title: Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.
abstract: As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 μg/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (ρ 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment.
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|Department of Laboratory Medicine, Seoul National University Hospital, Seoul, Korea.
authors.forename: Kyunghoon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Jun
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|Department of Laboratory Medicine, Sheikh Khalifa Specialty Hospital, Ras Al Khaimah, United Arab Emirates.
authors.forename: Minje
authors.initials: M
authors.lastname: Han
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|Department of Laboratory Medicine, Seoul National University Hospital, Seoul, Korea.
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Song
authors.affiliation: Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
authors.forename: Jong Sun
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
authors.forename: Kyoung Un
authors.initials: KU
authors.lastname: Park
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea. songjhcp@snu.ac.kr.
authors.forename: Junghan
authors.initials: J
authors.lastname: Song
mesh_terms: D000995:Antitubercular Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D059788:Dried Blood Spot Testing; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
chemical_list: D000995:Antitubercular Agents
keywords: Anti-tuberculosis drug; Dried blood spot; Multiplex analysis; Tandem mass spectrometry; Therapeutic drug monitoring