Stability Indicating Simultaneous Quantification of Chlorogenic Acid and Berberine in Homeopathic Polyherbal Formulation by AQbD Based HPLC

Background: Analytical Quality by Design (AQbD) enhances method robustness more effectively than traditional methods. Risk assessment and factor screening studies enabled to detection of analytical method parameters. The present study describes an AQbD enabled stability indicating RP-HPLC method for simultaneous determination of the phytochemical markers, Chlorogenic acid and Berberine in standard and clear stone drops, a Homeopathic marketed formulation. Currently no published methods reported based on stability indicating RP-HPLC technique for determination of these markers in any herbal formulations. Materials and Methods: Two factors at three levels were considered using Central Composite Design for the method optimization. Statistical and graphical analyses were employed to assess the individual and combined interaction effects of critical method parameters on critical method responses. Results: The optimized mobile phase was Methanol: 0.1% Formic acid (41.67:58.33% v/v) with 1.1 mL/min rate of flow, at 343 nm. Chlorogenic acid was eluted at 3.534 min and Berberine was eluted at 5.140 min respectively with 8 min run time. The method was validated as per ICH Q2 R1 guidelines and performed stress degradation studies; all validation parameters were within acceptable limits. Conclusion: The established HPLC technique was useful as quality control tool for assessment of phytochemical markers in various traditional medicines and successfully applied to determine Chlorogenic acid and Berberine in Marketed Homeopathic formulation.