title: Analysis of benzodiazepines and their metabolites using DBS cards and LC-MS/MS.
abstract: Dried Blood Spot (DBS) has been used a blood extraction method for inherited metabolic disorder screening since 1960s. With introduction of LC-MS/MS, not only DBS could be used to analysis drugs in small blood volume, but in various fields, such as toxicology, drug therapeutic monitoring, drug diagnostic screening, and illicit drugs. In toxicology field, many drugs (e.g. benzodiazepines, acetaminophen, small molecule drugs) have been tested with DBS. Compared with earlier blood extraction methods (SPE and LLE), DBS has lots of advantages; lower blood volume (less than 50μL), shorter analysis time caused by a more concise analysis procedure and lower cost. We optimized the DBS procedure and LC-MS/MS conditions for 18 benzodiazepines, seven benzodiazepine metabolites, and one z-drug (zolpidem) analysis in blood. 30μL of whole blood was spotted on FTA DMPK card C and dried for 2h in a desiccator. A 6-mm disk was punched and vortexed for 1min in a centrifuge tube with 300μL methanol/acetonitrile mixture (1:1, v/v). After evaporation, redissolved in 100μL mobile phase of LC-MS/MS and 5μL was injected. In the analysis for 26 target compounds in blood, all of the method validation parameters - LLOD, LLOQ, accuracy (intra- and inter-assay), and precision (intra- and inter-assay) - were satisfied with method validation criteria, within 15%. The results of matrix effect, recovery, and process efficiency were good. We developed a fast and reliable sample preparation method using DBS for 26 benzodiazepines, benzodiazepine metabolites, and z-drug (zolpidem).
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Heesang
authors.initials: H
authors.lastname: Lee
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Yujin
authors.initials: Y
authors.lastname: Park
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Jiyeong
authors.initials: J
authors.lastname: Jo
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Sangwhan
authors.initials: S
authors.lastname: In
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Yonghoon
authors.initials: Y
authors.lastname: Park
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea.
authors.forename: Eunmi
authors.initials: E
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Kyungsung University, 309 Suyeongro, Namgu, Busan, Republic of Korea.
authors.forename: Jaesung
authors.initials: J
authors.lastname: Pyo
authors.affiliation: Busan Institute, National Forensic Service, 50 Geumhoro, Mulgeumeup, Yangsan, Republic of Korea. Electronic address: csk91@korea.kr.
authors.forename: Sanggil
authors.initials: S
authors.lastname: Choe
mesh_terms: D001569:Benzodiazepines / Q000097:blood*; D002853:Chromatography, Liquid; D059788:Dried Blood Spot Testing; D053593:Forensic Toxicology / Q000379:methods; D006801:Humans; D006993:Hypnotics and Sedatives / Q000097:blood; D013058:Mass Spectrometry; D011725:Pyridines / Q000097:blood; D000077334:Zolpidem
chemical_list: D006993:Hypnotics and Sedatives; D011725:Pyridines; D001569:Benzodiazepines; D000077334:Zolpidem
keywords: Benzodiazepines; Dried Blood Spot; LC–MS/MS; Method validation