Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

• Published in: Annals of laboratory medicine Vol. 36; no. 5; pp. 489 - 493
• Main Authors: Lee, Kyunghoon, Jun, Sun-Hee, Han, Minje, Song, Sang Hoon, Park, Jong Sun, Lee, Jae Ho, Park, Kyoung Un, Song, Junghan
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society for Laboratory Medicine 01.09.2016; 대한진단검사의학회
• Subjects: Antitubercular Agents - blood; Brief Communication; Chromatography, High Pressure Liquid; Dried Blood Spot Testing; Humans; Limit of Detection; Reproducibility of Results; Tandem Mass Spectrometry; 병리학; Therapeutic drug monitoring; Anti-tuberculosis drug; Tandem mass spectrometry; Dried blood spot; Multiplex analysis
• ISSN: 2234-3806; 2234-3814; 2234-3814
• DOI: 10.3343/alm.2016.36.5.489

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.