High-performance liquid chromatographic analysis of Black Cohosh (Cimicifuga racemosa) constituents with in-line evaporative light scattering and photodiode array detection Original Research Article

A validated and reproducible high-performance liquid chromatography (HPLC) method with in-line evaporative light scattering and photodiode array detection was developed for the analysis of major constituents in Black Cohosh (Cimicifuga racemosa L.). The method is based on the baseline chromatographic separation of 18 compounds reported to be present in Black Cohosh on a C-18 column (5 μm, 4.6 mm×250 mm) with water (0.05% trifluoroacetic acid, TFA)–acetonitrile–water as the mobile phase, and their in-line detection using photodiode array detector (PDA) and evaporative light scattering detector (ELSD). Sixteen of these (caffeic acid, ferulic acid, isoferulic acid, cimicifugoside H-1, cimiracemoside A, cimicifugoside H-2, (26R)-actein, 26-deoxycimicifugoside, (26S)-actein, 23-epi-26-deoxyactein, 23-OAc-shengmanol-3-O-β-d-xyloside, 26-deoxyactein, 25-OAc-cimigenol-3-O-α-l-arabinoside, 25-OAc-cimigenol-3-O-β-d-xyloside, cimigenol-3-O-α-l-arabinoside, cimigenol-3-O-β-d-xyloside) were detected to be present and quantifiable. The reputed two flavonoid constituents (kaempferol and formononetin) were not detectable.