Simple determination of hydrochlorothiazide in human plasma and urine by high performance liquid chromatography

The diuretic drug hydrochlorothiazide (HCT) is used mainly for treatment of mild to moderate hypertension and is usually administered with other drugs. An assay for the determination of HCT in human plasma and urine by high performance liquid chromatography (HPLC) has been developed. Samples were purified by solvent extraction and analysed by reversed phase HPLC with ultraviolet detection, using hydroflumethiazide as the internal standard; plasma was eluted using gradient elution and urine was analysed isocratically. The method is simple to perform, is sensitive (detection limit 0.01 micrograms/mL in plasma and 0.2 micrograms/mL for urine); it showed good reproducibility (3-8%). A great number of drugs did not interfere with the assay and the method was used for pharmacokinetic studies in healthy subjects, but samples from patients can also be analysed with high selectivity.