Development and Validation of High-Performance Thin-layer Chromatographic Method for Quantification of Berberine in Rhizomes of Coptis teeta Wall, an Endangered Species Collected from Arunachal Pradesh, India

The high polarity of the protoberberine alkaloids present in Coptis teeta has made it difficult to quantify the alkaloids. This study was designed to develop and validate a thin-layer chromatography (TLC) densitometric-based method using high-performance thin-layer chromatography for quantification of berberine. The separation was achieved in a solvent system consisting of butanol: ethyl acetate: formic acid: water by volume on TLC aluminum plate precoated with silica gel 60 F-254. Determination and quantification were performed by densitometric scanning under mercury lamp at a wavelength 351 nm in absorbance mode. The validated method gave compact bands for berberine at an R-f of 0.70. The precision, accuracy and reproducibility of the method were validated by following International Conference on Harmonization guidelines. Graphically, linear results were obtained for berberine with correlation coefficient of 0.997 +/- 0.09% (R +/- SD) in the concentration range of 90-210 ng/band. The limit of quantification and limit of detection from the analysis were found to be 70 and 30 ng/band, respectively. The berberine concentration in the methanol extract of C. teeta was found to be 30.97 +/- 0.55 mg in 100 mg of the crude drug. The method developed here in can be implemented in the analysis and routine quality control of herbal materials and formulations containing C. teeta and berberine.