Journal
CHEMICAL & PHARMACEUTICAL BULLETIN
Volume 69, Issue 6, Pages 573-580Publisher
PHARMACEUTICAL SOC JAPAN
DOI: 10.1248/cpb.c21-00160
Keywords
Pueraria mirifica; quality control; miroestrol; kwakhurin; relative molar sensitivity; quantitative NMR
Funding
- Ministry of Health, Labour, and Welfare of Japan
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By developing a quantitative analysis method for miroestrol in Pueraria mirifica (PM) products using kwakhurin as a reference standard, this study successfully monitored the accuracy of miroestrol content in PM products. The results showed that the developed LC/PDA/MS method with RMS was accurate for routine monitoring of miroestrol content in PM products to control their quality.
Owing to occasional health damages caused by health food products derived from Pueraria mirifica (PM), the Japanese government has designated PM as an ingredient calling for special attention. Miroestrol is a specific isoflavone isolated from PM and possesses very strong estrogenic activity enough to induce side effects in small amount. Therefore, routine analyses for miroestrol quantification is recommended to control the safety and quality of PM products. However, miroestrol content in PM is quite low, and commercial reagent for its detection is rarely available. In this study, we developed a quantitative analysis method for miroestrol in PM without using its analytical standard by using the relative molar sensitivity (RMS) of miroestrol to kwakhurin, another PM-specific isoflavone, as a reference standard. The RMS value was obtained by an offline combination of H-1-quantitative NMR spectroscopy and a LC/photo diode array (PDA) and miroestrol content was determined by single-reference LC/PDA using RMS. Furthermore, we investigated miroestrol content in commercially available PM crude drugs and products, and the RMS method was compared with the conventional calibration curve method in terms of performance. The rate of concordance of miroestrol contents determined by two method was 89-101%. The results revealed that our developed LC/PDA/MS method with RMS using kwakhurin as a reference standard was accurate for routine monitoring of miroestrol content in PM crude drugs and products to control their quality.
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