Development and validated RP-UPLC method for simultaneous estimation of ciprofloxacin HCl, doxycycline and phenazopyridine HCl in bulk and tablet dosage form.

A Reversed-Phase Ultra Performance liquid Chromatographic (RP-UPLC) method was developed for the simultaneous determination of Ciprofloxacin HCL, Doxycycline Hyclate  and Phenazopyridine HCL in tablet dosage form. The analysis was carried out using Acquity UPLC, BEH C–18, 50 X 2.1, 1.7µ column. Mobile phase, containing 0.05 M Ammonium Acetate Buffer: Methanol (50:50) pH adjusted to 4 with Ortho-phosphoric acid was pumped at a flow rate of 1mL/min with UV-detection at 278,350,378 nm Respectively. Retention time was 0.90 ± 0.01 min, 1.60±0.02 min & 4.17±0.01 min. for Ciprofloxacin HCL, Doxycycline and Phenazopyridine HCL respectively. The method was validated for linearity, accuracy, precision, and specificity. The method showed good linearity in the range 20, 50, 80,100, 120, 150, 200 ppm. The % R.S.D for precision and accuracy of the method was found to be less than 2%. The method was validated as per the ICH guidelines. The method was successfully applied for routine analysis of Ciprofloxacin HCL, Doxycycline and Phenazopyridine HCL in combined tablet dosage form.