LC Determination of Four Isoflavone Aglycones in Red Clover (Trifolium pratense L.) |
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Authors: | Graziele P Ramos Paula M B Dias Cláudia B Morais Pedro E Fröehlich Miguel Dall’Agnol José A S Zuanazzi |
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Institution: | 1. Programa de Pós-Gradua??o em Ciência Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga, 2752, CEP 90.610-000, Porto Alegre, RS, Brazil 3. Programa de Pós-Gradua??o em Zootecnia, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gon?alves, 7712, CEP 91.501-970, Porto Alegre, RS, Brazil 2. Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga, 2752, CEP 90.610-000, Porto Alegre, RS, Brazil
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Abstract: | Red clover (Trifolium pratense L.) is an important forage plant that contains the isoflavones daidzein, genistein, formononetin, and biochanin A. These
compounds have been studied lately due to their human health benefits. The aim of this study was to develop and validate an
HPLC method with simplified sample preparation to quantify daidzein, genistein, formononetin and biochanin A simultaneously
in red clover leaves. The validation showed that the method is specific, accurate, precise and robust, not to mention that
the sample preparation is easier and faster than those described earlier. The response was linear over a range of 0.01–0.2 μg mL−1 for daidzein, 0.05–0.5 μg mL−1 for genistein, 4–40 μg mL−1 for formononetin and 2–20 μg mL−1 for biochanin A. The range of recoveries was 85.6–101.0%. The RSD for intra- and inter-day precision were <2.54 and <7.22%,
respectively. Five populations of red clover, from the National Plant Germplasm System-USDA were analyzed and the content
of daidzein, genistein, formononetin and biochanin A ranged from 7.87–91.31, 51.60–131.30, 6568.33–23461.82, to 2499.55–10337.33 μg g−1 of dried material, respectively. |
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Keywords: | Column liquid chromatography Red clover Isoflavones Validation |
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