Luísa Ma Martin

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José Mário Girão Abreu

Universidade Estadual do Ceará

Ma. Conceição Castilho

University of Coimbra

M.I.N. Silveira

University of Coimbra

Cooperative Institutions

University of Coimbra

Universidade Estadual do Ceará

Universidade Federal de Viçosa

Universidade Federal Rural de Pernambuco

Universidade Federal de Campina Grande

Universidade de Fortaleza

Centro de Estudos e Sistemas Avançados do Recife

University of Trás-os-Montes and Alto Douro

Charles University

Universitat de València

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Liquid Chromatographic Validation of a Quantitation Method for Phytoestrogens, Biochanin‐A, Coumestrol, Daidzein, Formononetin, and Genistein, in Lucerne

Journal of Liquid Chromatography &amp Related Technologies (2006)

Luísa Ma Martin Ma. Conceição Castilho M.I.N. Silveira José Mário Girão Abreu

Abstract A methodology was validated to evaluate biochanin‐A, coumestrol, daidzein, formononetin, and genistein in lucerne (Medicago sativa). Extraction done through acid hydrolysis at 80°C, followed by extraction with HLB cartridges. A C18 column (4.6×250 mm) with a 0.8 mL/min flow of methanol solution (isocratic) and detection at 260 nm was applied. Separation factors were greater than 1.1 and resolutions were superior to 2.4. Linearity was established and correlation coefficients of responses were greater than 0.995. The detection and quantification limits were between 0.12–0.17 µg · mL−1 and 0.37–0.53 µg · mL−1, respectively. The coefficients of variation were between 5 and 15%. The accurate results in lucerne were 78, 35, 20, 14, and 11 mg (kgDM)−1 for coumestrol, formononetin, daidzein, genistein, and biochanin‐A.

Biochanin A

Coumestrol

10.1080/10826070600961076

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