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Abstract
A simple, sensitive, and reliable HPLC method was developed for the overall separation of different chemical constituents present in the rhizome of Polygonum bistorta. The extraction conditions including soaking, sonication for 30 minutes, and heating for one hour were selected to achieve complete extraction. Different chromatographic conditions such as mobile phase composition and pH were studied and compared in the Reversed Phase separation mode to acquire fingerprinting profiles. 30 distinct peaks were obtained and 3 phenolic acids (gallic acid, protocatechuic acid, and chlorogenic acid) with anticancer bioactivity were identified by comparing their retention times with reference marker compounds in the optimized method. The excellent repeatability was tested by calculating relative standard deviations (RSDs) for retention times and peak areas and resultant values were less than 0.5% and 1.2%, respectively, for these respective compounds. This method can be used for quality assessment and further identification, quantification, and purification of the important bioactive constituents present in the rhizome of Polygonum bistorta.
ACKNOWLEDGMENT
This work was kindly supported by “The 973 Project from the Ministry of Science and Technology of People's Republic of China (No. 2011CB910404)” and “The National Natural Science Foundation of China (No. 21075039/B050106)”.