Ann Lab Med.  2016 Sep;36(5):489-493. 10.3343/alm.2016.36.5.489.

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

Affiliations
  • 1Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea. songjhcp@snu.ac.kr
  • 2Department of Laboratory Medicine, Seoul National University Hospital, Seoul, Korea.
  • 3Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
  • 4Department of Laboratory Medicine, Sheikh Khalifa Specialty Hospital, Ras Al Khaimah, United Arab Emirates.
  • 5Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.

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.

Keyword

Anti-tuberculosis drug; Tandem mass spectrometry; Dried blood spot; Multiplex analysis; Therapeutic drug monitoring

MeSH Terms

Antitubercular Agents/*blood
Chromatography, High Pressure Liquid
*Dried Blood Spot Testing
Humans
Limit of Detection
Reproducibility of Results
*Tandem Mass Spectrometry
Antitubercular Agents

Figure

  • Fig. 1 Passing-Bablok regression with regression equations, Pearson's rho, significance levels and Bland-Altman plots between measurements in DBS and plasma for second-line anti-TB drugs: kanamycin, cycloserine, moxifloxacin, levofloxacin, prothionamide, PAS, linezolid, and clarithromycin.Abbreviations: DBS, dried blood spot; PAS, para-aminosalicylic acid.


Cited by  1 articles

Recommendations for the Use of Liquid Chromatography-Mass Spectrometry in the Clinical Laboratory: Part I. Implementation and Management
Kyunghoon Lee, Soo Young Moon, Serim Kim, Hyun-Jung Choi, Sang-Guk Lee, Hyung-Doo Park, Soo-Youn Lee, Sang Hoon Song,
Lab Med Online. 2020;10(1):1-9.    doi: 10.3343/lmo.2020.10.1.1.


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