std5f_Nutrient_Specifications.docx

03/26/2014 NIH STD No. 5F

1. Scope:

1.1 This specification covers nutrient and chemical contaminant analyses on samples of laboratory animal diet and bedding purchased under NIH contracts.

2. REQUIREMENTS

2.1 Laboratory analyses to be performed.

Nutrient and Contaminant Analyses - Diet and bedding samples may be analyzed for all nutrients, required by the animal, and/or contaminants stipulated in the NIH specifications. The analyses will be performed in accordance with the most recent issue of Official Methods of Analyses of the Association of Official Analytical Chemists (A.O.A.C.) or other officially approved analytical methods as cited below. Methods of analyses will not be altered without a written request from the Contractor and written approval from the NIH Contracting Officer, Contract Officer Representative and Laboratory Animal Nutritionist.

Analyses Methodology

Samples shall be collected using sterile gloves and/or apparatus. Submitted diet and bedding samples shall be homogenized by the laboratory then the homogenized sub-samples shall be used for nutrient and contaminant analyses. The analyzed diet sample will be a representative sample of the diet sample submitted by the NIH. The results of each nutrient and contaminant assay will be validated using industry-wide acceptable quality assurance measures. Final approval for all quality assurance methods shall be made by the NIH COR.

Diet samples shall be subjected to the following analyses in accordance with the indicated procedure.

Nutrient Method Description

1. Total Protein 945.18* Kjeldahl; Dumas 968.06 992.15

2. Total Fat 920.39 Ether Extract

922.06 Acid Hydrolysis

954.02

3. Crude Fiber 962.09 Digestion: Gravimetric

4. Ash 923.03 Muffle Furnace

5. Moisture 934.01 Vacuum Oven;

925.09 Gravimetric

Linolenic Acids)

7. Calcium 975.03 Atomic Absorption 935.13

8. Phosphorus 964.06 Alkalimetric 2011.14 Method; OES

9. Potassium 975.03 AA Spectroscopy

2011.14 OES

10. Sodium 985.35 AA Spectroscopy

11. Magnesium 985.35 AA Spectroscopy

975.03 OES

12. Manganese 985.35 AA Spectroscopy

975.03 OES

13. Iron 985.35 AA Spectroscopy

975.03 OES

14. Zinc 985.35 AA Spectroscopy

975.03 OES

15. Copper 985.35 AA Spectroscopy

17. Iodine 934.02 Knapheide-Lamb Method

18. Chloride 943.01 Titrimetric Method

19. Vitamin A 974.29 Colorimetric

2012.09 HPLC

20. Vitamin D 982.29 HPLC

21. Vitamin E 971.30 HPLC; Column

2012.09 Chromatography

and Spectro- photometry

22. Thiamin 942.23 Spectrometry and

953.17 flurometry

23. Riboflavin 970.65 Fluorometric

981.15 Spectroscopy

940.33

24. Niacin 961.14 Colorimetric 944.13 981.16

25. Pantothenic Acid 945.74 Microbiological;

Turbidimetric Titrimetric

26. Choline 999.14 Colorimetric;

2012.18 HPLC, IEC

2012.20

27. Pyridoxine 961.15 Microbiological;

Chromatography, Spectroscopy

28. Total Folic Acid 944.12 Microbiological:

2004.05 Spectroscopy

29. Biotin JAOAC,49:882, Spectroscopy

(1996);JAFC, 23:1157,(1975)

30. Vitamin C 984.26 Fluorometric;

967.22 Spectroscopy

31. Vitamin B12 952.20 Microbiological

2011.08 HPLC

2011.09

32. Arginine 994.12 Amino acid Analyzer

35. Leucine 994.12 Amino acid Analyzer

Contaminant Method Description

1. Chlorinated 970.52 Gel Permeating Hydrocarbon 983.21 Chromatography Pesticides 984.21

2. Polychlorinated 983.21 GLC biphenyls (PCBs)

3. Organo-phosphate 968.24 GLC Pesticides 970.52 970.53 974.22

4. Malathion 968.24 GLC 970.52 970.53 974.22

5. Lead 972.25 Atomic Absorption 986.15 Spectrophotometry

6. Arsenic 986.15 Atomic Absorption 957.22 Spectrophotometry

7. Cadmium 986.15 Atomic Absorption

973.34 Spectrophotometry

8. Mercury 971.21 Atomic Absorption Spectrophotometry

9. Aflatoxins 970.43 TLC (B1, B2, G1, G2) 975.35 Romer minicolumn 975.36 977.16

10. Deoxynivalenol 970.43 TLC; GLC

986.17 Mass Spectrometry

986.18

11. Fumonisin(B1,B2,B3) 970.43 GLC

12. Zearalenone 994.01 Immunoassay

Contaminant (cont) Method Description

19. Nitrate 968.07 Colorimetric

20. Nitrosamines 991.28 GLC

969.46

22. Salmonella 967.25 Microbiological 967.26 989.12 989.14 989.15 991.13

23. E. Coli 991.14 Microbiological 992.30 983.25 990.11

25. Mold 984.29 Microbiological 995.21 997.02

26. Listeria 2003.12 PCR

27. Clostridium 974.38 Microbiological Perfringes

28. Isoflavones 2001.10 Chromatography

* The numbers indicate AOAC method

2.2 Completion Time for Assays- All specified nutrient and contaminant analyses shall be completed within 10 working days after the feed samples are received by the laboratory.

2.3 Laboratory Records- All data associated with the analyses herein specified shall be retained by the laboratory for at least two years. These data shall be made available to the NIH COR upon request. The laboratory shall maintain a log with a complete identification of the samples. The log shall include the following information: date each sample is received; product identification; manufacturing date; and, the assay results.

2.4 Reporting Results - Analyses shall be provided to the NIH Contract Officer Representative by email within 24 hours after all analyses are complete. Report shall include but not limited to, date of the report, sample ID, laboratory ID, and analytical results including units. The results are the property of the U. S. Government. The laboratory is not authorized to make results available to anyone but the NIH COR without written permission from the NIH COR.

2.5 Contractor's Restrictions - To be eligible for award of the contract the contractor shall have the capabilities to perform nutrient and contaminant analyses as specified herein. The Contractor shall provide documentation that all Subcontractors are capable of performing the analyses using approved analytical methods conforming to NIH standards. Any laboratory that is not independent of the NIH animal feed or bedding contractors shall not be eligible for award of a contract to perform these services.

3.0 Personnel

3.1 Each individual engaged in the conduct of or responsible for the supervision of feed and bedding analyses shall have education, training, and experience, or combination thereof, to enable that individual to perform the assigned functions. A current summary of training and experience and job description for each individual engaged in or supervising the conduct of the feed analyses shall be maintained at the testing facility. There shall be a sufficient number of personnel for the timely and proper conduct of the analyses. Personnel shall take necessary personal sanitation and health precautions designed to avoid contamination of feed and bedding. Personnel engaged in feed and bedding analyses shall wear clothing appropriate for the duties they perform. Such clothing shall be changed as often as necessary to prevent microbiological,and/or chemical contamination of feed and bedding samples.

3.2 The testing facility shall identify a scientist or other professional of appropriate education, training and experience or combination thereof as the director of feed and bedding analyses. The director has overall responsibility for the technical conduct of the analyses, documentation and reporting of results. The director shall assure that: (a) All results including unanticipated observations are accurately recorded and verified. (b) Unforeseen circumstances that may affect the quality and integrity of the analyses are noted and corrective action is taken and documented. (c) Analytical methods are as specified in the most current NIH Std. 5 or have been approved by the COR. (d) All good laboratory practices specified in the most current NIH Std 5 are followed.

4.0 Quality Assurance

4.1 A testing facility shall have a quality assurance unit which shall be responsible for monitoring feed and bedding analyses to assure management that the facilities, equipment, personnel, methods, practices, records and controls are in compliance with the specifications of this contract. The quality assurance unit shall be entirely separate from and independent of the personnel engaged in the direction and conduct of the feed and bedding analyses.

4.2 The quality assurance unit shall: (a) Maintain copies of all protocols pertaining to the methods required to analyze the feed and bedding. (b) Inspect each analytical procedure at intervals adequate to assure the integrity of the analyses and maintain written and properly signed records of each periodic inspection showing the date of inspection, the person performing the inspection, findings and problems, and action taken to resolve existing problems. (c) Submit to the director of feed and bedding analyses written status reports noting problems and corrective actions taken. (d) Determine that no deviation from approved analytical methods were made without proper authorization and documentation. (e) Prepare annual inspection reports for the NIH specifying the date the inspection was performed and findings.

4.3 The NIH COR or designated NIH representative shall have access to the written procedures established for the inspection and may request laboratory management to certify that inspections are being implemented, performed, documented and followed-up in accordance with this specification.

4.4 The laboratory shall participate in an official independent check-sample program (Such as the AAFCO and AOAC program).

5.0 Facility

5.1 The testing facility shall be of suitable size and construction to facilitate the proper conduct of feed and bedding analyses. As necessary to prevent contamination, there shall be separate areas for receipt, storage and analyses of feed and bedding.

5.2 The facility shall be kept in a sanitary condition at all times.

6.0 Laboratory Operation- Standard Operating Procedures (available pre-award)

6.1 The laboratory shall have standard operating procedures (SOP) in writing setting forth analytical methods that the NIH is satisfied are adequate to insure the quality and integrity of the data generated in the course of nutrient and contaminant analyses of the laboratory animal feed and bedding. All deviations in analytical methods from the SOPs shall be authorized by the director of laboratory animal feed and bedding analyses and shall be documented in the raw data. Significant changes in SOPs shall be properly authorized in writing by the director of feed and bedding analyses. The NIH COR shall be notified within 24 hours of any changes in SOPs. The NIH COR will make the final decision concerning any changes to analytical methodology.

6.2 SOPs shall be established for, but not limited to, the following:

(1) Receipt, identification, storage handling, and method of sampling of the laboratory animal feed and bedding.

(2) Laboratory tests.

(3) Data handling, storage, and retreival.

(4) Maintenance and calibration of equipment.

6.3 Each laboratory area shall have immediately available laboratory manuals and SOPs relative to the laboratory procedures being performed. Published literature may be used to supplement SOPs.

6.4 A historical file of SOPs, and all revisions thereof, including the dates of such revisions, shall be maintained. The Contractor shall submit SOPs to the COR upon request.

7.0 Equipment

7.1 Equipment used in the performance of the nutrient and contaminant analyses of laboratory animal feed and bedding and equipment used for facility environmental control shall be of appropriate design and adequate capacity to function according to the protocol. The equipment shall be suitably located for operation, inspection, cleaning and maintenance.

7.2 Maintenance and Calibration of Equipment

(c) Written records shall be maintained for all inspection, maintenance, testing, calibrating and/or standardizing operations. These records containing the date of the operation, shall describe whether the maintenance operations were routine and followed the written SOP. Written records shall be kept of non-routine repairs performed on equipment as a result of failure or malfunction. Such records shall document the nature of the defect, how and when the defect was discovered, and any remedial action taken in response to the defect.

8.0 Reagents and Solutions- All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer or concentration storage requirements, and expiration date. Deteriorated or outdated reagents and solutions shall not be used.

9.0 Verification of Results - The Government reserves the right to verify results of all analyses. This may be accomplished by inspection of data accumulated by the contractors, by submitting coded samples collected from products delivered to NIH, or by conducting the analyses on duplicate product samples at other laboratories.

10.0 Inspection- The testing facilities (Contractor and subcontractor) shall permit authorized employees of the National Institutes of Health, at reasonable times and in a reasonable manner to inspect the facility, all records(and in the case of records also to copy), and specimens required to be maintained regarding analyses within the scope of this contract. A pre-award inspection of all laboratory facilities to be used in providing these services will be made. The Government also reserves the right to make either announced or unannounced inspections at any time during the contract period.