Agroscope Reckenholz-Tänikon Research Station ART, CH-8046 Zürich, Switzerland.
J Agric Food Chem. 2011 Feb 9;59(3):847-56. doi: 10.1021/jf1039266. Epub 2011 Jan 4.
We developed and validated three different sample preparation and extraction methods followed by HPLC-MS/MS (negative electrospray ionization) analysis for the quantification of estrogenic isoflavones (formononetin, daidzein, equol, biochanin A, and genistein) and coumestrol in red clover, soil, and manure. Plant and manure samples were solid-liquid extracted, whereas soil was extracted with accelerated solvent extraction. Absolute recoveries were between 80 and 93%, 20 and 30%, and 14 and 91% for plant, soil, and manure samples, respectively. Relative recoveries ranged from 75 to 105% for all matrices, indicating that isotope-labeled internal standards (¹³C₃-formononetin, ¹³C₃-daidzein, ¹³C₃-equol, ¹³C₃-biochanin A, and ¹³C₃-genistein) were capable to compensate for losses during analysis. The limits of detection in red clover, soil, and manure were 3-9 μg/g(dryweight(dw)), 0.6-8.2 ng/g(dw), and 34.2 ng/g(dw) to 17.0 μg/g(dw), respectively. Formononetin was the most dominant compound in red clover plants (up to 12.5 mg/g(dw)) and soil (up to 3.3 μg/g(dw)), whereas equol prevailed in manure (up to 387 μg/g(dw)).
我们开发并验证了三种不同的样品前处理和提取方法，随后采用 HPLC-MS/MS（负离子电喷雾电离）分析测定了红三叶草、土壤和粪肥中雌激素异黄酮（芒柄花素、大豆苷元、黄豆黄素、染料木黄酮和染料木苷）和香豆雌酚的含量。植物和粪肥样品采用固液萃取，而土壤则采用加速溶剂萃取。植物、土壤和粪肥样品的绝对回收率分别为 80-93%、20-30%和 14-91%。所有基质的相对回收率范围为 75-105%，表明同位素标记的内标（¹³C₃-芒柄花素、¹³C₃-大豆苷元、¹³C₃-黄豆黄素、¹³C₃-染料木黄酮和¹³C₃-染料木苷）能够补偿分析过程中的损失。红三叶草、土壤和粪肥中的检测限分别为 3-9 μg/g（干重（dw））、0.6-8.2 ng/g（dw）和 34.2-17.0 μg/g（dw）。芒柄花素是红三叶草植物（高达 12.5 mg/g（dw））和土壤（高达 3.3 μg/g（dw））中含量最丰富的化合物，而黄豆黄素在粪肥中占主导地位（高达 387 μg/g（dw））。
