program;
--- Part 23. Teratogenicity test;
--- Part 24. Two generations of reproductive toxicity test;
--- Part 25. Acute delayed neurotoxicity test;
--- Part 26. Chronic toxicity test;
--- Part 27. Carcinogenicity test;
--- Part 28. Chronic toxicity and carcinogenicity combined test;
--- Part 29. Metabolism and toxicokinetic tests.
This section GB/T 15670 Part 22.
This section drafted in accordance with GB/T 1.1-2009 given rules.
This part is proposed and managed by the Ministry of Agriculture of the People's Republic of China.
This part of the drafting unit. Ministry of Agriculture pesticide test.
The main drafters of this section. Zhang Hongwei, Zhang Liying, Tao Chuanjiang.
Pesticide registration Toxicology test methods
Part 22. In vitro mammalian cell DNA damage
And repair/extra-program DNA synthesis test
1 Scope
GB/T 15670 provisions of this part of the in vitro mammalian DNA damage and repair/extra-procedural DNA synthesis test basic
Principles, methods and requirements.
This section applies to the registration of pesticides in vitro mammalian cells DNA damage and repair/extra-procedural DNA synthesis test.
2 Terms and definitions
The following terms and definitions apply to this document.
2.1
Out-of-procedure DNA synthesis unscheduledDNAsynthesis; UDS
Occurs in the S phase of the cell cycle to retain DNA synthesis beyond DNA synthesis.
2.2
Net nuclear silver netnucleargrain; NNG
Quantitative determination of intracellular UDS activity in autoradiographic UDS assays was calculated as the number of nuclei (NG) minus the area of the nucleus
Average cytoplasmic silver (CG), ie NNG = NG-CG. The number of NNGs is calculated from each cell, followed by one culture or parallel culture
The cells in the NNG are aggregated.
3 test purposes
The test system is based on the detection of the level of DNA repair synthesis to evaluate whether the test substance on the original mammalian cells and has been established
Of mammalian cell lines cause damage and damage to DNA. As UDS reflects the injury resection and repair process, and reflects the loss
The extent of the injury, it can be used as short-term chemical carcinogens to determine the end of biological tests.
4 Test Overview
The isolated and cultured non-S mammalian cells were transferred to a culture flask containing thymidine (3H-TdR) and a test substance and incubated
After a certain period of time, observed 3H-TdR incorporation, if the test substance caused cell DNA damage, will inevitably lead to cell autonomous DNA
Repair, integration of 3H-TdR into the DNA strand during repair, resulting in the incorporation of 3H-TdR into the repaired DNA. Reuse radioactive self
Development or liquid scintillation counting method to observe the incorporation of 3H-TdR amount, more incorporation, indicating that the test substance damage to DNA more widely.
Unless primary hepatocytes are used as target cells, cultured mammalian cells are treated with and without exogenous metabolic activation system
Under the situation with the test substance role.
In view of the primary culture of liver cells exist easily isolated culture, retention of liver microsomes and other advantages, it is recommended to use primary cultured hepatocytes as a test
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