Validated LC-MS/MS method for simultaneous analysis of 21 cephalosporins in zebrafish for a drug toxicity study

Zebrafish model was used to perform this drug toxicity study. In this study, we monitored the absorption of drugs in zebrafish. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed in this experiment. The LC-MS/MS method was used for the simultaneous quantification of 21 cephalosporins in zebrafish. To prepare samples, rapid protein precipitation was performed with 1% formic acid in methanol. All the analytes were separated on a C₁₈ column. These analytes were analyzed by performing multiple-reaction monitoring (MRM) scans in positive electrospray ionization mode. The ranges for limits of detection (LOD) and quantification (LOQ) were as follows: 0.01-10 ng/mL (S/N ≥ 3) and 0.05-25 ng/mL. Intra-and inter-day accuracy of all 21 cephalosporins were within the range of 80%-120%. Intra-and inter-day precision were within the range of 0.4%-11.2%. All the 21 analytes were analyzed within 10 min. In drug toxicity studies, the present method can be used to determine the internal concentrations of cephalosporins in zebrafish following the administration of drugs.