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Determination and Confirmation of 5-hydroxyflunixin in Raw Bovine Milk Using Liquid Chromatography Tandem Mass Spectrometry

Determination and Confirmation of 5-hydroxyflunixin in Raw Bovine Milk Using Liquid Chromatography Tandem Mass Spectrometry

Pamela L Boner, Dave D W Liu, William F Feely, Matthew J Wisocky, Jinn Wu

J Agric Food Chem. 2003 Jun 18;51(13):3753-9.

PMID: 12797739

Abstract:

A method was developed and validated to determine 5-hydroxyflunixin in raw bovine milk using liquid chromatography tandem mass spectrometry (LC/MS/MS). The mean recovery and percentage coefficient of variation (%CV) of 35 determinations for 5-hydroxyflunixin was 101% (5% CV). The theoretical limit of detection was 0.2 ppb with a validated lower limit of quantitation of 1 ppb and an upper limit of 150 ppb. Accuracy, precision, linearity, specificity, ruggedness, and storage stability were demonstrated. A LC/MS/MS confirmatory method using the extraction steps of the determinative method was developed and validated for 5-hydroxyflunixin in milk from cattle. Briefly, the determinative and confirmatory methods were based on an initial solvent (acetone/ethyl acetate) precipitation/extraction of acidified whole milk. The solvent precipitation/extraction effectively removed incurred ((14)C) residues from milk samples. The organic extract was then purified by solid phase extraction (SPE) using a strong cation exchange cartridge (sulfonic acid). The final SPE-purified sample was analyzed using LC/MS/MS. The methods are rapid, sensitive, and selective and provide for the determination and confirmation of 5-hydroxyflunixin at the 1 and 2 ppb levels, respectively.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP75369618	5-Hydroxyflunixin		75369-61-8	Price

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