Five major flavonoids in 48 batches of Radix Hedysari from different origins have been simultaneously evaluated by HPLC. Among the five major flavonoids, four, ononin, formononetin-7-O-β-D-glucopyranoside-6″-O-malonate, formononetin and medicarpin, were detected in almost all the samples and thus can be used as marker compounds to evaluate the chemical quality of Radix Hedysari, while naringin was not suitable because it was not be detected in most samples. Further analysis of the contents of the four flavonoids in different samples showed that processing procedure, harvesting time and habitats were important factors affecting the flavonoid contents of Radix Hedysari.
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