Simultaneous Determination of Chloramphenicol, Florfenicol, and Thiamphenicol Residues in Milk by Gas Chromatography with Electron Capture Detection

Abstract

A gas chromatographic (GC) method is described for determining residues of chloramphenicol (CAP), florfenicol (FF), and thiamphenicol (TAP) in raw milk, with meta-nitrochloramphenicol (mCAP) as internal standard. Milk is extracted with acetonitrile, centrifuged, evaporated, reconstituted in water, and passed through a C18 solid-phase extraction (SPE) column. The SPE column is eluted with 60% methanol, and then the eluate is evaporated and derivatized with Sylon BFT ¿N,O-bis(trimethylsilyl)trifluoroacetamide [BSTFA]-trimethylchlorosilane [TMCS], 99 + 1¿. After derivatization, toluene is added directly to the sample, followed by water, to quench the derivatization process. After centrifugation, the organic layer is carefully removed. Analytes are determined by GC with electron capture detection (ECD). Milk was fortified with fenicols (the collective name for CAP, FF, and TAP) at 5, 10, 20, 40 and 80 ng/mL (target level = 10 ng/mL). Overall recoveries were 92, 100, and 104% for CAP, FF, and TAP, respectively. Overall interassay (between-day) variabilities were 6.1, 6.7, and 6.0% for CAP, FF, and TAP, respectively. Raw milk samples containing incurred residues of FF were also analyzed.