Confirmation of Chloramphenicol Residues in Egg by Gas Chromatography/High-Resolution Mass Spectrometry and Comparison of Quantitation with Gas Chromatography–Electron Capture Detection

Abstract

Determination of chloramphenicol (CAP) residues in egg by gas chromatography/high-resolution mass spectrometry (GC/HRMS) with negative chemical ionization and gas chromatography with electron capture detection (GC-ECD) is described. A cleanup based on acetonitrile extraction followed by solid-phase extraction with silica gel and gel filtration columns was developed for extraction of CAP residues from whole egg. For quantitation, the internal standards used were the meta isomer of CAP (m-CAP) for GC-ECD and both m-CAP and deuterium-labeled CAP (D5-CAP) for GC/HRMS. For GC/HRMS, evaluation was performed by selecting characteristic ions at m/z 466 for CAP and m-CAP and at m/z 471 for D5-CAP. Both methods were validated with egg samples fortified at 0.4-2.0 micrograms/kg and lyophilized egg samples from animals treated at the 1 microgram/kg concentration level. The coefficient of variation was below 10%. Limits of detection and quantitation of both methods were about 0.3 and 0.5 microgram/kg, respectively. For confirmation of CAP residues, the relative ion abundance (m/z 466 and 468) was calculated. Further confidence was obtained by comparison of accurate monoisotopic masses from a spectrum library.