Pan Jian-Yang, Zhao Fang, Li Wen-Zhu, Qu Hai-Bin
Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences,Zhejiang University Hangzhou 310058, China Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine Hangzhou 310058, China.
Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences,Zhejiang University Hangzhou 310058, China Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine Hangzhou 310058, China Hisun Biopharmaceutical Co., Ltd. Hangzhou 311400, China.
Zhongguo Zhong Yao Za Zhi. 2022 Feb;47(3):575-580. doi: 10.19540/j.cnki.cjcmm.20211108.304.
The present study established a quality evaluation method for ginsenoside reference substances based on quantitative nuclear magnetic resonance(qNMR) spectroscopy. ~1H-NMR spectra were collected on Bruker Avance Ⅲ 500 MHz NMR spectrometer equipped with a 5 mm BBO probe. The acquire parameters were set up as follows: pulse sequence of 30°, D_1=20 s, probe temperature= 303 K, and the scan number = 32. Dimethyl terephthalate, a high-quality ~1H-qNMR standard, was used as the internal standard and measured by the absolute quantitative method. Methyl peaks of comparatively good sensitivity were selected for quantification, and linear fitting deconvolution was adopted to improve the accuracy of integration results. The qNMR spectroscopy-based method was established and validated, which was then used for the quality evaluation of ginsenoside Rg_1, ginsenoside Re, ginsenoside Rb_1, ginsenoside Rd, and notoginsenoside R_1. The results suggested that the content of these ginsenoside reference standards obtained from the qNMR spectroscopy-based method was lower than that detected by the normalization method in HPLC provided by the manufacturers. In conclusion, the qNMR spectroscopy-based method can ensure the quality of ginsenoside reference substances and provide powerful support for the accurate quality evaluation of Chinese medicine and its preparations. The qNMR spectroscopy-based method is simple, rapid, and accurate, which can be developed for the quantitative assay of Chinese medicine standard references.
本研究建立了基于定量核磁共振（qNMR）光谱法的人参皂苷对照品质量评价方法。在配备5 mm BBO探头的布鲁克AvanceⅢ 500 MHz核磁共振仪上采集1H-NMR光谱。采集参数设置如下：脉冲序列为30°，D1 = 20 s，探头温度 = 303 K，扫描次数 = 32。使用对苯二甲酸二甲酯这种高质量的1H-qNMR标准品作为内标，并采用绝对定量法进行测定。选择灵敏度相对较好的甲基峰进行定量，并采用线性拟合去卷积来提高积分结果的准确性。建立并验证了基于qNMR光谱法的方法，然后将其用于人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd和三七皂苷R1的质量评价。结果表明，基于qNMR光谱法获得的这些人参皂苷对照品的含量低于制造商提供的HPLC归一化法所检测到的含量。总之，基于qNMR光谱法的方法能够确保人参皂苷对照品的质量，并为中药及其制剂的准确质量评价提供有力支持。基于qNMR光谱法的方法简单、快速且准确，可用于开发中药标准对照品的定量测定方法。
