HPLC–DAD Analysis of Hydrochlorothiazide and Irbesartan in Hypertensive Patients on Fixed-Dose Combination Therapy
2011
Hydrochlorothiazide (HCTZ) and the angiotensin II type 1 receptor antagonist (ARB) irbesartan (IRBE) are well-known antihypertensive drugs, frequently administered as a low-dose combination in a single pill. In this work, a simple, sensitive, and selective high-performance liquid chromatographic (HPLC) method with diode-array detection was developed for simultaneous determination of HCTZ and IRBE levels in the plasma of hypertensive patients given a fixed combination of 12.5 mg HCTZ and 300 mg IRBE. Compounds were extracted from acidified plasma samples with 3 mL ethyl acetate, and eluted at 6 and 19 min from a C4 column by elution with an acetonitrile–phosphate buffer (pH 3.6) mobile-phase gradient at a flow rate of 1 mL min−1. The assay was linear over the ranges 2.5–500 and 20–4,000 ng mL−1 for HCTZ and IRBE, respectively. Overall intra-assay and inter-assay variation were within acceptance limits. Limits of quantification were 2.5 and 20 ng mL−1 for HCTZ and IRBE, respectively. Plasma samples remained stable for 12 h at room temperature, through three thaw–freeze cycles, and for 2 and 7 months at −20 °C. In hypertensive patients, residual concentrations were 22.3 ± 6.0 and 241.8 ± 39.0 ng mL−1 for HCTZ and IRBE, respectively. There was no interference from other co-administered drugs. Despite the different physicochemical properties of HCTZ and IRBE, our method enables accurate measurement of both drugs for assessment of compliance by patients treated by fixed-dose combination therapy with HCTZ–IRBE.
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