Determination of Seven Biflavones of Selaginella Doederleinii by High Performance Liquid Chromatography

Abstract

Selaginella doederleinii Hieron (S. doederleinii) is widely used as an antitumor and anti-inflammatory agent in Asia, and is famous for containing biflavonoids, which have been reported as the bio-active constituents possessing anticancer and anti-oxidant effects. However, a simple and systematic quality control method for determining biflavonoids in S. doederleinii is lacking. In this study, seven biflavones, including amentoflavone, robustaflavone, 2″, 3″-dihydro-3′, 3″′-biapigenin; 3′, 3″′-binaringenin, delicaflavone, heveaflavone, and 7, 4′, 7″, 4″′-tetra-O-methyl-amentoflavone, were obtained from S. doederleinii. These seven compounds that are present in high concentration were used as detection markers for development of a quality control method by HPLC with a UV-visible detector. The chromatographic separation was accomplished on a C18 column with a mobile phase composed of acetonitrile–water (0.5% acetic acid) in 52 min. With the optimized conditions, the seven biflavones showed good regression relationships (R2 > 0.9989) within the test range and the recovery was in the range of 90.1–102.5%. The LODs of the seven analytes were between 0.2 and 1.17 µg/mL. The method was also validated for precision and accuracy and was successfully applied to the determination of these compounds in six batches of S. doederleinii from different areas. In conclusion, the present study provides a systematic and useful method for quality evaluation of S. doederleinii.