Abstract
A HPLC method with a very sensitive electrochemical detection has been developed for determining the diuretic agent hydrochlorothiazide (HCT) in serum of volunteers to whom a single dose of the fixed combination of 12.5 mg HCT and 25 mg triamterene was administered. In the present method samples (0.2 ml serum, pH 7) were purified by extraction of HCT into 5 ml tert.-butyl-methyl ether. The separate organic phase was spiked with p-aminobenzoic acid (PABA) standard. The separation was performed on a reversed-phase C 18 column using phosphate buffer-acetonitrile (90:10). A coulometric cell was used to measure HCT and an ultraviolet detector for PABA. The limit of quantitation for serum samples of only 200 μl was 5 ng/ml (i.e. 60 pg HCT/injection) with a good reproducibility (1–8%). Short retention times were found: 1.2 min for PABA and 5.8 min for HCT.
Published Version
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