A rapid and simple determination of protoberberine alkaloids in Rhizoma Coptidis by 1H NMR and its application for quality control of commercial prescriptions

Abstract

Simple, convenient, sensitive and accurate analytical methods are needed for the analysis of alkaloid components in Rhizoma Coptidis in traditional Chinese herbal medicine, which has important bioactivity. In the present study, a highly specific and sensitive method using 1H NMR has been developed for the quantitative determination of protoberberine alkaloids berberine, palmatine, coptisine and jatrorrhizine in Coptis species and their commercial traditional Chinese medicine prescriptions. A 1H NMR analysis of the H-13 signals of target protoberberine alkaloids was performed. By comparing the solvent effects on the resolution of these signals, methanol-d4–benzene-d6 (75:25) is selected as an optimal 1H NMR solvent. The quantity of the compounds is calculated by the relative ratio of the integral values of the target peak for each compound to the known amount of the internal standard anthracene. This method allows rapid and simple quantitation of protoberberine alkaloids from Coptis species and the more complex commercial prescriptions in 5min without any pre-purification steps. The recoveries of these alkaloids from Coptis chinensis are in the range of 93–105%. Limit of detection of berberine in the plant material or prescription is 0.03mg/mL. The advantages of this method are that no reference compounds are required for calibration curves, the quantification can be directly realized on a crude extract, and the better selectivity for protoberberine alkaloids and a very significant time-gain can be achieved, in comparison to conventional HPLC methods, for instance.