Quantitative Determination of Three Protoberberine Alkaloids in Jin-Guo-Lan by HPLC-DAD

An accurate and simple HPLC method for the simultaneous determination of three protoberberine alkaloids (columbamine, jatrorrhizine, and palmatine) contained in Chinese medicine Jin-Guo-Lan (Tinospora sagittata Oliv. and Tinospora capillipes Gagnep) is presented in this study. The herb samples from six main origins and three herb markets were investigated. The separation was performed on a YMC-C₁₈ ODS column at 30°C with a gradient elution program. Acetonitrile and phosphate buffer (0.02 mol L⁻¹ sodium dihydrogen phosphate and 0.01 mol L⁻¹ triethylamine, pH 3) were used as mobile phases and the flow rate was set at 1 mL min⁻¹. The recovery of the method was in the range of 99.43–100.96%, and all the alkaloids showed good linearity (r ² > 0.9997) in the relatively wide concentration ranges. The developed method was applied to the determination of these alkaloids in the collected herb samples, and the results showed that the contents of these components in Jin-Guo-Lan varied greatly from habitat to habitat. It was demonstrated that the proposed method was helpful for the quality evaluation of Chinese medicine Jin-Guo-Lan.