GB/T 27827-2011 Test method for sister chromatid exchange in vivo of chemicals GB/T27827-2011 |tt||Standard compression package decompression password: www.bzxz.net
This standard specifies the terms and definitions, test principles, test methods, test data and reports for the test method for sister chromatid exchange in vivo of chemicals. This standard is applicable to the test for sister chromatid exchange in vivo of chemicals. This standard
was drafted in accordance with the rules given in GB/T1.1-2009.
This standard
is consistent with the technical content of OPPTS870. 5915:1998 "Sister Chromatid Exchange Test in Vivo" (English version) of the Office of Prevention, Pesticides and Toxic Substances (OPPTS) of the United States Environmental Protection Agency (USEPA).
This standard has been modified in the following structural and editorial aspects: ———
A chapter on scope has been added;
———The "Introduction" section in the original text has been used as the "Introduction" in this standard;
———Part of the content in the "Definition" in the original text has been used as "2 Terms and Definitions" in this standard;
———The measurement units have been uniformly changed to the legal measurement units of China.
This standard was proposed and managed by the National Technical Committee for Standardization of Hazardous Chemicals Management (SAC/TC251). || tt||
The drafting organizations of this standard are: Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Liaoning Provincial Institute of Occupational Disease Prevention and Control, China Chemical Economic and Technological Development Center, and China Institute of Inspection and Quarantine.
The main drafters of this standard are: Qu Bo, Lin Zheng, Li Xuefei, Bai Yu, Wang Xiaobing, and Chen Huiming.

ICS 13.300; F1. 100www.bzxz.net
National Standard of the People's Republic of China
GB/T 27827-—2011
Chemicals
In vivo sister chromatid exchange
Test method
Chemicals--Test method of in vivo sister chromatid exchangeIssued on 2011-12-30
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of ChinaStandardization Administration of Standardization of the People's Republic of China
Implementation on 2012-08-01
This standard was drafted according to the rules given in GB/T 1.1:2009. GB/T 27827—2011
This standard is consistent with the technical content of OPPTS8705915:1998 Sister Chromatid Exchange Test in Vivo (English version) of the Office of Prevention, Pesticides and Toxic Substances (OPPTS) of the United States Environmental Protection Agency (USEPA). This standard has made the following structural and editorial changes: adding a chapter on scope;
- The "Introduction" section in the original text is used as the "Introduction" in this standard;- Part of the content in the "Definition" in the original text is used as the "Terms and Definitions" in this standard;- The measurement units are unified-changed to the legal measurement units of my country. This standard is proposed and managed by the National Technical Committee for Hazardous Chemicals Management Standardization (SAC/TC251). Drafting units of this standard: Occupational Health and Poisoning Control Institute of China Center for Disease Control and Prevention, Liaoning Provincial Institute of Occupational Disease Prevention and Control, China Chemical Economic and Technical Development Center, China Institute of Inspection and Quarantine. The main drafters of this standard are Qu Bo, Lin Zheng, Li Xuefei, Bai Yu, Wang Xiaobing, Chen Hui, I
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GB/T27827--2011
This standard refers to USEPA0PPTS870.5915:1998 in vivo sister chromatid exchange test (English version), and the technical content is consistent with it. This test guide is a U.S. Environmental Protection Agency (EPA) Prevention, Pesticide and Toxic Substances Prevention Office (the One of the series of test guidelines issued by the Office of Pollution Prevention and Toxic Substances (OPPTS), the test objects are pesticides and toxic substances; the toxicological data obtained are submitted to the EPA for review in accordance with relevant regulations. The test guide issued by OPPTS is obtained by merging and weighing multiple test guidelines [including the guidelines issued by the Office of Pollution Prevention and Toxics (OPPTS), the guidelines in Chapter I, Subchapter R of the Code under Title 10 of the U.S. Code of Federal Regulations (Title 40, Chapter I, Subchapter R of the Code af Federal Regulations, CFR), the (Office of Pesticide Programs (the The OPPTS test guideline was developed by the United States Environmental Protection Agency (EPA) and the Organization for Economic Cooperation and Development (OECD). The purpose of combining multiple test guides to form a single set of OPPTS test guides is to meet the requirements of the U.S. Environmental Protection Agency for test data and minimize the differences in test methods. Applicability: This guideline was developed to meet the requirements of the Federal Insecticide, Fungicide, and Firefungicide Act (FIFRA) and the Toxic Substances Control Act (TSCA).
Background information: The references for the development of this synchronized OPPTS test guide include: OPPTS40 CFR 798.5915 in vivo sister chromatid exchange test and OPP84-2 mutagenicity test (Pesticide Evaluation Guide, Part F of Hazard Evaluation, Humans and Families) U.S. Department of Health and Welfare Report 540/09-82-025, 1982.
Sister chromatid exchange test detects the ability of chemicals to increase the exchange of DNA between two sister chromatids of chromosomes during replication. It can be tested in vitro using cultured mammalian cells or in vivo using non-dairy animals or mammalian tissues. The most commonly used are bone marrow cells and lymphocytes of mammals (such as mice, rats, hamsters), and human lymphocytes can also be used. TTTKANTKACA
1 Scope
Sister chromatid exchange in vivo for chemicals
Test method
GB/T27827—2011
This standard specifies the terms and definitions, test principles, test methods, test data and reports for the test method for sister chromatid exchange in vivo for chemicals.
This standard applies to the sister chromatid exchange test in vivo for chemicals. 2 Terms and definitions
The following terms and definitions apply to this document. 2.1
sisterchramatidexchange.scE The interphase exchange of two chromatids within a single chromosome during the replication phase of cell division. This exchange can be observed during the metaphase of the cell cycle and may require the cleavage, transfer and ligation of the DNA double helix. 3 Experimental Principle
The test substance is given to the animal through an appropriate route and then bromodeoxyuridine (BrdU) is given. Before the animal is euthanized, it is treated with the spondyl inhibitor colchicine to block cell division in metaphase. After the animal is euthanized, the test tissue is removed, the test tissue specimen is prepared, and the SCE is counted and analyzed under a microscope after staining. 4 Experimental Methods
4.1 Animal Selection
4.1.1 Species
Any suitable mammalian species can be used. The rodents used are mice, rats and hamsters. 4.1.2 Age
The animals should be healthy and newly grown.
4.1.3 Sex and number
Each treated group and control group should have at least 10 animals, 5 males and 5 females. If animals of the same sex or different numbers are used, appropriate reasons should be provided.
4.1.4 Grouping
The experimental animals should be randomly divided into the treated group and the control group. TTTKONYKAA
GB/T27827—2011
4.2 Control group
4.2.1 Control
Both positive control and negative (solvent) control should be set up. 4.2.2 Positive control
Chemicals known to cause SCE positive in in vivo tests should be used as positive controls. 4.3 Test substances
4.3.1 Solvent
The test substances should be dissolved in isotonic solution or hot water as much as possible. Poorly soluble test substances should be dissolved or suspended in a suitable solvent. The solvent should neither interfere with the test substance nor produce toxic effects. The test substance should be freshly prepared before use. 4.3.2 Dose level
The pre-test dose should be the maximum tolerated dose or the dose that causes toxic reactions (including death) or target cell toxicity in animals. LDs are more appropriate indicators. Its dose half-water can also be used. To determine the dose-response relationship, at least 3 dose half-water should be used. 4.3.3 Administration route
Generally, administration is by intraperitoneal injection or oral gavage. Other appropriate administration routes can also be used. 4.3.4 Number of administrations
Generally, administration is times. Repeated administration can also be carried out based on toxicity data. 4.4 Operation method
4.4.1 Exposure
Choose the exposure route according to the purpose of the test or the nature of the test sample. It is recommended to use intraperitoneal injection or oral gastric administration. 4. 4. 2 Administration of 5-Brau After the animal is exposed, 5-Brau can be administered by intraperitoneal injection, continuous infusion in the tail vein, subcutaneous injection, etc.
4.4.3 Treatment with spindle inhibitors
2 hours before the animal is killed, intraperitoneal injection of colchicine (<4mg/kg) is given). 4.4. 4 Preparation of bone marrow cell chromosome specimens After the animal is killed, the bone marrow is removed and the bone marrow cell suspension is prepared. Through the processes of hypotonicity, pre-fixation, fixation, and slide preparation, the bone marrow cell chromosome specimens are routinely prepared.
4.4.5 Staining method
The staining method can be determined according to the experimental plan. Fluorescent Sonochromatography is recommended. 2
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4.4.6 Counting cells
GB/T 27827—2011
The number of cells counted for each animal should be determined based on the number of animals used, the negative control SCE rate, the sensitivity and the selected statistical test level. Each slide should be numbered before microscopic examination. 5 Experimental data and report
5.1 Data processing
The results should be presented in a table: the number of SCEs in each metaphase cell and the number of SCEs in each metaphase cell chromosome should be listed. Before comparing the control group and the control group, the intra-group differences should be taken into account. 5.2 Statistical analysis
The statistical analysis should be performed using appropriate statistical methods. 5.3 Evaluation of the results of the experiment
5.3.1 The criteria for positive results are: a statistically significant increase in the number of SCEs showing a dose-effect relationship, or a repeatable and statistically significant increase in SCEs at at least one test point. 5.3.2 A test substance that has neither a statistically significant increase in the number of SCEs showing a dose-effect relationship nor a repeatable and statistically significant increase in SCEs at at least one test point can be considered to be unable to induce DNA fragment recombination under the test conditions. 5.3.3 The evaluation should consider both biological significance and statistical significance. 5.3.4 The results of the low-level test indicated that under the test conditions, the test substance could induce sister chromatid exchange in the bone marrow cells of the test animal; the low-level test results indicated that under the test conditions, the test substance could not induce sister chromatid exchange in the bone marrow cells of the test animal. 5.4 Experimental Report
The experimental report should include the following contents:
Animal strain, age, weight, number, sex;...-Test substance solvent, dose level, basis for dose selection, toxicity data, positive and negative controls;-Route and time of administration of test substance and 5-BrdU;-Name of spindle blocker, concentration and action time;-Time of animal sacrifice after administration of 5-BrdU;-Detailed information on slide preparation
-Standard for SCE counting;
-Dose-response relationship (if possible).
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References
[i] Allen, J. W, et al. for in vivo analysis af sister rhrornatid cxchangesusing 5-bromodeoxyuridine tablets. Cytogenerics Celf Genetics,1977,18:231-23?[3] I.att, SA tt al. Sister chromatid exchanges; A report of the US EPA Gene-Tox rogramMutatianResearch,198l,&7:17-62
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