Chemicals(organophosphorus substances) - Test method of delayed neurotoxicity following acute exposure
Some standard content:
ICS 13. 300; 11. 100
National Standard of the People's Republic of China
GB/T21770---2008
Chemicals (organophosphorus substances)-Test method of delayed neurotoxicity following acute exposure
Published on 2008-05-12
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China Administration of Standardization of the People's Republic of China
Implemented on 2008-09-01
National Standard of the People's Republic of China
Test method of delayed neurotoxicity following acute exposure of chemicals (organophosphorus substances)
GB/T 21770—2008
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GB/T 21770—2008bzxz.net
This standard is equivalent to the Organization for Economic Cooperation and Development (OECD) Chemical Testing Guide No. 418 (1995) "Delayed Neurotoxicity Test for Acute Infection of Organophosphorus Compounds" (English version). This standard has made the following editorial changes:
. ...Added the standard part;
The measurement unit is changed to the legal measurement unit of my country; Except for the reference part of OECD.
This standard is proposed and managed by the National Technical Committee for Standardization of Hazardous Chemicals Management (SAC/TC251). The responsible drafting unit of this standard is: Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention. The participating drafting units of this standard are: Ningbo Entry-Exit Inspection and Quarantine Bureau, Chemical Registration Center of State Environmental Protection Administration, Chinese Academy of Environmental Sciences, Safety Evaluation Center of Shenyang Research Institute of Chemical Industry. The main drafters of this standard are: Li Tao, Sun Jinxiu, Long Zaihao, Li Tao, Ma Zhongchun, Chen Xiaoqing, Lin Zhenxing. Product Partner Network
GB/T 21770—2008
OECD Introduction
1. The OECD periodically revises its chemical testing guidelines based on scientific developments and animal welfare considerations. This updated guideline uses a revised method, including measurement of neuropathic target enzyme (NTE, formerly known as neurotransmitter esterase) to determine the effect at sufficiently high doses, and no longer requires 21 or 30-minute checks. Inhibition of NTE in the brain and spinal cord 24 to 48 hours after exposure correlates well with the clinical and morphological effects of delayed neurotoxicity seen 10 to 20 days later. The NTE test model has been shown to be effective for all organophosphatases known to cause delayed neuropathy in humans. Therefore, quantitative data on NTE inhibition significantly improve the ability to determine whether some of the sometimes ambiguous results seen in behavior or histopathology are indicative of the presence of a delayed neurotoxic agent. 2. This updated version of the Guide to Testing of Chemicals was derived from the work of an expert working group convened in February 1992 in Paris on systematic short-term - updated version is mainly based on the views of the special meeting of the ECD Working Group on the coordination of neurological and (delayed) neurological tests held in Washington, D.C. on March 30, 1990. 3. In the assessment and evaluation of the toxic potential of chemical substances, the presence of other toxicants such as certain organophosphorus compounds may cause avoidance estimates. In addition, negative results obtained in in vitro screening tests may not be sufficient evidence. 4. No further evaluation is required under this test, except that the selected member states have recommended changes to the recommendations for the chemicals that cause specific types of neurotoxicity. Chemicals that have been evaluated as candidates for use in technical guidelines have now been observed. However, quality has been obtained from in vitro studies.
The results obtained by observation (under normal circumstances are not conclusive results), then it is necessary to conduct a retrospective test. 1 Scope
Delayed neurotoxicity test method for acute exposure of chemicals (organophosphorus compounds)
GB/T 21770—2008
This standard specifies the scope, definition of terms and abbreviations, basic principles of the test, test methods, test data and reports for animal tests on delayed neurotoxicity of chemical organophosphorus compounds. This standard is applicable to the detection of delayed neurotoxic effects after acute exposure to organophosphorus compounds. 2 Terms and definitions, abbreviations 2.1 Terms and definitions The following terms and definitions are used in this standard: Delayed neuropathy refers to a group of neurological syndromes characterized by the inhibition of neurotoxicity in neuronal tissues by organophosphorus acids, including organophosphates without the presence of an organophosphothioate, phosphinothioate or phosphoramidite. 2.2 Abbreviations The following abbreviations are used in this standard.
AChECactt
NTE(neuropa
TOCP(tri-o-cr
3 Basic principles of the test
terase
ergetes
Rsphai
all the co-existence
den and peripheral nerve distal axonal disease, as well as neuraminic acid, thiophosphorus substances related to peach phosphoramide.
The test substance was orally administrated once to hens, but domestic hens were protected in advance to avoid acute cholangiocarcinoma. The animals were observed for 21 days, including behavioral abnormalities, ataxia and paralysis. Animals were randomly selected from each group for differential testing (usually 24 h after poisoning). and 48h), especially NTE detection. After 21 days of infection, the retained animals were killed and the nerve tissue was selected for histological examination. 4 Experimental methods
4.1 Experimental animals
4.2 Animal species
Healthy, newly grown, 8- to 12-month-old domestic egg-laying hens (Gallusgallus domesticated species) were used. The size, breed and strain of the hens usually used should meet the standards. The animals should be raised and raised under conditions where they can move around. 4. 2. 1. Animal breeding environment
The animal breeding cage or fence should be large enough to allow the hens to move freely and to facilitate observation of the animal's gait. When the breeding environment is artificially lit, it should be cycled in a 12-hour light/12-hour dark cycle. Provide appropriate feed and free drinking water. 4.2.2 Animal preparation
Use healthy, newly grown hens that have not been infected with viral diseases, have not received drug treatment, and have no gait abnormalities. 21770-2008
Randomly divide into control group and poisoning group. At least 5 days should be adapted to the test laboratory environment before the start of the test. 4.3 Infection route and test material preparation
4.3.1 Oral infection is usually used, such as stomach, capsule, etc. 4.3.2 If the test material is axillary, it can be undiluted or diluted with a suitable excipient, such as corn oil: if the test material is solid, it may not be effectively absorbed in large doses in the form of gel, so it should be used as much as possible. It may be dissolved in the excipient. If the excipient used is not water, its toxicity characteristics should be understood. If it is not clear, its toxicity should be determined before the experiment. 4.4 Number of animals and grouping
4.4.1 Animal grouping includes excipient control group, positive control group and poisoning group. 4.4.2 There should be enough animals in the poisoning group and excipient control group to ensure that there are 6 animals for biochemical stasis test (two time points, three animals each) and 6 animals survive for pathological examination at the 21-day observation time. 4.4.3 The vehicle control group is treated the same as the toxicity group except that the test substance is not given. 4.4.4 The positive control can be a synchronous control or a recent historical control. The positive control group toxicated with known degenerative neurotoxicants should include at least 10 animals (3 for biochemistry and 3 for pathology). The widely used delayed neurotoxicants have TOCP. The historical data of positive controls should be updated regularly. And when the testing laboratory changes some key elements of the test (such as strains, feeding, and maintenance conditions), new positive control data must be established. 4.5 Preliminary test
The preliminary test is used to determine the number of hens and dose levels to be used in the formal test. It can be carried out by using an appropriate number of animals and dose levels. Since the results of the acute test will be used to determine whether to conduct the 28 test, the goal of the preliminary test is to determine the maximum dose used in the formal test. In order to determine the dose of the formal test, it is necessary to design a dose that causes the death of some animals in the preliminary test. However, to prevent death from acute irradiation, protective drugs known not to interfere with delayed neurotoxicity, such as atropine, may be used. Historical data on hens or other physiological information may also help in dose selection. 4.6 Limit Tests
When no toxic effects are observed in the test dose of 2000 mg/5 g·d) using the procedures specified in this test method, or when data from analysis of structurally related compounds also indicate that no toxicity is produced, there is no need to conduct higher dose tests. When human exposure levels indicate that a higher dose test is necessary, a limit test should not be used. 4.7 Formal Tests
Based on the results of the pilot test and the limit dose of 2000 mB/(kg, d), the dose level of the test substance designed in the formal test should be as high as possible. The number of animals that survived the death should be sufficient for biochemical (6) and pathological (6) tests. Atropine or other stimulants known not to disturb recurrent neurotoxicity should be used to prevent death from acute myokinetic effects. 4.8 Observation
Animals should be observed immediately after exposure. Careful observation should be made several times a day for the first two days and at least once a day thereafter, or until the animals are sacrificed as planned. All signs of toxicity should be recorded, including the time, type, severity and duration of clinical behavioral abnormalities. Ataxia can be assessed using a rating scale consisting of at least 4 levels, and paralysis should be recorded. Animals selected for pathological examination should be released from their cages at least twice a day and forced to perform escape activities, such as climbing stairs, to help observe the animals for mild toxic effects. All moribund animals should be removed and sacrificed for gross necrosis. 4.9 Body Observation
All hens should be weighed immediately before exposure and at least once a week thereafter. 4.10 Biochemistry
4.10.1 Six hens were randomly selected from each treated group and vehicle control group (when a simultaneous positive control was used) and three hens were randomly selected from the positive control group on a few days after exposure. The brain and lumbar spinal cord were removed and the NTE activity was measured. In addition, it is also useful to prepare and measure the NTE activity of the sciatic nerve tissue. Usually, three hens were taken from the control group and each treated group and killed 24 hours after exposure, and three hens were taken from the positive control group and killed 24 hours later. If the time of clinical toxicity symptoms observed (often assessed by observing the time of cholinergic symptoms) indicates that the toxicant has a very slow toxic effect in vivo, then the time of two samplings of nerve tissue should preferably be selected between 24 hours and 72 hours after exposure. GB/T 21770—2008
4.10.2 If deemed appropriate, the above-mentioned neural tissues may also be analyzed for AchE activity. However, AchE may reactivate in vivo, which may lead to an underestimation of the ability of the test substance to inhibit AchE. 4.11 Pathology
4.11.1 Gross anatomy
All animals (including those killed and sacrificed according to the plan) shall be subjected to gross anatomy, and the appearance of the brain and spinal cord shall be observed. 4.11.2 Histopathology
The neural tissues of animals shall be mainly examined under the microscope. The neural tissues shall be obtained from animals that survived the observation period and were not used for biochemical tests. The tissues shall be fixed in situ using perfusion techniques. The tissue sections must include the cerebellum (middle longitudinal hemisphere), cerebellum, spinal cord and peripheral nerves. Spinal cord sections include the upper cervical, mid-thoracic and lumbosacral regions: Peripheral nerve sections include the sciatic nerve, vena cava and its distal branches that innervate the cervical muscles. Sections should be stained with appropriate myelin and axon specific staining methods: 5 Experimental data and reports
5.1 Data
Must include individual data. All data should be presented in a grid, listing the number of animals in each group at the start of the experiment, the number of animals with injuries, behavioral abnormalities or biochemical changes, and the type and severity of injuries or toxic effects, as well as the corresponding percentages. 5.2 Evaluation of results
5.2.1 The results of the experiment should be evaluated based on the clinical behavior, biochemical and histopathological changes observed in the stained and control groups, as well as the incidence, severity and relationship between other effects observed. 5.2.2 Enumeration data should be evaluated using appropriate and generally accepted statistical methods, which should be selected when designing the experiment.
5.3 Test report
The test report must include the following information:
5.3.1 Test substance
a) Physical properties (including isomers, purity and physicochemical properties); 1) Name and identification code.
5.3.2 Excipients
If the excipient used is not water, the reason for its use should be stated. 5.3.3 Experimental animals
Animal strains:
Number and age of animals;
Origin of animals, and breeding conditions, etc.;
Each animal was tested at the beginning of the test.
Test conditions
Detailed description of the preparation, stability and homogeneity of the test substance; Detailed description of the toxicity of the test substance;
Quality of raw materials and drinking water;
Justification for dose selection;
Detailed description of the toxicity of the test substance, including information on excipients, administration volume and physical state; f)
Detailed description of the type of protective drugs used and the administration process, 5.3.5 Conclusions
Physical data;
Toxic reactions at each dose level, including mortality;
GB/T 21770--2008
Nature, severity and duration (whether reversible) of clinical abnormalities; Detailed description of biochemical test methods and results; d)
e) Gross anatomical examination results;
f) Detailed description of all histopathological findings; g) Statistical treatment of the results.
5.3. 6 Discussion of results
5.3.7 Conclusion
5.3.8 Interpretation of results
If the test results (including biochemistry, histopathology and clinical abnormalities) are negative, then no further delayed neurological tests are usually required. Further evaluation is required for suspicious or inconclusive results. Copyright infringement must be investigated
Book number: 155066: 1-32178
GB/T 21770-2008
Product Partner Network httn:6 Limit Tests
When the test is conducted using the procedures specified in this test method, the test substance is exposed to a dose of not less than 2000 mg/5 g·d) and no toxic effects are observed, or when the data obtained from the analysis of structurally related compounds also indicate that no toxicity will occur, then there is no need to conduct higher dose tests. When the human exposure level seems to indicate that a higher dose test is necessary, the limit test cannot be used. 4.7 Formal Tests
Based on the results of the preliminary test and the limit dose of 2000 mB/(kg, d), the dose level of the test substance designed in the formal test should be as high as possible. The number of animals that survived the death should meet the needs of biochemical (6) and pathological (6) tests. Atropine or other conserving drugs known not to disturb recurrent neurotoxicity should be used to prevent death caused by acute myocardial effects. 4.8 Observation
The animals should be observed immediately after exposure. Careful observation should be made several times a day for the first 2 days, and at least once a day thereafter, or until the animals are sacrificed as planned. All signs of toxicity should be recorded, including the time, type, severity and duration of clinical behavioral abnormalities. Ataxia can be evaluated using a rating scale consisting of at least 4 levels, and paralysis of the animals should be recorded. Animals selected for pathological examination should be released from their cages at least twice a day and forced to escape, such as climbing stairs, to help observe the animals for mild toxic effects. All moribund animals should be removed and sacrificed for gross autopsy. 4.9 Body
All hens should be weighed immediately before exposure and at least once a week thereafter. 4.10 Biochemistry
4.10.1 On certain days after exposure, 6 hens were randomly selected from each exposure group and vehicle control group (when a simultaneous positive control was set up), and 3 hens were randomly selected from the positive control group, and sacrificed. The brain and lumbar spinal cord were removed and their NTE activity was measured. In addition, it is useful to prepare and measure NTE activity in sciatic nerve tissue. Usually, 24 hours after exposure, three hens are killed from the control group and each exposed group, and 3 more are killed at 481, and the three hens in the control group are killed at 24 hours. If the observed time of clinical toxicity symptoms (often assessed by observing the time of cholinergic symptoms) indicates that the toxic substance exerts its toxic effects very slowly in vivo, then it is more appropriate to select two sampling times of nerve tissue between 24 hours and 72 hours after exposure. GB/T 21770—2008
4.10.2 If considered appropriate, the above nerve tissue can also be analyzed for AchE activity. However, AchE may be reactivated in vivo, thereby underestimating the ability of the test substance to inhibit AchE. 4.11 Pathology
4.11.1 Gross dissection
All animals (including those killed and marinated according to the plan) should be dissected to observe the appearance of the brain and spinal cord. 4.11.2 Histopathology
The nervous tissue of the animals should be examined microscopically. The nervous tissue should be obtained from animals that survived the observation period and were not used for biochemical tests. The tissue should be fixed in situ using perfusion technology. The tissue sections must include the cerebellum (middle longitudinal hemisphere), spinal cord, spinal cord and peripheral nerves. Spinal cord sections include the upper cervical, mid-thoracic and lumbosacral sections: the peripheral nerve sections include the sciatic nerve, vena cava and the distal region of their branches that innervate the cervical muscles. Sections should be stained with appropriate myelin and axon-specific staining methods: 5 Experimental data and reports
5.1 Data
Must include individual data. All data should be presented in a grid, listing the number of animals in each group at the start of the experiment, the number of animals with injuries, behavioral abnormalities or biochemical changes, and the type and severity of the injury or toxic effect, as well as the corresponding percentages. 5.2 Evaluation of results
5.2.1 The results of the experiment should be evaluated based on the clinical behavior, biochemical and histopathological changes observed in the stained and control groups, as well as the incidence, severity and relationship between other effects observed. 5.2.2 Enumeration data should be evaluated using appropriate and generally accepted statistical methods, which should be selected when designing the experiment.
5.3 Test report
The test report must include the following information:
5.3.1 Test substance
a) Physical properties (including isomers, purity and physicochemical properties); 1) Name and identification code.
5.3.2 Excipients
If the excipient used is not water, the reason for its use should be stated. 5.3.3 Experimental animals
Animal strains:
Number and age of animals;
Origin of animals, and breeding conditions, etc.;
Each animal was tested at the beginning of the test.
Test conditions
Detailed description of the preparation, stability and homogeneity of the test substance; Detailed description of the toxicity of the test substance;
Quality of raw materials and drinking water;
Justification for dose selection;
Detailed description of the toxicity of the test substance, including information on excipients, administration volume and physical state; f)
Detailed description of the type of protective drugs used and the administration process, 5.3.5 Conclusions
Physical data;
Toxic reactions at each dose level, including mortality;
GB/T 21770--2008
Nature, severity and duration (whether reversible) of clinical abnormalities; Detailed description of biochemical test methods and results; d)
e) Gross anatomical examination results;
f) Detailed description of all histopathological findings; g) Statistical treatment of the results.
5.3. 6 Discussion of results
5.3.7 Conclusion
5.3.8 Interpretation of results
If the test results (including biochemistry, histopathology and clinical abnormalities) are negative, then no further delayed neurological tests are usually required. Further evaluation is required for suspicious or inconclusive results. Copyright infringement must be investigated
Book number: 155066: 1-32178
GB/T 21770-2008
Product Partner Network httn:6 Limit Tests
When the test is conducted using the procedures specified in this test method, the test substance is exposed to a dose of not less than 2000 mg/5 g·d) and no toxic effects are observed, or when the data obtained from the analysis of structurally related compounds also indicate that no toxicity will occur, then there is no need to conduct higher dose tests. When the human exposure level seems to indicate that a higher dose test is necessary, the limit test cannot be used. 4.7 Formal Tests
Based on the results of the preliminary test and the limit dose of 2000 mB/(kg, d), the dose level of the test substance designed in the formal test should be as high as possible. The number of animals that survived the death should meet the needs of biochemical (6) and pathological (6) tests. Atropine or other conserving drugs known not to disturb recurrent neurotoxicity should be used to prevent death caused by acute myocardial effects. 4.8 Observation
The animals should be observed immediately after exposure. Careful observation should be made several times a day for the first 2 days, and at least once a day thereafter, or until the animals are sacrificed as planned. All signs of toxicity should be recorded, including the time, type, severity and duration of clinical behavioral abnormalities. Ataxia can be evaluated using a rating scale consisting of at least 4 levels, and paralysis of the animals should be recorded. Animals selected for pathological examination should be released from their cages at least twice a day and forced to escape, such as climbing stairs, to help observe the animals for mild toxic effects. All moribund animals should be removed and sacrificed for gross autopsy. 4.9 Body
All hens should be weighed immediately before exposure and at least once a week thereafter. 4.10 Biochemistry
4.10.1 On certain days after exposure, 6 hens were randomly selected from each exposure group and vehicle control group (when a simultaneous positive control was set up), and 3 hens were randomly selected from the positive control group, and sacrificed. The brain and lumbar spinal cord were removed and their NTE activity was measured. In addition, it is useful to prepare and measure NTE activity in sciatic nerve tissue. Usually, 24 hours after exposure, three hens are killed from the control group and each exposed group, and 3 more are killed at 481, and the three hens in the control group are killed at 24 hours. If the observed time of clinical toxicity symptoms (often assessed by observing the time of cholinergic symptoms) indicates that the toxic substance exerts its toxic effects very slowly in vivo, then it is more appropriate to select two sampling times of nerve tissue between 24 hours and 72 hours after exposure. GB/T 21770—2008
4.10.2 If considered appropriate, the above nerve tissue can also be analyzed for AchE activity. However, AchE may be reactivated in vivo, thereby underestimating the ability of the test substance to inhibit AchE. 4.11 Pathology
4.11.1 Gross dissection
All animals (including those killed and marinated according to the plan) should be dissected to observe the appearance of the brain and spinal cord. 4.11.2 Histopathology
The nervous tissue of the animals should be examined microscopically. The nervous tissue should be obtained from animals that survived the observation period and were not used for biochemical tests. The tissue should be fixed in situ using perfusion technology. The tissue sections must include the cerebellum (middle longitudinal hemisphere), spinal cord, spinal cord and peripheral nerves. Spinal cord sections include the upper cervical, mid-thoracic and lumbosacral sections: the peripheral nerve sections include the sciatic nerve, vena cava and the distal region of their branches that innervate the cervical muscles. Sections should be stained with appropriate myelin and axon-specific staining methods: 5 Experimental data and reports
5.1 Data
Must include individual data. All data should be presented in a grid, listing the number of animals in each group at the start of the experiment, the number of animals with injuries, behavioral abnormalities or biochemical changes, and the type and severity of the injury or toxic effect, as well as the corresponding percentages. 5.2 Evaluation of results
5.2.1 The results of the experiment should be evaluated based on the clinical behavior, biochemical and histopathological changes observed in the stained and control groups, as well as the incidence, severity and relationship between other effects observed. 5.2.2 Enumeration data should be evaluated using appropriate and generally accepted statistical methods, which should be selected when designing the experiment.
5.3 Test report
The test report must include the following information:
5.3.1 Test substance
a) Physical properties (including isomers, purity and physicochemical properties); 1) Name and identification code.
5.3.2 Excipients
If the excipient used is not water, the reason for its use should be stated. 5.3.3 Experimental animals
Animal strains:
Number and age of animals;
Origin of animals, and breeding conditions, etc.;
Each animal was tested at the beginning of the test.
Test conditions
Detailed description of the preparation, stability and homogeneity of the test substance; Detailed description of the toxicity of the test substance;
Quality of raw materials and drinking water;
Justification for dose selection;
Detailed description of the toxicity of the test substance, including information on excipients, administration volume and physical state; f)
Detailed description of the type of protective drugs used and the administration process, 5.3.5 Conclusions
Physical data;
Toxic reactions at each dose level, including mortality;
GB/T 21770--2008
Nature, severity and duration (whether reversible) of clinical abnormalities; Detailed description of biochemical test methods and results; d)
e) Gross anatomical examination results;
f) Detailed description of all histopathological findings; g) Statistical treatment of the results.
5.3. 6 Discussion of results
5.3.7 Conclusion
5.3.8 Interpretation of results
If the test results (including biochemistry, histopathology and clinical abnormalities) are negative, then no further delayed neurological tests are usually required. Further evaluation is required for suspicious or inconclusive results. Copyright infringement must be investigated
Book number: 155066: 1-32178
GB/T 21770-2008
Product Partner Network httn:1 Gross dissection
All animals (including those killed and martyred according to the plan) should be dissected and the appearance of the brain and spinal cord should be observed. 4. 11. 2 Histopathology
The nervous tissue of the animals should be examined microscopically. The nervous tissue should be obtained from animals that survived the observation period and were not used for biochemical tests. The tissue should be fixed in situ using perfusion technology. The tissue sections must include the cerebellum (middle longitudinal hemisphere), spinal cord, spinal cord and peripheral nerves. Spinal cord sections include the upper cervical, mid-thoracic and lumbosacral sections: the peripheral nerve sections include the sciatic nerve, vena cava and the distal region of their branches that innervate the cervical muscles. Sections should be stained with appropriate myelin and axon-specific staining methods: 5 Experimental data and reports
5.1 Data
Must include individual data. All data should be presented in a grid, listing the number of animals in each group at the start of the experiment, the number of animals with injuries, behavioral abnormalities or biochemical changes, and the type and severity of the injury or toxic effect, as well as the corresponding percentages. 5.2 Evaluation of results
5.2.1 The results of the experiment should be evaluated based on the clinical behavior, biochemical and histopathological changes observed in the stained and control groups, as well as the incidence, severity and relationship between other effects observed. 5.2.2 Enumeration data should be evaluated using appropriate and generally accepted statistical methods, which should be selected when designing the experiment.
5.3 Test report
The test report must include the following information:
5.3.1 Test substance
a) Physical properties (including isomers, purity and physicochemical properties); 1) Name and identification code.
5.3.2 Excipients
If the excipient used is not water, the reason for its use should be stated. 5.3.3 Experimental animals
Animal strains:
Number and age of animals;
Origin of animals, and breeding conditions, etc.;
Each animal was tested at the beginning of the test.
Test conditions
Detailed description of the preparation, stability and homogeneity of the test substance; Detailed description of the toxicity of the test substance;
Quality of raw materials and drinking water;
Justification for dose selection;
Detailed description of the toxicity of the test substance, including information on excipients, administration volume and physical state; f)
Detailed description of the type of protective drugs used and the administration process, 5.3.5 Conclusions
Physical data;
Toxic reactions at each dose level, including mortality;
GB/T 21770--2008
Nature, severity and duration (whether reversible) of clinical abnormalities; Detailed description of biochemical test methods and results; d)
e) Gross anatomical examination results;
f) Detailed description of all histopathological findings; g) Statistical treatment of the results.
5.3. 6 Discussion of results
5.3.7 Conclusion
5.3.8 Interpretation of results
If the test results (including biochemistry, histopathology and clinical abnormalities) are negative, then no further delayed neurological tests are usually required. Further evaluation is required for suspicious or inconclusive results. Copyright infringement must be investigated
Book number: 155066: 1-32178
GB/T 21770-2008
Product Partner Network httn:1 Gross dissection
All animals (including those killed and martyred according to the plan) should be dissected and the appearance of the brain and spinal cord should be observed. 4. 11. 2 Histopathology
The nervous tissue of the animals should be examined microscopically. The nervous tissue should be obtained from animals that survived the observation period and were not used for biochemical tests. The tissue should be fixed in situ using perfusion technology. The tissue sections must include the cerebellum (middle longitudinal hemisphere), spinal cord, spinal cord and peripheral nerves. Spinal cord sections include the upper cervical, mid-thoracic and lumbosacral sections: the peripheral nerve sections include the sciatic nerve, vena cava and the distal region of their branches that innervate the cervical muscles. Sections should be stained with appropriate myelin and axon-specific staining methods: 5 Experimental data and reports
5.1 Data
Must include individual data. All data should be presented in a grid, listing the number of animals in each group at the start of the experiment, the number of animals with injuries, behavioral abnormalities or biochemical changes, and the type and severity of the injury or toxic effect, as well as the corresponding percentages. 5.2 Evaluation of results
5.2.1 The results of the experiment should be evaluated based on the clinical behavior, biochemical and histopathological changes observed in the stained and control groups, as well as the incidence, severity and relationship between other effects observed. 5.2.2 Enumeration data should be evaluated using appropriate and generally accepted statistical methods, which should be selected when designing the experiment.
5.3 Test report
The test report must include the following information:
5.3.1 Test substance
a) Physical properties (including isomers, purity and physicochemical properties); 1) Name and identification code.
5.3.2 Excipients
If the excipient used is not water, the reason for its use should be stated. 5.3.3 Experimental animals
Animal strains:
Number and age of animals;
Origin of animals, and breeding conditions, etc.;
Each animal was tested at the beginning of the test.
Test conditions
Detailed description of the preparation, stability and homogeneity of the test substance; Detailed description of the toxicity of the test substance;
Quality of raw materials and drinking water;
Justification for dose selection;
Detailed description of the toxicity of the test substance, including information on excipients, administration volume and physical state; f)
Detailed description of the type of protective drugs used and the administration process, 5.3.5 Conclusions
Physical data;
Toxic reactions at each dose level, including mortality;
GB/T 21770--2008
Nature, severity and duration (whether reversible) of clinical abnormalities; Detailed description of biochemical test methods and results; d)
e) Gross anatomical examination results;
f) Detailed description of all histopathological findings; g) Statistical treatment of the results.
5.3. 6 Discussion of results
5.3.7 Conclusion
5.3.8 Interpretation of results
If the test results (including biochemistry, histopathology and clinical abnormalities) are negative, then no further delayed neurological tests are usually required. Further evaluation is required for suspicious or inconclusive results. Copyright infringement must be investigated
Book number: 155066: 1-32178
GB/T 21770-2008
Product Partner Network httn:
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