TLC densitometric fingerprint development and validation of Berberine as markers in poly-herbal Unani formulations
By Shiv Shankar Shukla, Swarnlata Saraf, Shailendra Saraf
Sat-e-gilo (Tinospora cardifolia) and its poly herbal formulations have been in use in Indian traditional medicine since antiquity. However, there has been no attempt to standardize the polyherbal formulations containing Berberine as the main ingredient in terms of its active principal or marker compound. Biologically active Berberine is identified as the marker compound for the crude drugs and its formulations. In the present study, the method for fingerprints of the formulations in the form of densitogram following charring of the chromatographic plate was developed that could be useful for marker-based quality assurance of the polyherbal products containing Tinospora cardifolia. Accuracy was validated by analysis of spiked blank and standard addition samples and precision by performing replicate analyses on a single day and on different days. Recoveries from spiked blank and standard addition sample were 98.03-100.37%. Repeatability for sample, each of which was analyzed six times on a single plate, was 0.034967 % relative standard deviation. The intra and inter precision was 0.045 and 0.047% relative standard deviation for a sample analyzed in duplicate once per plate on same day and the different days over a seven-day period. The content of Berberine was found to be 11.75 ± 0.0208– 12.46 ± 0.0264 mg/gm in lab formulations. A survey was made of Berberine content compared with label values for two marketed products, using the new method. The method was shown to be simple, precise and accurate suitable for routine analysis of Berberine in a manufacturing quality-control or regulatory agency laboratory.
Keywords :Densitometry, HPTLC, Berberine, Unani formulation, Fingerprint, Giloe.