Antibiotic Absorption In Zebrafish Using Liquid Chromatography-tandam Mass Spectrometry

Drug toxicity is a master reason for drug withdrawal from the market, safety and efficacy limits R&D and clinical use for compounds. Therefore, accurate toxicity assessment can increase the feasibility of a drug reaching consumers at the earty satge of R&D. Zebrafish (Danio rerio) is gaining attention as a primary drug toxicity screening model. Due to drug absorption variances in zebrafish, the study aim to establish a rapid, sensitive and reliable LC-MS/MS method using an MRM mode, and quantification of antibiotics in zebrafish for drug absorption evaluation in toxicity testing. This study mainly refers to several aspects as following:The first part is eatablish methodologies for drugs in zebrafish by liquid chromatography-tandem Mass spectrometry. The method was scaned by MRM modes, and method validation was completed according to guidelines for validation of bioanalytical methods, and establish cephalosporin MS database, sMRM-IDA-EPI method is applied to analyze chemisty in zebrafish toxicity testThe second part is mainly about drug absorption and toxicity in zebrafish by LC-MS/MS, including (1) dose of drugs absorption in different zebrafish period, and zebrafish toxic effect and phenotype at different dose;(2) quantitative determination of cephalosporin and aminoglycoside in zebrafish by LC-MS/MS;(3) according to value of Ka, forecasting the drugs absorption and validation by ADMET Predictor.Our results shown that a rapid, sensitive and reliable LC-MS/MS method using MRM for compounds of different polarities was established for quantification of compound absorption in zebrafish. Zebrafish embryos absorbed compounds uniquely depending on the development period, and there are close relationship between toxicity and absorption. Our novel approach may help interpreting zebrafish toxicity data. Quantification of target compounds can better estimate toxicity in zebrafish toxicity tests.