Liquid chromatography–mass spectrometry method for the determination of thiamphenicol in rabbit tears

Abstract

A simple and sensitive liquid chromatography–mass spectrometry (LC–MS) method was described for the determination of thiamphenicol in rabbit tears. Chromatographic separation of the analyte was achieved on a C18 column using a mobile phase of acetonitrile and 10 mmol/l ammonium acetate solution (60:40, v/v). Selected ion monitoring (SIM) in negative mode was used for analyte quantification at m/ z 354.4 for thiamphenicol and at m/ z 137.1 for salylic acid. The run time was less than 6 min. Linearity over the concentration range of 0.032–32.0 ng/ml for thiamphenicol was obtained and the lower limit of quantification was 0.032 ng/ml. For each level of QC samples, inter- and intra-day precisions (R.S.D.) were ≤5.2% and 8.3%, respectively, and the accuracy (RE) was ±2.8%. The present LC–MS method was successfully applied to the pharmacokinetic studies of thiamphenicol in situ forming gel in rabbit tears.