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Planta Med 2017; 83(10): 855-861
DOI: 10.1055/s-0043-102689
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

Tharita Kitisripanya
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Supaluk Krittanai
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Orapin Udomsin
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Kamonthip Jutathis
3   Regional Medical Sciences Center 7, Department of Medical Sciences, Khon Kaen, Thailand
,
Jukrapun Komaikul
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Thaweesak Juengwatanatrakul
4   Faculty of Pharmaceutical Sciences, UbonRatchathani University, Ubon Ratchathani, Thailand
,
Seiichi Sakamoto
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Hiroyuki Tanaka
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Satoshi Morimoto
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Waraporn Putalun
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
› Author Affiliations
Further Information

Publication History

received 31 October 2016
revised 29 December 2016

accepted 25 January 2017

Publication Date:
10 February 2017 (online)

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Abstract

Miroestrol is a chromene with potent estrogenic activity present in Pueraria candollei, commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of P. candollei. The developed ELISA against miroestrol has a calibration range of 10–780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of P. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

Key words

miroestrol - monoclonal antibody - enzyme-linked immunosorbent assay (ELISA) - immunoaffinity column - White Kwao Krua - Pueraria candollei - Fabaceae
 

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