Journal of Natural Medicines ( IF 2.5 ) Pub Date : 2021-06-09 , DOI: 10.1007/s11418-021-01539-5 Worapol Sae-Foo 1 , Supaluk Krittanai 1 , Wipawee Juengsanguanpornsuk 1 , Gorawit Yusakul 2 , Seiichi Sakamoto 3 , Waraporn Putalun 1
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Pueraria candollei is an ingredient of Thai herbal medicine, dietary supplements, and cosmetics. The in vitro and in vivo studies of this plant supported anti-osteoporotic activity and used for hormone replacement therapy. Deoxymiroestrol shows the most potent phytoconstituent in tuberous root of P. candollei with estrogenic activity. The quality controls are important for good agricultural practice (GAP) and good manufacturing practice (GMP) of plant-derived raw materials. The rapid detection of lateral flow immunoassay (LFIA) using colloidal gold is simply method, easy visualize detection and produce less waste than conventional chromatographic detection. In this study, LFIA for qualitative detection of deoxymiroestrol using antigen-binding fragment antibody (Fab) was developed. The result showed that the developed LFIA displays specific detection of deoxymiroestrol. Cross reactivity of this method was analyzed with miroestrol, isomiroestrol and methylisomiroestrol which showed 39.97%, 7.71% and 5.72%, respectively. After optimal condition, limit of detection (LOD) for deoxymiroestrol is 250 ng/ml. Plant samples were applied to strip test compare with indirect competitive ELISA using polyclonal antibody to confirm the application of LFIA. The results of LFIA method were comparable with those from ELISA. This developed lateral flow immunoassay can apply to detect deoxymiroestrol for the rapid testing. The developed method can use for quality control in plant samples as deoxymiroestrol is biomarker compound in P. candollei.
Graphic abstract
中文翻译：
基于片段抗原结合 (Fab) 抗体的侧向流动免疫测定法，可快速灵敏地检测强效植物雌激素、脱氧微雌激素
Pueraria candollei是泰国草药、膳食补充剂和化妆品的一种成分。该植物的体外和体内研究支持抗骨质疏松活性并用于激素替代疗法。Deoxymiroestrol 显示出具有雌激素活性的P. candollei块根中最有效的植物成分。质量控制对于植物源性原材料的良好农业规范 (GAP) 和良好生产规范 (GMP) 很重要。使用胶体金的横向流动免疫分析 (LFIA) 快速检测方法简单，检测容易可视化，比传统色谱检测产生更少的浪费。在本研究中，开发了使用抗原结合片段抗体 (Fab) 定性检测脱氧微雌酚的 LFIA。结果表明，开发的 LFIA 显示出脱氧微雌醇的特异性检测。该方法的交叉反应性与微雌醇、异异雌醇和甲基异异雌醇进行了分析，分别显示 39.97%、7.71% 和 5.72%。在最佳条件下，脱氧微雌醇的检测限 (LOD) 为 250 ng/ml。将植物样品用于条带试验，与使用多克隆抗体的间接竞争ELISA 进行比较，以确认LFIA 的应用。LFIA 方法的结果与 ELISA 方法的结果相当。这种开发的侧流免疫测定法可用于检测脱氧微雌酚以进行快速检测。所开发的方法可用于植物样品的质量控制，因为脱氧米罗雌酚是植物中的生物标志物化合物。P. 坎多莱。
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