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Development and Validation of a RP-HPLC Method for the Simultaneous Determination of Rifampicin and a Flavonoid Glycoside - A Novel Bioavailability Enhancer of Rifampicin
Abstract
Purpose: To develop and validate a sensitive HPLC method for the separation and simultaneous estimation of two ingredients in a composition comprising of rifampicin and a flavonoid glycoside (an
enhancer of oral bioavailability of rifampicin). Methods: Reverse phase (RP) chromatographic separation and estimation was achieved using a Shimadzu HPLC system. RP-18 column was used at the following optimised conditions: mobile phase, acetonitrile:phosphate buffer, 50 mM, pH 5.0 in a ratio of 60:40 v/v; oven temperature, 40 0C; flow rate, 0.8 ml min-1; detection wavelength, 340 nm; and total run time, 15 min. Results: The developed method was validated in terms of linearity, range, accuracy, precision, limit of detection, limit of quantification, robustness and specificity. Good linearity was observed (r2 > 0.999) over the study range of both ingredients. The precision values for rifampicin and the flavonoid glycoside were in the range 1.08-2.77 and 1.14-2.98 %, respectively, while the limit of quantification was 0.10 and 0.05μg mL-1 respectively. The method was found to be robust and specific for both ingredients. Conclusion: The developed method has a potential application in preclinical and clinical studies.
Keywords: Flavonoid glycoside; RP-HPLC; Rifampicin; Stability studies; Validation
