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Anupama Maurya*, Urmila Nishad, Deeksha Chaturvedi, Manoj Kumar Yadav and Vijay Kumar Yadav


A new, simple, accurate and stability-indicating UPLC (Ultra Performance Liquid Chromatography) method was developed and simultaneously validated for determination of Diuretic Drug of thiazide class of Hydrochlorothiazide and Chlorothiazide in urine sample. It involved 150 mm × 2.1 mm, Acclaim column. The separation was obtained on the simple gradient method. The mobile phase was composed of 0.2% TFA in water and 0.2% TFA in acetonitrile. The flow rate of the mobile phase for gradual elution was set to 0.5 ml / min-1 and the column temperature was maintained at 40˚C. The detector of wavelength was 271 nm for Hydrochlorothiazide and 271nm for Chlorothiazide. The retention times of Hydrochlorothiazide and Chlorothiazide are 2.9 and 3.4 minutes; respectively. The total run time was 6.0 minutes within which the two compounds were and separated. The described method was valid with respect to system linearity, precision, and accuracy. The accuracy of the assay method was evaluated by six independent assays Hydrochlorothiazide and Chlorothiazide. The described method was linear over the range, 0.625 to 10 μg mL-1 for Chlorothiazide, and 0 .625 to 10 μg·mL-1 for Hydrochlorothiazide. The detection limit for HCTZ 0.4602580 and for CTZ 0.492828, LOQ for HCTZ 1.534194 CTZ 1.642759 the method was validated as linearity, accuracy, and reproducibility. Recovery was in the range of 93 - 108%. The method is rapid and suitable for the analysis of hydrochlorothiazide and Chlorothiazide in urine samples.

Keywords: Chlorothiazide, Hydrochlorothiazide, Method Development etc.

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