Enzyme-Linked Immunosorbent Assay for Determination of Miroestrol in Pueraria candollei Associated Products

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Gorawit Yusakul
Orapin Udomsin
Thaweesak Juengwatanatrakul
Hiroyuki Tanaka
Chaiyo Chaichantipyuth
Waraporn Putalun


Introduction: Various dosage forms of Pueraria candollei are widely applied in folk Thai medicine for rejuvenating purpose in aged people, which correlated with its pharmacological activities reported by pre-clinical and clinical trials for menopausal symptoms. Therefore, standardized products of this plant are needed by consumers and health care personnel.  Miroestrol (ME), a potent and stable phytoestrogen in P. candollei, exhibited potential to be biomarker for quality control of P. candollei samples and its products. Materials and methods: Indirect competitive enzyme-linked immunosorbant assay (ELISA) for ME determination was developed and validated by using polyclonal antibody from rabbit immunization. The correlation of ME determination between our ELISA with standard HPLC method was investigated. Finally validated ELISA was applied for P. candollei associated products. Results: Developed ELISA was comparable with published HPLC method (R2= 0.9996) in samples with various ME contents, moreover this ELISA could determine ME in samples undetectable by HPLC because of its higher sensitivity. For application, the P. candollei involved preparations contained ME of 0.766- 12.108 µg/g dry wt. Conclusion: The developed ELISA was high performance for reliable ME determination, which could be applied for P. candollei quality control in research fields and industrial productions.


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