title: Development, validation and clinical application of an online-SPE-LC-HRMS/MS for simultaneous quantification of phenobarbital, phenytoin, carbamazepine, and its active metabolite carbamazepine 10,11-epoxide.
abstract: A simple and efficient bioanalytical method for simultaneous determination of phenobarbital (PB), phenytoin (PHT), carbamazepine (CBZ), and its active metabolite carbamazepine 10,11-epoxide (CBZE) in human plasma using online solid phase extraction (SPE)-liquid chromatography (LC) coupled with high resolution mass spectrum (HRMS) under targeted MS/MS (t-MS(2)) analysis mode has been developed. The procedure integrated an automated sample clean-up of human plasma by Oasis®HLB SPE cartridge, a separation by ZORBAX SB-C18 analysis column, and a quantification by Q-Exactive Hybrid Quadrupole-Orbitrap. The total running time was 13min. The lower limit of quantification (LLOQ) of PB, PHT, CBZ, and CBZE were 0.008, 0.008, 0.0016 and 0.0016μgmL(-1) respectively and the linearities were in the range of 0.008-2.500, 0.008-2.500, 0.0016-0.500 and 0.0016-0.500μgmL(-1) respectively. The mean recovery was between 91.82% and 108.27% and the matrix effect was between 93.29% and 102.09%. The relative standard deviations of interday and intraday were less than 6.41%. The method has been successfully applied in therapeutic drug monitoring (TDM) of four Chinese epilepsy patients. This fully automated, simple, sensitive and reliable online-SPE-LC-HRMS/MS method serves well for TDM of PB, PHT, CBZ and CBZE at clinics for either single or combination treatment.
authors.affiliation: Center of Basic Molecular Science (CBMS), Department of Chemistry, Tsinghua University, Beijing 100084, PR China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Qu
authors.affiliation: Core Facilities of Madern Pharmaceuticals, Core Facilities Center, Capital Medical University, Beijing 100069, PR China. Electronic address: 714031230@qq.com.
authors.forename: Yuanjie
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Center of Basic Molecular Science (CBMS), Department of Chemistry, Tsinghua University, Beijing 100084, PR China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wang
authors.affiliation: Beijing Institute of Functional Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing 100053, PR China. Electronic address: makaipeter@sina.com.
authors.forename: Kai
authors.initials: K
authors.lastname: Ma
authors.affiliation: Center of Basic Molecular Science (CBMS), Department of Chemistry, Tsinghua University, Beijing 100084, PR China. Electronic address: yinzheng@mail.tsinghua.edu.cn.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Yin
mesh_terms: D000328:Adult; D000927:Anticonvulsants / Q000097:blood*; D002220:Carbamazepine / Q000031:analogs & derivatives* / Q000097:blood* / Q000493:pharmacokinetics / Q000627:therapeutic use; D002851:Chromatography, High Pressure Liquid; D016903:Drug Monitoring; D004827:Epilepsy / Q000097:blood / Q000188:drug therapy; D005260:Female; D006801:Humans; D008297:Male; D010634:Phenobarbital / Q000097:blood* / Q000493:pharmacokinetics / Q000627:therapeutic use; D010672:Phenytoin / Q000097:blood* / Q000493:pharmacokinetics / Q000627:therapeutic use; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000927:Anticonvulsants; D002220:Carbamazepine; D010672:Phenytoin; C012227:carbamazepine epoxide; D010634:Phenobarbital
keywords: Antiepileptic drugs; Online-SPE-LC-HRMS/MS; Therapeutic drug monitoring