液相色谱法测定牛血清和牛奶中六种磺胺类药物的荧光衍生物

Liquid chromatographic determination of fluorescent derivatives of six sulfonamides in bovine serum and milk.

作者信息

Tsai C E, Kondo F

机构信息

Miyazaki University, Faculty of Agriculture, Department of Veterinary Public Health, Japan.

出版信息

J AOAC Int. 1995 May-Jun;78(3):674-8.

PMID:7756881

Abstract

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.

摘要

建立了一种快速灵敏的带荧光检测的液相色谱（LC）方法，用于检测牛血清和牛奶中的磺胺嘧啶、磺胺噻唑、磺胺二甲嘧啶、磺胺间甲氧嘧啶、磺胺甲恶唑和磺胺二甲氧嘧啶残留。加入对氨基苯甲酸（PABA）作为内标。磺胺类药物从样品中提取并用荧光胺衍生化，取50微升注入NovaPak C18液相色谱柱，用乙腈-10 mM磷酸钾（30 + 70，v/v）洗脱。磺胺类药物用荧光法检测（激发波长390 nm；发射波长475 nm），其保留时间在6.2至16.5分钟之间，无共提取物干扰。标准磺胺类药物溶液的检测限为0.1 ng/mL；在以PABA作为内标的情况下，校准曲线在1至100 ng/mL之间呈线性。加标样品（1和10 ppb）中磺胺类药物在血清中的回收率分别为95.4 - 107.2%和81.4 - 89.6%，在牛奶中的回收率分别为80.7 - 91.1%和62.6 - 84.1%。