Miroestrol is potent phyotoestrogen from Pueraria candollei tuberous root. Enhanced chemiluminescence enzyme linked immunosorbent assay (ECLELISA) was developed and validated using polyclonal antibody against miroestrol and chemiluminescence system of luminol-H2O2-horseradish peroxidase-4-(1-imidazolyl) phenol. The ECL-ELISA system exhibited the linearity of 0.31 – 10.00 ng/ml, which the relative standard variation (% RSD) are less than 10% of both intra- and interplate determinations. The ECL-ELISA is reliable to determine miroestrol reflected by high percentage of recovery (101.22 – 103.06%). As comparative analysis, ME contents in each sample determined by ECL-ELISA were correlated with high coefficient of determination to colorimetric ELISA (R2= 0.998) and HPLC method (R2=0.998). This method could be applied to all sampled of P. candollei root involved commercial products, which theses products contained 0.71 – 13.12 μg/g dry wt. of miroestrol. Totally, this method is useful as high performance analytical method for miroestrol quantity control in raw material and its product for both research and industrial levels.
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