GB/T 15670.3-2017 PDF EnglishUS$165.00 · In stock · Download in 9 seconds
GB/T 15670.3-2017: Toxicological test methods for pesticides registration - Part 3: Acute oral toxicity test - Up-and-down-procedure Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid
Similar standardsGB/T 15670.3-2017: Toxicological test methods for pesticides registration - Part 3: Acute oral toxicity test - Up-and-down-procedure---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT15670.3-2017 GB NATIONAL STANDARD OF THE PEOPLE'S REPUBLIC OF CHINA ICS 65.100 B 17 Toxicological test methods for pesticides registration - Part 3.Acute oral toxicity test - Up-and-down-procedure Issued on: JULY 12, 2017 Implemented on: FEBRUARY 1, 2018 Issued by. General Administration of Quality Supervision, Inspection and Quarantine of PRC; Standardization Administration of PRC. Table of ContentsForeword... 3 1 Scope... 5 2 Normative references... 5 3 Terms and definitions... 5 4 Purpose of the test... 6 5 Experimental overview... 6 6 Test methods... 6 7 Test results and evaluation... 10 8 Test report... 11 Appendix A (Normative) Table of dose levels (mg/kg body weight) with a starting dose of 175 mg/kg body weight and a slope of 1~8... 12 Appendix B (Normative) Calculation of LD50 in formal tests... 13 Toxicological test methods for pesticides registration - Part 3.Acute oral toxicity test - Up-and-down-procedure1 ScopeThis part of GB/T 15670 specifies the basic principles, methods and requirements for the up-and-down-procedure of acute oral toxicity testing. This part applies to acute oral toxicity tests conducted for pesticide registration.2 Normative referencesThe following documents are essential for the application of this document. For any dated referenced document, only the dated version applies to this document. For any undated referenced document, the latest version (including all amendments) applies to this document. GB 14925 Laboratory animal - Environment and housing facilities Regulations on Pesticide Registration Information (Order No. 10 of the Ministry of Agriculture of the People's Republic of China [2007])3 Terms and definitionsThe following terms and definitions apply to this document. 3.1 acute oral toxicity The health-damaging effects that occur in animals in the short term after the test substance is orally administered to experimental animals once or multiple times within 24 hours. 3.2 dose The amount of the test substance administered, which is expressed as the mass of the test substance received per unit body weight of the experimental animal (mg/kg body weight). 3.3 median lethal dose LD50 The statistical dose of a toxicant that causes half of the total number of animals to die after the test substance is given to the experimental animals once or multiple times within 24 hours. It is expressed as the mass of the test substance received by the experimental animals per unit body weight (mg/kg body weight). 3.4 dose-response relationship The relationship between the dose and the incidence of a specific effect.4 Purpose of the testAcute oral toxicity test is the first step to evaluate the toxicity of the test substance. Through short-term administration of toxicant by oral, the toxicity characteristics and dose-response relationship of the test substance are preliminarily understood, which provides a basis for acute toxicity classification, label management and other toxicology test dose selection.5 Experimental overviewThe up-and-down procedure (UDP) is a step-by-step administration procedure. Animals of the same sex are tested, and one animal is administered at a time. The first animal is administered a dose that is preferably lower than the estimated LD50.The increase or decrease in the subsequent dose of administration depends on the administration result (survival or death) of the previous animal. When the preliminary estimate of the LD50 of the test substance and the slope of the dose-response curve are not available, the computer simulation result suggests that the starting dose can be 175 mg/kg body weight, and the dose interval is calculated using the antilogarithm of 0.5 (corresponding to a default dose progression factor of 3.2). The point estimate value and 95% confidence interval of LD50 are calculated by the maximum likelihood method. NOTE. The dose progression factor is determined by the antilogarithm of the reciprocal of the slope of the dose-response curve (3.2 is the progression factor for a slope of 2). The number of animals used in the formal test is about 6~9.The number of animals used in the limit test is no more than 5.This method is only applicable to test substances that cause death within 1~2 days after administration, and is not applicable to test substances that are expected to cause death to happen more than 5 days (including 5 days) after administration.6 Test methods6.1 Test substances 6.4.1.2 If 3 of the 5 administered animals die within 48 hours or their death is within the 14-day observation period, the administration is terminated and the formal test begins. 6.4.2 Formal test 6.4.2.1 Before the experiment, the rats were fasted overnight but had unlimited access to water. 6.4.2.2 Selection of starting dose and dose progression factor. When an estimate of the lethal dose of the test substance is not available, the default starting dose is 175 mg/kg body weight. The dose sequence set corresponding to the progression factor 3.2 of a dose-response slope of 2 is 1.75 mg/kg body weight, 5.5 mg/kg body weight, 17.5 mg/kg body weight, 55 mg/kg body weight, 175 mg/kg body weight, 555 mg/kg body weight and 2000 mg/kg body weight; or 1.75 mg/kg body weight, 5.5 mg/kg body weight, 17.5 mg/kg body weight, 55 mg/kg body weight, 175 mg/kg body weight, 555 mg/kg body weight, 1750 mg/kg body weight and 5000 mg/kg body weight. If the slope of the dose-response curve of the test substance is known to be relatively steep or relatively flat, the dose progression factor can be reduced or increased. Appendix A lists the dose levels available with a starting dose of 175 mg/kg body weight and a slope of 1~8. 6.4.2.3 At the beginning of the test, weigh the fasted animals and administer the test substance orally using a gavage needle, one animal at a time, with an interval of 48 hours between each animal. The administration dose for the second animal depends on the result of the first animal's administration. If the animal dies or is in a dying state, the dose is adjusted down by one level according to the progression factor; if the animal survives, the dose is adjusted up by one level. When the subsequent doses show the opposite result to the starting dose, add 4 more animals to be administered the test substance sequentially and then terminate the test, or when any of the termination requirements (see 6.4.2.6) are reached, the test can be terminated. After administration, the rats shall continue to fast for 3 h~4 h. The observation period for each animal is 14 d. 6.4.2.4 If animals survive within 48 hours after administration but die later in the subsequent observation period, it indicates that the dose selection is inappropriate and consideration may be given to giving 2 more animals a dose lower than the dose that causes death, restarting the test and extending the observation period. 6.4.2.5 The test results are used to calculate the LD50 and 95% confidence interval of the test substance using the maximum likelihood method (see Appendix B). 6.4.2.6 The test can be terminated if any of the following conditions are met. a) When three animals survive the sequential administration at the upper dose range; b) When 5 opposite results occur among 6 animals randomly selected and sequentially administered; c) At least 4 animals are sequentially administered after the first adverse result and the likelihood ratio for each dose calculated starting from the first adverse result exceeds the critical value. If the LD50 and slope vary widely, the study can be terminated after 4~6 animals are administered sequentially after the opposite result has appeared. In some cases, the slope of the dose-response curve of the test substance is flat, and it may be necessary to add 15 animals. 6.5 Test observation and inspection 6.5.1 Clinical observation During the 24 hours after administration (especially within 30 minutes and the first 4 hours after administration) and the 14-day observation period, observe and record the poisoning reactions of each animal regularly every day. Clinical observations shall at least include. a) Central nervous system and neuromuscular system. abnormal posture, abnormal voice, restlessness, dullness, tremor, convulsion, paralysis, movement disorder, hypersensitivity or slowness in external response, abnormal behavior such as excessive and repeated scratching around the mouth, combing, circling, even self-mutilation, walking backward, etc.; b) Autonomic nervous system. pupil dilation or constriction, salivation and tearing; c) Respiratory system. nasal discharge, epistaxis, flaring of the nostrils, deep and slow breathing, tachypnea, wasp waist; d) Urogenital system. dirty perineum, discharge, vaginal or breast swelling; e) Skin and coat. skin congestion, cyanosis; fluffy or wet coat, dirty; f) Eyes. proptosis, conjunctival congestion, hemorrhagic discharge, corneal opacity; g) Digestive system. diarrhea, anorexia. The time and severity of tremors, convulsions, salivation, diarrhea, lethargy and coma, the death time or the recovery time shall be recorded. Animals on the verge of death or severe pain, as well as animals that survive the end of the experiment, shall be humanely8 Test reportThe test report shall at least include the following contents. a) Test name, test unit name and contact information, and report number; b) Test entrusting unit and contact information, sample acceptance date and sample sealing status; c) The start and end dates of the test, the person in charge of the test project, the technical person in charge of the test unit, and the date of issuance; d) Test summary; e) Name of the test substance, Chemical Abstracts Service Number (CAS Number) of the active ingredient (if known), code (if any), purity (or content), dosage form, production date (batch number), physical and chemical properties, solvents used for preparation and method; f) The species, strain, grade, number, weight, sex, source (supplier name, laboratory animal quality certificate number, laboratory animal production license number), quarantine and adaptation of the experimental animals, the feeding environment of the experimental animals, including temperature, relative humidity, feed, single cage or group feeding, and the license number of the experimental animal facility; g) Dose and group. including the principle or basis for selecting the dose, the method of animal grouping, sex and number of animals; h) Test conditions and methods. including main instruments and equipment, routes of administration, administration scheme, test period, observation indicators and method for calculating LD50, etc.; i) Test results. summarize item by item in the form of text description and table, including clinical manifestations of poisoning, onset, duration and recovery time of poisoning manifestations, time of death after administration, weight changes, correlation with dose, and gross anatomical and histopathological changes, etc., and indicate the statistical processing method of the results; j) Test conclusion. give clear conclusions, such as LD50, 95% confidence interval and toxicity classification, and discuss relevant issues when necessary; k) Description of the original record preservation status. termination regulations or terminate the test at a higher dose, so that the LD50 of the test substance is reported to exceed the upper limit and the hazard classification is made; b) The dose that causes animal death is higher than the dose that causes animal survival, and the LD50 is somewhere in between. This test will not give an accurate LD50 value (if the σ value is provided, the maximum likelihood ratio LD50 estimate can also be calculated); c) If the death and survival of the administered animals only occur at the same dose, all the animals die at a dose higher than this dose, and all the animals survive at a dose lower than this dose, then the LD50 is equivalent to this dose. B.1.4 Except for the above cases, the acute oral toxicity test application software package (AOT425StatPgm) can be used to perform the maximum likelihood function calculation. SAS (i.e. PROC NLIN), BMDP (i.e. program AR) or other computer program packages can also be used to complete the maximum likelihood function calculation. In the BASIC program, σ needs to be programmed as a parameter, and the output result is the estimated value of the logarithm (LD50) and its standard error. B.2 Confidence Interval (CI) calculation B.2.1 Only after the LD50 estimate of the formal test is obtained by using the AOT425StatPgm software package can the confidence interval be calculated and meaningful information be provided for the reliability and effectiveness of the formal test. A wide range of the LD50 confidence interval indicates that there is a lot of uncertainty in the reliability and effectiveness of the estimated LD50; when the confidence interval is narrow, the uncertainty of the LD50 is negligible, and the reliability and effectiveness are relatively high. When the formal test is repeated, the LD50 estimate obtained is closer to the original estimate, and both are closer to the true LD50 value. B.2.2 Two methods for estimating the interval estimate of the true LD50. a) At least three different doses of toxicity tests are carried out, and at least one animal survived and one animal died at the intermediate dose. The true LD50 and 95% confidence interval can be calculated using the maximum likelihood method. Due to the reduction in the number of experimental animals used, the confidence interval value is not very accurate. The provisions of random termination of the test have improved this to a certain extent, but there are still some differences from the true confidence interval. b) If all animals administered with a certain dose or a dose one level lower survive, and all animals administered with the next higher dose die, the confidence interval is between the two. It shall be considered an approximate ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of English version of GB/T 15670.3-2017 be delivered?Answer: The full copy PDF of English version of GB/T 15670.3-2017 can be downloaded in 9 seconds, and it will also be emailed to you in 9 seconds (double mechanisms to ensure the delivery reliably), with PDF-invoice.Question 2: Can I share the purchased PDF of GB/T 15670.3-2017_English with my colleagues?Answer: Yes. The purchased PDF of GB/T 15670.3-2017_English will be deemed to be sold to your employer/organization who actually paid for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. Our tax invoice, downloaded/delivered in 9 seconds, includes all tax/VAT and complies with 100+ countries' tax regulations (tax exempted in 100+ countries) -- See Avoidance of Double Taxation Agreements (DTAs): List of DTAs signed between Singapore and 100+ countriesQuestion 4: Do you accept my currency other than USD?Answer: Yes. www.ChineseStandard.us -- GB/T 15670.3-2017 -- Click this link and select your country/currency to pay, the exact amount in your currency will be printed on the invoice. Full PDF will also be downloaded/emailed in 9 seconds.How to buy and download a true PDF of English version of GB/T 15670.3-2017?A step-by-step guide to download PDF of GB/T 15670.3-2017_EnglishStep 1: Visit website https://www.ChineseStandard.net (Pay in USD), or https://www.ChineseStandard.us (Pay in any currencies such as Euro, KRW, JPY, AUD).Step 2: Search keyword "GB/T 15670.3-2017". Step 3: Click "Add to Cart". If multiple PDFs are required, repeat steps 2 and 3 to add up to 12 PDFs to cart. Step 4: Select payment option (Via payment agents Stripe or PayPal). Step 5: Customize Tax Invoice -- Fill up your email etc. Step 6: Click "Checkout". Step 7: Make payment by credit card, PayPal, Google Pay etc. After the payment is completed and in 9 seconds, you will receive 2 emails attached with the purchased PDFs and PDF-invoice, respectively. Step 8: Optional -- Go to download PDF. Step 9: Optional -- Click Open/Download PDF to download PDFs and invoice. See screenshots for above steps: Steps 1~3 Steps 4~6 Step 7 Step 8 Step 9 |
