Enzyme linked immunosorbent assay for total potent estrogenic miroestrol and deoxymiroestrol of Pueraria candollei, a Thai herb for menopause remedy.

Miroestrol and deoxymiroestrol are the most potent phytoestrogens of Pueraria candollei var. mirifica, having been proved as an effective herb for menopausal symptoms in folk medicines and clinical trials. To ensure efficacy and safety of P. candollei var. mirifica involved in nutraceutical products being available worldwide, the content of potent phytoestrogens as active ingredients should be specified. Therefore, in this study, we produced a monoclonal antibody for total analysis of potent estrogenic miroestrol and deoxymiroestrol, for which an analytical method was developed using a procedure for an indirect competitive enzyme-linked immunosorbent assay. The antibody exhibited equal reactivity against miroestrol and deoxymiroestrol. The sensitivity of determination was in the range of 31.3-500 ng/ml with high precision. The analytical parameters, such as accuracy (99.6-106% recovery) and high correlation with a HPLC-UV method, indicated the reliability of analysis. This method is of high performance, and it is cheap to control optimal miroestrol and deoxymiroestrol doses of P. candollei var. mirifica nutraceutical products.