Development and
Validation of a RP-HPLC Method for the Simultaneous
Determination of Rifampicin and a Flavonoid Glycoside -
A Novel Bioavailability Enhancer of Rifampicin
Bhusari S Sachin1, Vandhna Bhat1,
Meenakshi Koul1, Subhash C Sharma1,
Manoj K Tikoo1, Ashok K Tikoo1,
Naresh K Satti2, Krishan A Suri2
and Rakesh K Johri1*
Divisions of 1Pharmacology
and 2Natural Products Chemistry, Indian
Institute of Integrative Medicine, Jammu 180001, J & K,
India.
*Corresponding author:
E-mail:
rkjohri@iiim.res.in
Tel: +91-9419132014
Received: 30 April 2009
Revised
accepted: 15 September 2009
Tropical
Journal of Pharmaceutical Research, December 2009; 8(6):
531-537
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
