Some standard content:
ICS 13. 300,11. 100
National Standard of the People's Republic of China
GB/T27823—2011
Chemicals-Test method of acute dermal toxicity-Fixed dose procedure2011-12-30 Issued
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of ChinaStandardization Administration of the People's Republic of China
2012-08-01Implementation
This standard was drafted in accordance with the rules given in GB/T1.1—2009. GB/T27823—2011
This standard is consistent with the technical content of the Organization for Economic Cooperation and Development (OECD) Chemical Test Method ND.434 (2004) "Acute Dermal Toxicity Fixed Dose Procedure (English Version)".
This standard has made the following structural and editorial changes: 1. Added a chapter on scope (see Chapter 1); --- The measurement units are changed to the legal measurement units of my country; --- The "Definition*" in Appendix 1 of OECD 434 is used as Chapter 2 of this standard; --- The "Pre-test Considerations" of OECD 434 is placed in the introduction of this standard. This standard is proposed and managed by the National Technical Committee for the Promotion of Hazardous Chemicals Management Standards (SAC/TC251). The drafting units of this standard are: Institute of Occupational Health and Poisoning Control, Chinese Center for Disease Control and Prevention, Tianjin Entry-Exit Inspection and Quarantine Bureau, China Chemical Economic and Technological Development Center, Jiangsu Entry-Exit Inspection and Quarantine Bureau. The main drafters of this standard are: Hou Fensui, Wang Hua, Zhang Yuan, Xu Bin, Yang Ting, Yue Zhuo, Liu Wei, Tang Lijun. 1
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In light of scientific developments and animal welfare requirements, OECD chemical testing guidelines need to be reviewed regularly. The acute dermal toxicity guideline TG402 was first adopted in 1987. After the adoption of the revised acute oral toxicity test fixed dose weekly method (FDP) (OECD 420[3]) and the abolition of OECD Guideline 401 in December 2001, the acute dermal fixed dose method (FDP) was proposed to be appropriate. This method mainly uses several fixed toxic doses, using single-sex Animals (usually female animals) are used to test acute percutaneous toxicity.
Traditional methods often use animal death as the only toxicity endpoint when evaluating acuteness. In 1984, the British Society of Toxicology proposed a new acute toxicity test method, which is to select a fixed dose for the test substance to be poisoned]. This method avoids using animal death as the observation endpoint: instead, it uses the appearance of obvious poisoning symptoms at a dose among several weekly fixed doses as the observation endpoint. This dose is also the basis for the toxicity classification of the test substance. This test also uses the above method. In order to achieve the OECD guidelines for observation points According to requirement 41, it is recommended to optimize the test method to minimize the suffering of animals and reduce the use of animals as much as possible. The statistical rationality of the FDP test has been evaluated by mathematical model-5.
This method can provide information on the hazardous properties of the test substance. For test substances that can cause acute effects, the toxicity of the test substance can be graded and classified according to the United Nations Global Harmonized Classification and Labeling of Chemicals (GHS) based on the results of this test 5). All information obtained on the test substance should be considered before the test, such as the name, chemical structure, physicochemical properties of the test substance, any other in vivo or in vitro toxicity test results, quantitative structure-activity relationship (QSAR) data, toxicological data of structurally similar chemicals, the intended use of the test substance, and the possible degree of human exposure to the test substance. Based on the above information, a suitable starting dose is selected for the test. TTTKAONYKACA
1 Scope
Acute dermal toxicity of chemicals
Fixed dose test method
GB/T 27823—2011
This standard specifies the terms, definitions and abbreviations, test principles, test methods, test data and reports for the fixed-dose test method for acute dermal toxicity of chemicals.
This standard applies to the fixed-dose method for testing the dermal toxicity of chemicals. 2 Terms, definitions and abbreviations
The following terms, definitions and abbreviations apply to this document: 2.1 Terms and definitions
Eacute dermal toxicity
Acute dermal toxicity
Adverse effects of a test substance produced by a single dermal exposure in a short period of time (within 24 hours). 2.1.2
Erident toxicity
Obvious toxicity after exposure to animals. If a higher fixed concentration is used, most animals are expected to show obvious pain, lethargy or even death.
Dose
The amount of the test substance administered. The amount of the test substance given per unit body weight of the test animal (e.g., mg/kg). 2.1.4
impending death
A sign of impending death or dying in an animal. Signs of impending death in rodents include withdrawal, lateral recumbency, recumbency, and trembling: 2.1.5
LD50
The dose of a substance that causes death in 50% of the animals after a single oral exposure. The LD value is expressed as the amount of the test substance per unit body weight of the test animal (mg/kg). Statistical derivatives.
limit dose
The upper limit dose of the test (2000 mg/kg or 5000 mg/kg). 2.1.7
noribnd status
Noribnd status
A state in which an animal is about to die or cannot survive despite treatment. 2.1.8
prelictable dealh
predictable death
clinical manifestations indicate that the animal will die at some time before the end of the test, for example, loss of water and food.
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2.2 Abbreviations
GHS: Globally Harmonize System of Classification and Labelling of Chemicals
Note: Jointly initiated by OECD for human health and environment, UN Committee of Experts on the Transport of Dangerous Goods for physical and chemical properties and ILO for hazard communication, and coordinated by the Inter-Organization Programme for the Sound Management of Chemicals (IOMC). 3 Experimental principles
Only moderately toxic doses should be used, avoiding doses that are expected to cause lethality. Similarly, doses that are expected to cause severe pain due to corrosive or severe irritant effects should be avoided. Animals that die, show obvious pain, or are in persistent pain should be euthanized and classified as dead for the purpose of interpreting the results. Criteria for killing dead and extremely distressed animals, as well as instructions for identifying moribund animals, can be found in separate guidelines. Groups of animals of a single sex are exposed to appropriate fixed doses listed in Appendices A and B in a stepwise manner. The starting dose is selected based on preliminary studies, that is, the dose that is expected to produce obvious toxic manifestations without causing severe toxic reactions or mortality. Clinical manifestations related to pain, suffering, and near-mortality are described in detail in the OECD Guidelines on Humane Endpoints [41]. The fixed dose is further increased or decreased based on the occurrence of sexual manifestations or mortality. The study is terminated when: obvious toxicity occurs or no more than one animal dies, or no toxic effects are observed at the highest dose, or animal mortality occurs at the lowest dose. 4 Test methods
4.1 Selection of animal species
4.1.1 Adult rats can be used.Rabbits or guinea pigs. Female animals are routinely recommended. If male animals are used, sufficient reasons for this choice should be provided. For the most appropriate sex selection, by investigating traditional oral and acute inhalation toxicity tests, it was found that there was generally almost no difference in sex sensitivity, and those cases where there was a difference showed that females were slightly more sensitive. There is no such data for dermal administration, but it can be inferred that the difference in sex sensitivity is similar to that of oral and inhalation routes. However, if the toxicological and toxicokinetic data of structural analogs indicate that females may be more sensitive, female animals should be used, but sufficient reasons for this choice should be provided in this case. 4.1.2 Choose animal strains commonly used in the laboratory + healthy, 8-12 weeks old, and females should be nulliparous and non-pregnant. The weight difference should be within ±20% of the average weight of the experimental animals. 4.2 Housing conditions
The temperature of the animal room should be maintained at 22 ± 3°C. The relative humidity should be at least 30%, preferably not more than 70% (except during cleaning), and the relative humidity should be between 50% and 60%. Artificial night-time, 12h light and dark intercourse, regular laboratory simple food, free drinking water. Animals should be kept in groups, but the number of animals per pet should not affect the effective observation of each animal. 4.3 Animal preparation
4.3.1 At least 5 days should be adapted to laboratory conditions before the start of the poisoning. Animals should be randomly selected and numbered. 4.3.2 About 21 hours before the test, cut or shave the hair on the back of the animal. The hair removal area should be at least 10% of the body surface area. The weight of the animal should be considered when determining the hair removal and coverage area.
4.4 Percutaneous poisoning
4.4.1 Apply the test substance evenly to a sample area not larger than 10% of the body surface area. For highly toxic test substances, the skin coating area can be smaller, but the thin layer applied should be as thin as possible and as evenly as possible to cover the entire poisoning area. During the 24-hour exposure period, use porous gauze2
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and non-microscopic adhesive tape to keep the test substance in contact with the skin. The affected area should also be covered in an appropriate manner to prevent the gauze from falling off and to ensure that the test substance is not licked by the animal. When the solid test substance (sometimes it needs to be ground) is exposed, it should be fully moistened with water or an appropriate solvent/excipient should be used to ensure good contact with the skin. When using solvents/excipients, the effect of solvents/excipients on the ability of the test substance to penetrate the skin should be considered. Axillary tests generally do not require dilution. 4.4.2 In order to prevent the test substance from being turned over and eaten by the animal, a brake can be used to restrict the animal's activities, but complete braking is not recommended. After the exposure, remove the residual test substance with water or other appropriate solvents. 4.5 Procedure
4.5.1 Preliminary study
4.5.1.1 The purpose of the preliminary study is to select an appropriate starting dose for the formal study. The test substance is administered to one animal at a time in a sequential manner according to the flow chart in Appendix A. The preliminary study can be stopped once the starting dose for the formal study can be determined (or death occurs at the lowest fixed dose).
4.5.1.2 The starting dose for the preliminary study is the dose that is expected to produce significant toxicity from the fixed doses of 50 mg/kg, 200 mg/kg, 1000 mg/kg and 2000 mg/kg. If possible, the starting dose can be selected based on in vivo and in vitro test data of the same chemical or structural analogues. If this information is not available, 1000 mg/kg is used as the starting dose. 4.5.1.3 After the end of the exposure of the first animal, at least 24 hours should be allowed between exposure of the second animal and exposure of the second animal. The observation period for all animals should be at least 14 days.
4.5.1.4 If the test animal dies or shows obvious toxicity at the lowest fixed dose (50 mg/kg) in the preliminary test, the test can be terminated and the substance is classified as a GHS acute toxicity Category 1 substance (hereinafter referred to as "Category 1") (see Appendix A). If further confirmation is required, the second animal should be exposed to 50 mg/kg. If this animal also dies, it is confirmed to be GHS Category 1 and the test should be terminated immediately. If the second animal survives, up to three other animals can be exposed to 50 mg/kg. The time between the exposure of the first and the second animals should be long enough to determine whether the first exposed animal is likely to survive. If the second of the three animals dies, the test should be terminated immediately and there is no need to expose the third animal. Appendix B lists the 50 mg/kg dose. Toxicity classification at a weekly dose of 100 mg/kg: the death of 2 or more animals is Category 1 (result A), and the death of one animal is GHS Category 2 (result B). 4.5.1.5 It is recommended to consider using an upper limit weekly dose of 5000 mg/kg only in a few cases (see note C). From the perspective of animal welfare, animal tests in the GHS Category 5 range (2000 mg/kg to 5000 mg/kg) are not encouraged and will only be considered when the results of this test are likely to be directly related to the protection of human or animal health and the protection of the environment. 4.5.2 Formal test
4.5.2.1 Number of animals and exposure dose
4.5,2.1.1 Appendix B indicates the next test steps starting from the starting dose. One of the following three steps is required: terminate the test and specify the appropriate hazard level, continue the test at a higher or lower fixed dose. However, in order to avoid unnecessary suffering to animals, the formal test will no longer repeat the dose that caused death in the preliminary test (see Appendix B). Experience has shown that at the starting dose, the most likely result is that the substance can be classified without further testing. If the test is conducted in a dose-reducing manner and 2 to 3 animals are observed to die (belonging to the category of result A), then, considering animal welfare, the test should be stopped and the test substance should be classified according to the result C of the next dose day.
4. 5.2.1.2 Five animals of the same sex are required for each dose to be tested, of which 10 are from the pre-test of the selected dose. 4.5.2.1.3 The interval between the exposure of animals at different doses depends on the onset, duration and severity of the toxicity indications. The exposure of animals at the next dose should be postponed until it can be confirmed that the animals in the previous dose group have survived. The interval between exposures of consecutive doses is 3 or 4 days to observe delayed toxicity. If the upper limit dose of 50 DD mg/kg is considered, the test should be carried out in accordance with Appendix C. 4.5.2.2 Limit test
Limit test is mainly carried out when there is evidence that the test substance is likely to be non-toxic, that is, it is only carried out when the dose exceeds the specified dose (4.5.2.2, 1
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toxic). The toxicity data of the test substance can come from the test data of structurally similar compounds and their mixtures, or the test data of the test substance product, the name and proportion of the toxic components in the test substance composition. If there is no or only a little relevant toxicity data, or if the test substance is predicted to be toxic, a formal test should be carried out. 4.5, 2.2.2 Use the standard test procedure, take one animal in the preliminary test and test it at a starting dose of 2,000 mg/kg (5,000 mg/kg in some cases), and take another 4 animals in the formal test to test at this dose level, which can be used as the limit test of this standard. In some cases, in order to meet the requirements of some regulatory agencies, it is necessary to follow the test procedure until the GHS Class 5 classification is stopped before the test (see Appendix C).
4.5.2.3 Observation
4.5.2.3.1 Animals should be observed at least once within 30 minutes of exposure and regularly during the first 24 hours, with particular attention paid to the response during the first 4 hours. Thereafter, they should be observed once a day for a total of 14 days, unless the animal dies within 14 days. However, the observation period is not fixed but depends on the nature of the toxic manifestations, the time of their onset and the length of their recovery period. The time of onset and disappearance of toxic manifestations is of great importance, especially since toxic manifestations can be delayed. All observations should be recorded systematically and a separate record should be made for each animal. Animals that are dying, in obvious pain or in severe distress should be euthanized immediately. The time when the animal is euthanized and the time when it is found dead should be recorded as accurately as possible.
4.5.2.3.2 Observations should include changes in the skin, coat, eyes, mucous membranes, respiratory system, circulatory system, autonomic and central nervous systems, and body movement and behavioral patterns. Particular attention should be paid to the occurrence of convulsions, convulsions, convulsions, drowsiness, stupor, sleep, and coma. Attention should be paid to the principles and standards summarized in the "Humane Endpoints Guidance Document" [1]. Animals that are moribund or showing obvious pain should be euthanized. Also, care should be taken (such as setting up a control group exposed to air) when examining the clinical manifestations of sound effects, not to mistake early changes in appearance and respiration caused by the exposure process for toxic reactions of the test substance. 4.5,2.4 Body weight
Weigh the animals at least once a week after the day of dosing. At the end of the experiment, weigh the surviving animals and euthanize them. 4.5.2.5 Pathology
Perform gross autopsy on all animals. Record the results of gross autopsy observations for each animal. For animals that survive more than 24 hours after the initial dose of poisoning, perform histopathological examinations on the organs that show changes in their gross anatomy to collect useful information. 5 Experimental data and reports
5.1 Data
Data for each animal should be provided, including the number of animals used in each group, the number of animals that showed toxic symptoms, the number of animals that died or were euthanized during the experiment, the time of death of each animal, the description of the toxic reaction and its time course, whether the toxic reaction is recoverable, and the pathological results. 5.2 Test report
If possible, the test report should include the following information: a) Tested animals:
1) Physical properties, purity, relevant physicochemical properties (including isomers); 2) Identification information, including the Chemical Abstracts Service (CAS) number (if available). b) Excipients (if any); explain the reason for using excipients, and explain the reason when the excipient is not water. c) Test animals:
1) Species and strain;
2) Animal purity level (if known); 3)
Number of animals, age and sex (if male, explain the reason for not choosing females); 4
1) Source, feeding conditions, feed, housing data, etc.; 5) If any animals died during the test, the date and time of death. d Test conditions
1) Detailed composition of the test substance: including the physical properties of the test substance used during the exposure; 2)
Detailed information on the test substance, including exposure volume and time; GB/T 27823—2011
Detailed information on the quality of feed and drinking water (including type/source of feed, source of drinking water); 3
Justification for the selection of the initial exposure dose;
5) Method of random animal selection
e) Results:
1) Report in a table the toxic manifestations and exposure dose of each animal (e.g., animal deaths, nature, severity and duration of toxic manifestations);
Weight of each animal on the day of exposure. Weight per week after exposure. Weight at the time of death and euthanasia, date and time of death of animals that died during the experiment;
Time course of toxic manifestations for each animal, and whether it recovered; 3)
4) If possible, the results of household examination and histopathological examination of each animal, 5.3 Discussion and interpretation of results,
5.4 Conclusion.
GB/T 27823—2011
Starting dose: 60nog/kg
1 animal
50mg/kg
GHS Category 1*
Formal industry
Starting dose
(mg/ke)
1 month animal
50mg/kg
GHS Category 1 Category*
Formal test
Starting agent blue,
(mg/kg)
Obvious toxicity
No recurrence
Appendix A
(Normative Appendix)
Preliminary test flow chart
Starting dose: 200mg/kg
1 animal
200amg/kg
1 animal
1000 ing/kg
1 other animal
1000mg/kg
1 animal
2000mng/kg
1 animal
2000mg/kg
+5Dmg/kg, for the result (), a supplementary procedure can also be selected to determine the GHS Classification - Safety See 4.5.I Preliminary test
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
Bright bean and wheat
Non-toxic
1 animal
200mz/kg
1 animal
200mg/kg
Initial dose: 100mg/kg
1 animal
1000me/kg
1 animal
1000mg/kg
AOI Preliminary test
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
Bright bean and wheat
Non-toxic
1 animal
200mz/kg
1 animal
200mg/kg
Initial dose: 100mg/kg
1 animal
1000me/kg
1 animal
1000mg/kg
AOI Preliminary test
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
1 animal
50mg/kg
GIIS Class 1*
Formal trial
(mg/kg)
Bright bean and wheat
Non-toxic
1 animal
200mz/kg
1 animal
200mg/kg
Initial dose: 100mg/kg
1 animal
1000me/kg
1 animal
1000mg/kg
AO
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