Liquid chromatographic determination of multiple sulfonamide residues in bovine milk: collaborative study. 1994

M D Smedley
U.S. Food and Drug Administration, Center for Veterinary Medicine, Division of Residue Chemistry, Beltsville, MD 20705.

A collaborative study involving 8 laboratories was conducted on the determination of 8 sulfonamide residues in raw bovine milk using a liquid chromatographic (LC) method. The sulfonamides are extracted with chloroform-acetone, the organic phase is evaporated, the residues are dissolved in an aqueous potassium phosphate solution, and the fatty residues are removed by washing with hexane. The aqueous layer is collected, filtered, and injected onto an LC system, and the analyte is detected by ultraviolet (UV) absorption at 265 nm. To quantitate all 8 sulfonamides isocratically, 2 chromatographic conditions are required: 12% methanol in the mobile phase for 5 sulfonamides, and 30% methanol in the mobile phase for 4 sulfonamides. Sulfamethazine (SMZ), the most widely used sulfonamide, is detected by both systems. Collaborators were instructed to analyze 3 replicates each of control milk and control milk fortified at 3 levels. They were also provided with 20 blind incurred samples (10 samples in duplicate) to analyze. For 10 ppb fortified milk, the average interlaboratory recovery for the 8 sulfonamides ranged from 56.2% for sulfaquinoxaline (SQX) to 82.7% for SMZ in the 12% methanol mobile phase (SMZ12). Also at this level, Sr ranged from 3.2 for SQX to 8.9 for SMZ12, and SR ranged from 6.9 for sulfadimethoxine to 17.2 for SMZ in the 30% methanol system (SMZ30). At 10 ppb, RSDr and RSDR ranged from 5.7% for SQX to 10.8% for SMZ12, and 10.1% for sulfamerazine to 20.9% for SMZ30, respectively. These results demonstrate that the method is suitable for the determination of the 8 sulfonamide residues in milk at 10 ppb. However, the identification of positives by this procedure needs additional confirmation by procedures comparable to the specificity achievable by liquid or gas chromatography combined with mass spectrometry.

UI MeSH Term Description Entries
D008892 Milk The off-white liquid secreted by the mammary glands of humans and other mammals. It contains proteins, sugar, lipids, vitamins, and minerals. Cow Milk,Cow's Milk,Milk, Cow,Milk, Cow's
D002725 Chloroform A commonly used laboratory solvent. It was previously used as an anesthetic, but was banned from use in the U.S. due to its suspected carcinogenicity. Trichloromethane
D002853 Chromatography, Liquid Chromatographic techniques in which the mobile phase is a liquid. Liquid Chromatography
D004350 Drug Residues Drugs and their metabolites which are found in the edible tissues and milk of animals after their medication with specific drugs. This term can also apply to drugs found in adipose tissue of humans after drug treatment. Drug Residue,Residue, Drug,Residues, Drug
D005506 Food Contamination The presence in food of harmful, unpalatable, or otherwise objectionable foreign substances, e.g. chemicals, microorganisms or diluents, before, during, or after processing or storage. Food Adulteration,Adulteration, Food,Adulterations, Food,Contamination, Food,Contaminations, Food,Food Adulterations,Food Contaminations
D000096 Acetone A colorless liquid used as a solvent and an antiseptic. It is one of the ketone bodies produced during ketoacidosis.
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013418 Sulfamethazine A sulfanilamide anti-infective agent. It has a spectrum of antimicrobial action similar to other sulfonamides. Benzenesulfonamide, 4-amino-N-(4,6-dimethyl-2-pyrimidinyl)-,Sulfadimidine,Sulphamethazine,Sulfadimezine,Sulphamezathine
D013449 Sulfonamides A group of compounds that contain the structure SO2NH2. Sulfonamide,Sulfonamide Mixture,Sulfonamide Mixtures,Mixture, Sulfonamide,Mixtures, Sulfonamide

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