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

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Keywords:
P. candollei miroestrol enzyme-linked immunosorbant assay phytoestrogen polyclonal antibod

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Gorawit Yusakul
BPharm, Student, Master of Pharmacy in Pharmaceuticals, Faculty of Pharmaceutical Sciences, Khon Kaen University and Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Thailand.
Orapin Udomsin
BPharm, Student, Doctor of Philosophy in Pharmaceuticals, Faculty of Pharmaceutical Sciences, Khon Kaen University and Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Thailand.
Thaweesak Juengwatanatrakul
Ph.D., Assistant Professor, Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Ubon Rajathanee University.
Hiroyuki Tanaka
Ph.D., Associate Professor, Graduate School of Pharmaceutical Sciences, Kyushu University, Japan.
Chaiyo Chaichantipyuth
Ph.D., Assoc. Prof. Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Thailand.
Waraporn Putalun
Ph.D., Associate Professor, Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University and Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Thailand.

Abstract

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.

Article Details

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Vol. 9 No. 1 (2013): Volume 9 No.1 January – April 2013
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