Method Needs and Fitness for Purpose Statement

Date: September 19, 2004

Project: Determination of oxytetracycline hydrochloride in animal feeding stuffs

Project Leader: Richard Larson and Nancy Thiex

South Dakota State University

Brookings, SD

Telephone: 605-688-6171

Fax: 605-688-6295

Email: Richard.Larson@sdstate.edu

Project Team:

1.0 Needs:

Oxytetracycline hydrochloride is used as an aid in stimulating appetite, maintaining weight gains, and feed efficiency, in pigs, swine, beef cattle, chickens and turkeys. In addition it also serves as an aid in egg production, hatchability and eggshell quality during periods of stress and/or bacterial infections or potential infections in broiler, replacement and laying chickens, and turkeys. Oxytetracycline is used as an aid in the prevention, treatment and reduction of losses due to bacterial infections in lambs, pigs and calves. And, it is used as an aid to reduce the incidence of abortion in swine and bloat in young cattle. Oxytetracycline is used for the treatment of various diseases in salmon and trout.

Methodology is required to verify the levels of oxytetracycline in various feeds, supplements and premixes. Methodology is also required to determine contamination levels to verify clean out of manufacturing equipment for the prevention of cross contamination. Due to potential residues in tissues and animal products for use in food, withdrawal periods are required.

1.1 Performance Needs (based on laboratory sample)

Accuracy: (See Recovery)

Type A & Type B Medicated/Additive/Premix/Mineral Mix, 95 – 105%,

Type C Medicated Feed, 90 – 110 %

Contamination, > 80%

Applicability: For use in broiler, layer and replacement chicken, turkey, pig, swine, beef cattle, calf, calf milk replacer, lamb, salmon and trout feeds

Premixes: 110 – 440 g//kg

Medicated/Additive Complete Feed for broiler chickens: 55 - 220 mg/kg

Medicated/Additive Complete Feed for laying chickens: 55 - 220 mg/kg

Medicated/Additive Complete Feed for replacement chickens: 55 - 220 mg/kg

Medicated/Additive Complete Feed for turkeys: 55 - 220 mg/kg

Medicated/Additive Complete Feed for lambs: 22 - 110 mg/kg

Medicated/Additive Complete Feed for pigs/swine: 55 - 550 mg/kg

Medicated/Additive Complete Feed for calves: 55 mg/kg

Medicated/Additive Complete Feed for cattle: 20 - 100 mg/kg*

Medicated/Additive Complete Feed for fish: 0.50 – 30 g/kg

Premix: 110 – 440 g/kg

*Note: estimated from feed levels from usage of 75 mg/head/day and 500 mg/head/day range in cattle

Detection Limits:

Medicated/Additive/Premix/Mineral Mix 3 mg/kg

Contamination (non-fish feed): 1 mg/kg

Contamination (fish feed): x g/kg

Determination Limits:

Medicated/Additive/Premix/Mineral Mix 10 mg/kg

Contamination (non-fish feed): 3 mg/kg

Contamination (fish feed): X mg/kg

Precision Repeatability:

Medicated/Additive/Premix/Mineral Mix: CV_r = or < 5 %

Contamination: CV_r = or < 10 %.

Precision Reproducibility

Medicated/Additive/Premix/Mineral Mix: CV_R = or < 10 %

Contamination: CV_R = or < 20 %.

Range: 3 mg/kg to 440 g/kg

Recovery:

Medicated/Additive/Premixes/Mineral Mixes 95 – 105 %

Contamination > 80%

Selectivity:

The method is to be free of interferences from matrix, other drugs, vitamins, minerals. Oxytetracycline hydrochloride is compatible with the following drug/drug combinations: amprolium (broiler chickens, replacement chickens, turkeys), arsanilic acid (broiler chickens, replacement chickens, turkeys, swine), hygromycin B (broiler chickens, replacement laying chickens, swine), 3-nitro-4-hydroxyphenylarsonic acid (broiler chickens, replacement chickens, laying chickens, turkeys, swine), zoalene (broiler chickens, replacement chickens, turkeys), neomycin (beef cattle).

Linearity of standard curve: r ≥ 0.999, and 95% confidence limit of the y intercept includes zero.

Special Considerations:

Performance of this method should be comparable to or exceed plate assay AOACI OMA 968.50.

The method is to be rugged/robust and critical parameters are to be identified and controlled.

Method performance criteria are to be defined. Familiarization plan is to be suggested which will demonstrate that the laboratory analyst can capably perform the method prior to analyzing samples.

Quality control plan is to be suggested along with warning and out of control limits.

Traceability:

Standards and acceptable sources are to be identified. Standards are to provide activities and uncertainty.

Method Performance:

Fitness for Purpose Review

Fitness for Purpose Statement