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 ¹H-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.

由于葛根（PM）的保健食品偶尔会对健康造成损害，日本政府将其指定为“需要特别注意的成分”。Miroestrol 是一种从 PM 中分离出来的特定异黄酮，具有很强的雌激素活性，足以引起少量副作用。因此，建议对微雌酚定量进行常规分析，以控制 PM 产品的安全性和质量。然而，PM中的微雌酚含量很低，并且很少有用于其检测的商业试剂。在本研究中，我们通过使用相对摩尔灵敏度（RMS) 的 miroestrol 到 kwakhurin，另一种 PM 特异性异黄酮，作为参考标准。的RMS是由脱机组合获得的值¹ H-NMR定量光谱和一个LC /光电二极管阵列（PDA）和葛雌素含量通过单引用LC / PDA使用确定RMS。此外，我们调查了市售 PM 生药和产品中的微雌酚含量，并在性能方面将RMS方法与常规校准曲线方法进行了比较。两种方法测定的微雌激素含量一致率为89-101%。结果表明，我们开发的 LC/PDA/MS 方法与RMS 以kwakhurin为参考标准，可准确监测PM生药及制品中微雌酚含量的常规监测，以控制其质量。