title: Development of an Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of Amoxicillin in Broth Medium and its Application to an In Vitro Pharmacokinetic and Pharmacodynamic Model.
abstract: A simple, rapid and highly sensitive liquid chromatographic-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the quantification of amoxicillin in broth-a liquid bacterial culture medium. After appropriate dilution with ultrapure water, broth samples containing amoxicillin and an internal standard (IS) were extracted by acetonitrile and dichloromethane. The extract was injected into the system. The analyte and the IS were separated by a prepacked Atlantis C18 column using acetonitrile-0.1% formic acid as a mobile phase and detected by selected reaction monitoring in electrospray ionization positive ion mode. The calibration curve of amoxicillin was linear over the concentration range of 0.05-20.00 µg/mL. The mean recovery of amoxicillin from broth was 71.7%, and the intra- and interday precision and accuracies of the assay were within 10%. Amoxicillin was stable in broth for 12 h at room temperature (24°C), for 6.5 months at -80°C and for 24 h after preparation in an autosampler at room temperature. It has been successfully applied to an in vitro pharmacokinetic (PK) and pharmacodynamic (PD) model in which the broth is used for bacterial growth. The method provides high-throughput biological analysis to facilitate the in vitro PK and PD model of amoxicillin.
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Shujing
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China fanyang9@fudan.edu.cn.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Beining
authors.initials: B
authors.lastname: Guo
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Yuancheng
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Wu
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Liang
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Yaoguo
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Institute of Antibiotics, Huashan Hospital, Fudan University and Key Laboratory of Clinical Pharmacology of Antibiotics, Ministry of Health, 12, Wulumuqi Zhong Road, Shanghai 200040, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000658:Amoxicillin / Q000008:administration & dosage / Q000493:pharmacokinetics*; D000900:Anti-Bacterial Agents / Q000008:administration & dosage / Q000493:pharmacokinetics*; D001419:Bacteria / Q000187:drug effects / Q000254:growth & development; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D006801:Humans; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000900:Anti-Bacterial Agents; D003470:Culture Media; D000658:Amoxicillin