Liquid chromatographic determination of fluorescent derivatives of six sulfonamides in bovine serum and milk.
A rapid and sensitive liquid chromatographic (LC) method with fluorometric detection was developed to detect sulfadiazine, sulfathiazole, sulfamethazine, sulfamonomethoxine, sulfamethoxazole, and sulfadimethoxine residues in bovine serum and milk. p-Aminobenzoic acid (PABA) was added as an internal standard. The sulfonamides were extracted from samples and derivatized with fluorescamine, and 50 microL was injected into a NovaPak C18 LC column and eluted with acetonitrile-10 mM potassium phosphate (30 + 70, v/v). The sulfonamides were detected fluorometrically (excitation, 390 nm; emission, 475 nm), and their retention times ranged from 6.2 to 16.5 min without interference from coextractives. The detection limit for standard sulfonamide solution was 0.1 ng/mL; the calibration curves were linear between 1 and 100 ng/mL in the presence of PABA as internal standard. Recovery rates of sulfonamides from spiked samples (1 and 10 ppb) were 95.4-107.2 and 81.4-89.6% for serum and 80.7-91.1 and 62.6-84.1% for milk, respectively.
Tsai
C E
CE
Miyazaki University, Faculty of Agriculture, Department of Veterinary Public Health, Japan.
Kondo
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F
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D005450:Fluorescamine; D005470:Fluorometry; D008892:Milk / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000032:analysis* / Q000097:blood
D013449:Sulfonamides; D005450:Fluorescamine