Background: High performance liquid chromatography is an integral analytical
tool in assessing drug product stability. HPLC methods should be able to separate,
detect, and quantify the various drug-related degradants that can form on storage
or manufacturing, plus detect any drug-related impurities that may be introduced
Objective: A simple, economic, selective, precise, and stability-indicating HPLC
method has been developed and validated for analysis of Rifampicin (RIFA) and
Piperine (PIPE) in bulk drug and in the formulation.
Method: Reversed-phase chromatography was performed on a C18 column with
Buffer (Potassium Dihydrogen Orthophosphate) pH 6.5 and Acetonitrile, 30:70), (%,
v/v), as mobile phase at a flow rate of 1 mL min-1.
Result: The detection was performed at 341 nm and sharp peaks were obtained for
RIFA and PIPE at retention time of 3.3 ± 0.01 min and 5.9 ± 0.01 min, respectively.
The detection limits were found to be 2.385 ng/ml and 0.107 ng/ml and quantification
limits were found to be 7.228ng/ml and 0.325ng/ml for RIFA and PIPE, respectively.
The method was validated for accuracy, precision, reproducibility, specificity, robustness,
and detection and quantification limits, in accordance with ICH guidelines.
Conclusion: Stress study was performed on RIFA and PIPE and it was found that these
degraded sufficiently in all applied chemical and physical conditions. Thus, the developed
RP-HPLC method was found to be suitable for the determination of both the drugs
in bulk as well as stability samples of capsule containing various excipients.