Analysis of Hydrocholorothiazide by Using Ultra Performance Liquid Chromatography

Authors : Anupama Maurya, Urmila Nishad, Meraj Al, Manoj Kumar Yadav, Vijay Kumar Yadav

Volume/Issue : Volume 5 - 2020, Issue 3 - March

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


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