A simple, specific, sensitive, precise, and accurate high performance liquid chromatography procedure was developed and validated for the analysis of hydrochlorothiazide in human plasma. After addition of hydroflumethiazide as internal standard, the analytes were isolated from human plasma by double liquid-liquid extraction and separated on a reversed phase column with acetonitrile/water 0 : 80, v/v as the eluent. Peaks were monitored at 271 nm. Peak heights were measured and hydrochlorothiazide was quantified using calibration curves. The method was found to be linear in the 5 to 80 ng/mL concentration range (r<0.999). The limit of quantitation was found to be 5 ng/mL for 1 mL plasma samples. The inter-assay precision, expressed as the coefficients of variation (CV%), ranged from 2.7 to 9.4% and the assay inaccuracy was within 2.2%. The method was applied to estimate the pharmacokinetics of hydrochlorothiazide after administration of a single oral dose containing 20 mg enalapril and 12.5 mg hydrochlorothiazide to 24 healthy volunteers.
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Niopas et al. (2002) studied this question.