Laboratory of Pharmacognosy and Phytochemistry, Ghent University-UGent, Harelbekestraat 72, B-9000 Gent, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Apr 15;878(13-14):949-56. doi: 10.1016/j.jchromb.2010.02.022. Epub 2010 Feb 24.
The investigation into the potential usefulness of phytoestrogens in the treatment of menopausal symptoms requires large-scale clinical trials that involve rapid, validated assays for the characterization and quantification of the phytoestrogenic precursors and their metabolites in biological matrices, as large interindividual differences in metabolism and bioavailability have been reported. Consequently, a new sensitive high-performance liquid chromatography-mass spectrometry method (HPLC-MS) for the quantitative determination of thirteen phytoestrogens including their most important gut microbial metabolites (genistein, daidzein, equol, dihydrodaidzein, O-desmethylangolensin, coumestrol, secoisolariciresinol, matairesinol, enterodiol, enterolactone, isoxanthohumol, xanthohumol and 8-prenylnaringenin) in human urine and serum within one single analytical run was developed. The method uses a simple sample preparation procedure consisting of enzymatic deconjugation followed by liquid-liquid extraction (LLE) or solid-phase extraction (SPE) for urine or serum, respectively. The phytoestrogens and their metabolites are detected with a single quadrupole mass spectrometer using atmospheric pressure chemical ionization (APCI), operating both in the positive and the negative mode. This bioanalytical method has been fully validated and proved to allow an accurate and precise quantification of the targeted phytoestrogens and their metabolites covering the lower parts-per-billion range for the measurement of relevant urine and serum levels following ingestion of phytoestrogen-rich dietary supplements.
植物雌激素在治疗更年期症状中的潜在用途的研究需要大规模的临床试验，这些试验需要快速、经过验证的分析方法来对生物基质中的植物雌激素前体及其代谢物进行特征描述和定量，因为据报道，个体间的代谢和生物利用度存在很大差异。因此，开发了一种新的灵敏高效液相色谱-质谱法（HPLC-MS），用于定量测定 13 种植物雌激素及其最重要的肠道微生物代谢物（染料木黄酮、大豆苷元、黄豆黄素、大豆苷元、O-去甲基安格洛宁、黄豆黄素、开环异落叶松脂醇、开环异落叶松脂醇、肠二醇、肠内酯、异黄腐醇、黄腐醇和 8-异戊烯基柚皮素）在人尿和血清中的含量，可在一次分析运行中完成。该方法采用简单的样品前处理程序，包括酶解共轭，然后分别进行尿液的液-液萃取（LLE）或血清的固相萃取（SPE）。植物雌激素及其代谢物采用大气压化学电离（APCI），在正离子和负离子模式下，用单四极杆质谱仪进行检测。该生物分析方法已得到充分验证，并证明能够准确、精密地定量测定目标植物雌激素及其代谢物，涵盖了摄入富含植物雌激素的膳食补充剂后相关尿液和血清水平的测量下限部分。
