title: Development and validation of a hydrophilic interaction liquid chromatography-tandem mass spectrometry method for the simultaneous determination of five first-line antituberculosis drugs in plasma.
abstract: A new, sensitive and fast method for the simultaneous determination of pyrazinamide, isoniazid, streptomycin, ethambutol, and rifampicin in human plasma was developed and validated. The method required only 100 μL of plasma and one step for sample preparation by protein precipitation. The drugs were separated by using a hydrophilic interaction liquid chromatography (HILIC) column. The mobile phase was methanol and water (0.1% formic acid and 5 mM ammonium acetate, pH 3.0 ± 0.1) in a ratio of 65:35 (v/v), which was eluted at an isocratic flow rate of 0.5 mL/min. Tandem mass spectrometry was performed with a triple-quadrupole tandem mass spectrometer. By use of the HILIC column, the detection was free of ion-pair reagents in the mobile phase, with no significant matrix effects. The total run time was less than 2 min for each sample. The method was validated by evaluating its selectivity, sensitivity, linearity, accuracy, and precision according to US Food and Drug Administration guidelines. The lower limit of quantification was 4.0 ng/mL for pyrazinamide, isoniazid, and rifampicin, 0.5 ng/mL for ethambutol, and 10.0 ng/mL for streptomycin. The intraday precision and interday precision were less than 9%, with the accuracy ranging between -9.3 and 7.3%. The method was successfully applied to therapeutic drug monitoring of 33 patients with tuberculosis after administration of standard antituberculosis drugs. The method has been proved to meet the high-throughput requirements in therapeutic drug monitoring.
authors.affiliation: Department of Health Inspection and Quarantine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Zhou
authors.forename: Xianbo
authors.initials: X
authors.lastname: Wu
authors.forename: Qinzhi
authors.initials: Q
authors.lastname: Wei
authors.forename: Yungang
authors.initials: Y
authors.lastname: Liu
authors.forename: Peng
authors.initials: P
authors.lastname: Liu
authors.forename: Ande
authors.initials: A
authors.lastname: Ma
authors.forename: Fei
authors.initials: F
authors.lastname: Zou
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000995:Antitubercular Agents / Q000097:blood*; D002648:Child; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D016903:Drug Monitoring; D004977:Ethambutol / Q000097:blood; D005260:Female; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007538:Isoniazid / Q000097:blood; D008297:Male; D008875:Middle Aged; D011718:Pyrazinamide / Q000097:blood; D012293:Rifampin / Q000097:blood; D013307:Streptomycin / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014376:Tuberculosis / Q000097:blood* / Q000188:drug therapy; D055815:Young Adult
chemical_list: D000995:Antitubercular Agents; D011718:Pyrazinamide; D004977:Ethambutol; D007538:Isoniazid; D012293:Rifampin; D013307:Streptomycin