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Abstract
A new reverse phase ultra-performance liquid chromatography (RP-UPLC) method was developed for the quantification of bisoprolol hemifumarate (BIS) and hydrochlorothiazide (HCT) in tablets using experimental design and optimization. In the development of method, a 33 full factorial design was used to identify the effects of factors (column temperature, flow rate, and 0.1 M H3PO4 % in mobile phase) and their interactions on the chromatographic response and to determine optimal chromatographic conditions providing a good peak elution and shortest runtime, respectively. For the optimal RP-UPLC conditions, 58.2°C (column temperature), 0.37 mL/min (flow rate) and 23.6% (0.1 M H3PO4) were obtained from the results of 33 full factorial design. The separation of BIS (retention time: 0.323 min) and HCT (retention time: 0.409 min) was performed within a short runtime of 0.6 min. Calibration curves for BIS and HCT in the range 8.0–40.0 µg/mL and 2.0–26.0 µg/mL were computed by the detection at 225 nm. The validity of the method was achieved by analyzing the test samples, and using inter-days and intra-day studies. Under optimal conditions, new RP-UPLC method was successfully applied for the analysis of BIS and HCT in commercial tablets.