The determination of coumestrol in alfalfa (Medicago sativa) by capillary electrophoresis
J. Moravcová, T. Kleinová, R. Loučkahttps://doi.org/10.17221/4230-PSECitation:Moravcová J., Kleinová T., Loučka R. (2002): The determination of coumestrol in alfalfa (Medicago sativa) by capillary electrophoresis. Plant Soil Environ., 48: 224-229.
High performance capillary electrophoresis (HPCE) on an uncoated fused-silica capillary column using a borate buffer at pH 9.2 as electrolyte and diode-array detection was developed for the determination of coumestrol in alfalfa. The linear detector response was established in the concentration range 0.76–140 mg.dm–3, the minimum detectable limit was 0.39 mg.dm–3, and migration time of coumestrol was 5 min. 3-Isobutyl-1-methylxanthin was used as an internal standard. Coumestrol was isolated by acid extraction employing a mixture hydrochloric acid-acetonitrile at 95°C for 30 min followed by solid phase extraction. Relative standard deviations of reproducibility and repeatability were 1.77% and 5.49%, respectively. Spiking recovery value of 92% was achieved. Alfalfa, variety Morava, contains 148–248 mg.kg–1 coumestrol in dry matter. The proposed method is useful for routine analyses.Keywords:
phytoestrogens; alfalfa; plant analysis; animal feed