Development and Validation of HPLC Method for Analysis of Picroside-I and Picroside-II in Picrorhiza kurroa

Aim: In the present investigation, our aim is to develop and validate HPLC method as per ICH guidelines for analyisis of picroside-I and picroside-II in Picrorhiza kurroa.

Place and Duration of Study: Investigation was undertaken in Department of Forest Products, University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India and in the period between June 2016 and December 2016.

Methodology: The system used is of Waters binary HPLC unit with Waters HPLC pump 515, dual λ absorbance detector 2487 and Empower II software. Standards of picroside-I and picroside-II were purchased and used for HPLC method development and validation. The developed HPLC method was validated for parameters as linearity, range, accuracy, precision, LOD and LOQ as mentioned in ICH guidelines.

Results: The analytical column, Sunfire C18 (4.6 × 250 mm, 5 µm) was operated at ambient temperature. Isocratic elution with A methanol and B water (40:60, v/v) at a flow rate of 0.9ml/min was selected. UV detection was done at 270 nm and run time was given forty minutes for standard compounds and forty five minutes for samples of Picrorhiza kurroa.

Conclusion: Method was found to be satisfactory in terms of linearity, high accuracy and precision. The method was successfully applied to the extracts made of different market samples of Picrorhiza kurroa.