Development of validated chemical fingerprint for an Ayurvedic formulation Kantakaryaavaleha

Kantakaryaavaleha is a most trusted Ayurvedic formulation among the Ayurvedic medicines. Kantakaryaavaleha is mentioned throughout the ancient literature of Ayurvedic medicine and used for svasa (asthma), kassa (cough), sula (colic). A simple, sensitive, selective, precise and robust high-performance thin-layer chromatographic (HPTLC) method was developed and validated for the determination and routine analysis of plumbagin in a Traditional Indian formulation (Kantakaryaavaleha) and its crude drug extracts. The validation parameter of developed HPTLC method was found to be reproducible. Analysis was performed on TLC aluminum plates pre-coated with silica gel 60F-254 as the stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with mobile phase consisting of toluene: ethyl acetate: MeOH (8:1:1). Camag TLC scanner III was used for spectro densitometric scanning and analysis in absorbance mode at 265nm. The system was found to give compact spots for plumbagin (Rf value of 0.49 ± 0.02). The HPTLC linear regression analysis data for the calibration plots showed good linear relationship (r²= 0.9998 ± 0.0003) in the concentration range 200 – 600 ng spot^–1 with respect to peak area, moreover the method was validated for precision, recovery, robustness and ruggedness according to the International Conference on Harmonization (ICH) guidelines. The plumbagin content was quantified and estimated from the formulation and the plumbago zeylanica plant part. Statistical analysis of the data showed that the method is reproducible and selective for the quantitative determination of plumbagin.