Enzyme-Linked Immunosorbent Assay for Total Isoflavonoids in Pueraria candollei Using Anti-Puerarin and Anti-Daidzin Polyclonal Antibodies

Pueraria candollei (White Kwao Khuer) is amedicinal plant containing puerarin, daidzin, genistin, daidzein, and genistein as major isoflavonoids used for its rejuvenating and estrogenic effects. In order to analyze these compounds, a single enzyme-linked immunosorbent assay (ELISA) for total isoflavonoids was developed using anti-puerarin and anti-daidzin polyclonal antibodies (PAbs). The range for calibration of isoflavonoids by ELISA was 0.05–6.25 μg/mL. Total isoflavonoid concentrations in P. candollei samples determined by the newly developed assay system showed good agreement with those analyzed by HPLC. Based on validation analysis, this analytical method by ELISA is a precise, accurate, and sensitive method for the determination of total isoflavonoids in P. candollei.

Benyakan Pongkitwitoon, Seiichi Sakamoto, Hiroyuki Tanaka, Ryota Tsuchihashi, Junei Kinjo, Satoshi Morimoto and Waraporn Putalun
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan

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