Liquid-chromatographic detection of thiazide diuretics in urine

We describe a liquid-chromatographic procedure for detection in urine of all thiazide drugs currently used clinically. Urine is treated initially with NaBH4 to convert any chlorothiazide present to hydrochlorothiazide. The urine is acidified with NaH2PO4 (1.0 mol/L, pH 5), and thiazides are extracted with ethyl acetate. Interfering substances are then removed in two washes with 0.1 mol/L Na2HPO4 at pH 8. The ethyl acetate is evaporated and the residue redissolved in mobile phase. Thiazides are assayed by reversed-phase chromatography, with detection by ultraviolet absorption. Analytical recovery of thiazides ranged from 53 to 93%. Urines from 48 patients were so studied, and the results were compared with results by the currently used spectrophotometric method. The two methods agreed for 56% of samples. Evaluation of the discrepancies by review of patients' histories clearly showed liquid chromatography to have correctly identified seven of eight positive urines that the spectrophotometric method failed to detect. Ultraviolet scanning incorrectly identified as positive two samples, whereas liquid chromatography did not falsely identify any urines as positive. Our method was more sensitive and more specific than the spectrophotometric method.