A High-Performance Liquid Chromatography/Negative Ion Electrospray Tandem Mass Spectrometry Method for the Determination of Hydrochlorothiazide in Human Plasma: Application to a Comparative Bioavailability Study

Abstract

A sensitive, selective and rapid liquid chromatography/tandem mass spectrometric (LC–MS/MS) method was developed and validated for the determination of hydrochlorothiazide (HCTZ) in human plasma. The plasma samples were prepared by solid-phase extraction using Oasis HLB 30 mg/1CC cartridges. Chromatographic separation was accomplished on a Thermo Hypurity Advance (50 mm × 4.6 mm i.d., 5 μm) column. The mobile phase consisted of HPLC-grade acetonitrile: 2 mM ammonium acetate (90: 10 v/v) at a flow rate of 0.5 mL min⁻¹. Detection of hydrochlorothiazide and the internal standard (IS) zidovudine was achieved by electrospray ionization (ESI) MS/MS in the negative ion mode. The total chromatographic run time was 2.5 min. The linear range of the method was from 2.036–203.621 ng mL⁻¹. The mass transition ion pair has been followed as m/z 296.10/205.00 for HCTZ and m/z 266.10/223.10 for zidovudine. The mean overall recovery of HCTZ was 66.40% with a precision of 2.44%. The mean recovery of the internal standard (zidovudine) was 63.62% with a precision ranging from 2.06% to 5.40%. The method was successfully applied to the evaluation of pharmacokinetics of hydrochlorothiazide after single oral dose of 25 mg hydrochlorothiazide to healthy volunteers.

Keywords

hydrochlorothiazide zidovudine solid-phase extraction LC-MS-MS

Get full access to this article

View all access options for this article.

References

Lant

, “Diuretics. Clinical pharmacology and therapeutic use (Part II)”, Drugs 9, 162 (1985). doi: 10.2165/00003495-198529020-00003

Trenkwalder

, “Combination therapy with AT (1)-receptor blockers”, J. Hum Hypertens. 16, S17 (2002). doi: 10.1038/sj.jhh.1001435

Oparil

Aurup

Snavely

Goldberg

, “Efficacy and safety of losartan/hydrochlorothiazide in patients with severe hypertension”, Am. J. Cardiol. 87, 721 (2001). doi: 10.1016/S0002-9149(00)01490-9

Gradman

A.H.

Brady

W.E.

Gazdick

L.P.

Lyle

Zeldin

R.K.

, “A multicenter, randomized, double-blind, placebo-controlled, 8-week trial of the efficacy and tolerability of once-daily losartan 100 mg/hydrochlorothiazide 25 mg and losartan 50 mg/hydrochlorothiazide 12.5 mg in the treatment of moderate-to-severe essential hypertension”, Clin. Ther. 24, 1049 (2002). doi: 10.1016/S0149-2918(02)80018-2

Huang

Yang

Shao

Zheng

Duan

, “Simultaneous determination of captopril and hydrochlorothiazide in human plasma by reverse-phase HPLC from linear gradient elution”, J. Pharmaceut. Biomed. Anal. 41, 644 (2006). doi: 10.1016/j.jpba.2005.12.007

Takubo

Okada

Ishii

Hara

K.-I.

Ishii

, “Sensitive and selective liquid chromatography–electrospray ionization tandem mass spectrometry analysis of hydrochlorothiazide in rat plasma”, J. Chromatogr. B 806, 199 (2004). doi: 10.1016/j.jchromb.2004.03.060

Wang

Jiang

Tang

Wang

Zhao

, “A liquid chromatography/tandem-mass spectrometry method for the simultaneous quantification of valsartan and hydrochlorothiazide in human plasma”, J. Chromatogr. B 852, 436 (2007). doi: 10.1016/j.jchromb.2007.02.014

Liu

Gao

Zhang

Guo

, “Determination of hydrochlorothiazide in human plasma by liquid chromatography/tandem mass spectrometry”, J. Pharmaceut. Biomed. Anal. 44, 1187 (2007). doi: 10.1016/j.jpba.2007.04.020

Song

Hang

Zhao

Wang

, “Simultaneous determination of amiloride and hydrochlorothiazide in human plasma by liquid chromatography/tandem-mass spectrometry with positive/negative ion-switching electrospray ionization”, Rapid Commun. Mass Spectrom. 21, 3427 (2007). doi: 10.1002/rcm.3235

10.

Guidance for Industry, Bioanalytical Method Validation. US Department of Health and Human Services, Food and Drug Administration, Centre for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM), May (2001).

11.

FDA Guidance for Industry: Bioavailability and Bioequivalence Studies for Orally Administered Drug-Products-General Considerations. US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), March (2003).