8. How is the adequacy of bedding materials to be handled?

This can be handled as are the analyses for possible contaminants in feed and water. The study director and associated scientists should consider the bedding and its

possible impact on the study. The results of this consideration should appear in the protocol.

9. What do the GLPs require in regard to assuring the genetic quality of animals used in a nonclinical laboratory study?

This is a scientific issue that is not specifically addressed by the GLPs. Suitability of the test system for use in a study is a protocol matter and any required testing procedure should be arrived at by the study scientists.

10. Do the GLPs require specific procedures for the microbiological monitoring of animals used in nonclinical laboratory studies?

The procedures used should be in accord with acceptable veterinary medical practice.

11. The Japanese are preparing animal care guidelines which are similar but no identical to the U.S. guidelines prepared by NIH. Would these be acceptable?

Japanese guidelines that are similar, but no less stringent, in the important particulars with the NIH guidelines would be acceptable to FDA.

12. What is the frequency of feed contaminant analysis?

If contaminant analyses are required by the protocol, then the GLPs require periodic analysis of the feed to ensure that the contaminant level is at or below that judged to be acceptable. Statistical procedures should be used to determine the frequency of analysis since this is dependent on the specific chemical characteristics of the interfering contaminant.

13. Is it necessary to use \interfering contaminants?

No. The methods should be appropriate for the analysis and FDA reserves the right to examine the raw data supporting the analytical results.

14. Do the GLPs require production facilities to be dedicated to the manufacture of specific animal feeds used in nonclinical laboratory studies? No.

15. Is a separate room required for animal necropsy?

No. The GLPs require separate areas and/or rooms as necessary to prevent any activity from having an adverse effect on the study. If the necropsy is done in an animal room, precautions should be taken to minimize disturbances that may interfere with the study.

SUBPART F

TEST AND CONTROL ARTICLES

Section 58.105 Test and control article characterization.

1. Is it necessary to retain samples of feed from nonclinical laboratory studies in which the feed serves as the control article?

Yes. It is not necessary, however, to retain reserve samples of feed from studies that involve test article administration by routes other than feed.

2. What expiration date is placed on the label of test articles whose stability is being assessed concurrently with the conduct of the study?

In this situation, the stability of the test article is unknown, but periodic analysis data exist. The label should contain a staetment such as \analysis results\be examined prior to continued use of the test article.

3. If analysis of the reserve samples is required by the Study Director or the QAU, is it permitted?

Yes, but sufficient reserve sample should be retained so that the sample is not exhausted.

4. Are physical and chemical tests conducted on test articles required to be done under the GLPs?

According to section 58.105, such tests conducted to characterize the specific batch of test articles used in the nonclinical laboratory study are covered.

Section 58.107 Test and control article handling.

1. With regard to safety studies in large animals (cattle, horses, etc.), must test article accountability be maintained and can the animals be used for food purposes?

Test article accountability must be maintained. For guidance on whether the treated animals can be used for food, you should contact the appropriate individuals in the Bureau of Veterinary Medicine.

Section 58.113 Mixtures of articles with carriers.

1. Do the GLPs require tests for homogeneity, concentration, and stability on mixtures of control articles used as positive controls? Yes.

2. Do test or control article concentration assays have to be performed on each batch of test or control article carrier mixture?

No. The GLPs require only periodic analysis of test or control article carrier mixtures.

3. What is the purpose of periodic analysis requirement for test or control article mixtures?

This requirement provides additional assurance that the test system is being exposed to protocol-specified quantities of test article. Whereas, in most instances proper assurance is obtained through adequate uniformity-of-mixing studies, adequate SOPs, and trained personnel, occasionally the mixing equipment can malfunction or other uncontrollable events can occur which lead to improper dosages. These events can be recognized through periodic analysis.

4. For acute studies, does the test article carrier mixture have to be analyzed (single dose studies)?

Yes, but the analysis need not be done prior to the study provided the mixture is stable in storage.

5. For liquid dosing studies where the test article mixture is made by dilution of the highest dose, which dose should be analyzed?

The lowest dose would be appropriate since it would confirm the efficacy of the dilution process, however the GLPs do not prohibit the analysis of any of the other doses.

6. Do homogeneity studies need to be done on solutions and suspensions of test articles used in acute nonclinical laboratory studies?

The answers to these questions are yes for suspensions of test articles and no for true solutions of test articles.

7. The analysis of test article mixtures that are used in acute studies is problematic. Usually at the stage of product development, the analytical method is not fully developed. Also, getting the analytical department to schedule the analysis is difficult. Stability is not a problem since fresh solutions are used. In view of the fact that acute studies are not pivotal in gaining approval of a research or marketing permit, is it necessary to analyze test article mixtures?

Yes. Although acute studies may be of lesser importance in assessing the safety of human drugs, they are important for animal drugs, biological products and certain food additives. For this reason, there must be some assurance that the test system was dosed with protocol specified quantities of test article. The GLPs do not require that the analysis be done prior to the use of the test article mixture provided that the mixture is stable on storage.

SUBPART G

PROTOCOL FOR THE CONDUCT OF A NONCLINICAL LABORATORY STUDY

Section 58.120 Protocol.

1. What are the proposed starting and completion dates for a nonclinical laboratory study?

There is a good deal of confusion on these dates and proper interpretation impacts on several GLP areas. Accordingly, the following clarificaion is offered: At the time of protocol development, the study director is to propose to management the

approximate time frame of the study. Section 58.120(a)(4), therefore, requires that the protocol contain the proposed starting and completion date of the study. These dates are somewhat discretionary provided that they are identified in the protocol. Suitable identification can be the date of first dosing of the test system to the date of last dosing, the date of necropsy of the last animal of test, the date of receipt of the test system to the date of final histopathological examination, or any combination of these or any other logical starting and completion dates. After this, the protocol is signed by the study director and forwarded for approval to management. Management approves, if indicated, signs and dates and at this point the study becomes a regulated study is carried on the Master Schedule Sheet until the study director submits a signed and dated final report. Thus, for Master Schedule Sheet purposes, the starting date of the study is the date of protocol approval by

management and the completion date of the study is the date of signature of the final report by the study director. Neither of the foregooing timeframes need be used to