Immunoaffinity Column Cleanup with LC/MS/MS for the Determination of Chloramphenicol in Honey and Prawns: Single-Laboratory Validation

Abstract

Abstract A single-laboratory validation was conducted to establish the effectiveness of an immunoaffinity column (IAC) cleanup procedure followed by LC/MS/MS for the determination of chloramphenicol (CAP) in honey and prawns. Honey is dissolved in buffer solution and centrifuged, and an aliquot applied to an IAC. For prawns, a portion of the homogenized sample is shaken with buffer and centrifuged, and an aliquot similarly applied to an IAC. For both matrix extracts, CAP is removed from the IAC with neat methanol, then directly analyzed by electrospray LC/MS/MS in the negative ionization mode using m/z 321 as a precursor ion and m/z 257 and 152 as qualifier and quantifier ions, respectively. Test portions of blank honey and prawns were fortified with CAP to give levels of 0.3, 1.0, and 5.0 μg/kg. Recoveries of CAP on 3 consecutive days ranged from 83–103% for honey and 84–108% for prawns. Based on results for fortified blank matrixes (triplicate at three levels), the RSD for repeatability (RSDr) averaged 8.4% for honey and 4.8% for prawns. The method LOD was 0.05 for prawns and 0.16 μg/kg for honey, both well below the minimum required method performance limit for CAP. The accuracy of the method was demonstrated by participation in proficiency testing, where satisfactory Z-scores were obtained for CAP in incurred samples of both honey and prawns. The method was shown to be applicable to a wide range of other matrixes, including milk, egg, royal jelly, meat, and seafood products.