ABSTRACT

Deoxymiroestrol is the most potent phytoestrogen in the chromene group found in Pueraria candollei. The estrogenic activity of P. candollei, functions as hormone replacement therapy in postmenopausal women. This study established an enzyme-linked immunosorbent assay (ELISA) for quantifying deoxymiroestrol using fragment antigen-binding (Fab) antibody to improve analytical performance. Our ELISA specifically detected deoxymiroestrol in the range of 31.25–1,000 ng/mL. Intra- and inter-assay precision was 1.48%–7.11% and 0.58%–9.31%, respectively, the recovery of authentic deoxymiroestrol spiked into samples was 99.77%–101.61%, and the limit of detection was 30.80 ng/mL. The findings of P. candollei root bark determined using our developed ELISA with Fab antibody and an ELISA with anti-deoxymiroestrol polyclonal antibody were consistent (R² = 0.9807). The developed ELISA can be applied to the quality control of deoxymiroestrol derived from P. candollei.

摘要

Deoxymiroestrol 是在Pueraria candollei 中发现的色烯组中最有效的植物雌激素。P. candollei的雌激素活性在绝经后妇女中起到激素替代疗法的作用。本研究建立了一种酶联免疫吸附测定 (ELISA)，用于使用片段抗原结合 (Fab) 抗体对脱氧微雌酚进行定量，以提高分析性能。我们的 ELISA 专门检测了 31.25–1,000 ng/mL 范围内的脱氧微雌醇。批内和批间精密度分别为 1.48%–7.11% 和 0.58%–9.31%，掺入样品的正宗脱氧微雌酚的回收率为 99.77%–101.61%，检出限为 30.80 ng/mL。P. candollei的发现使用我们开发的使用 Fab 抗体的 ELISA 和使用抗脱氧微雌醇多克隆抗体的 ELISA 测定的根皮根皮是一致的 (R ²  = 0.9807)。开发的ELISA可用于P. candollei来源的脱氧微雌酚的质量控制。