中国药物警戒 ›› 2025, Vol. 22 ›› Issue (6): 614-620.
DOI: 10.19803/j.1672-8629.20250078

• 基础与临床研究 • 上一篇    下一篇

远志药材及饮片中黄曲霉毒素含量测定方法优化研究

卢宇1,2, 冯伟红1, 程博宇1, 戴梦茹1, 卢晨娜1, 乔灵芝1, 张永欣1, 刘晓谦1, 梁曜华1, 李艳茸3, 孙晓丽2#, 李春1*   

  1. 1中国中医科学院中药研究所中药质量控制技术国家工程实验室,北京 100700;
    2北京城市学院生物医药学部,北京 100094;
    3榆林市第五医院药学部,陕西 榆林 719000
  • 收稿日期:2025-02-11 发布日期:2025-06-18
  • 通讯作者: *李春,女,博士,研究员,中药化学。E-mail: cli@icmm.ac.cn;#为共同通信作者。
  • 作者简介:卢宇,女,在读硕士,中药分析与质量标准研究。
  • 基金资助:
    国家重点研发计划（2023YFC3504000）; 中国中医科学院中药研究所“使命导向改革”重点专项（CI2023E001TS03）; 中国中医科学院科技创新工程（CI2021A04206、CI2021A04402、CI2021A04412）; 榆林市科技计划（YF-2021-74）

Optimization of a Determination Method for Aflatoxins in Polygalae Radix

LU Yu1,2, FENG Weihong1, CHENG Boyu1, DAI Mengru1, LU Chenna1, QIAO Lingzhi1, ZHANG Yongxin1, LIU Xiaoqian1, LIANG Yaohua1, LI Yanrong3, SUN Xiaoli2#, LI Chun1*   

  1. 1Institute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, National Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines, Beijing 100700, China;
    2Department of Biomedical Sciences, Beijing City University, Beijing 100094, China;
    3Department of Pharmacy, the Fifth Hospital of Yulin City, Yulin Shaanxi 719000, China
  • Received:2025-02-11 Published:2025-06-18

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摘要： 目的 建立准确测定远志药材和饮片中4种黄曲霉毒素的高效液相色谱（HPLC）方法。方法 采用Ultimate XB-C18色谱柱（4.6 mm×150 mm,5μm）,以甲醇-乙腈-水（22∶10∶68）为流动相,流速0.7 mL·min-1,柱温40℃,荧光检测器（λex=360 nm,λex=450 nm）检测,进样量10μL。对比不同提取方法与不同品牌免疫亲和柱的提取净化效果,优化前处理过程,考察基质效应对检测结果的影响,并对28批远志药材及饮片的黄曲霉毒素含量进行测定。结果 方法学考察结果符合中药质量分析要求。黄曲霉毒素G2（AFG2）、黄曲霉毒素B2（AFB2）在0.000 075~0.075 μg·mL-1,黄曲霉毒素G1（AFG1）、黄曲霉毒素B1（AFB1）在0.000 25~0.25 μg·mL-1内线性关系良好,相关系数均为0.999 9,基质效应为94.49%~103.51%,可视为远志基质对黄曲霉毒素的测定无影响。在低、中、高浓度下AFG2、AFG1、AFB2和AFB1的平均回收率分别为78.98%~93.51%、81.88%~100.74%、82.19%~116.12%,相对标准偏差（RSD）介于1.1%~6.6%。针对远志品种更适宜使用均质器提取黄曲霉毒素,以及免疫亲和柱富集。28批远志样品中,有2批药材不合格,其余26批药材与饮片均符合药典标准。黄曲霉毒素总量最高达313.63μg·kg-1,AFB1的含量最高达143.27μg·kg-1。结论 建立了远志药材中4种黄曲霉毒素同步测定的简便、可靠新方法,完善了已有检测方法,揭示了远志普遍存在黄曲霉毒素污染风险,为提升远志药材质量提供参考。

关键词: 远志, 黄曲霉毒素, 高效液相色谱-荧光检测联用, 基质效应

Abstract: Objective To establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of contents of four aflatoxins in Polygalae Radix medicinal materials. Methods An Ultimate XB-C18 column (4.6 mm×150 mm, 5 μm) was used as the stationary phase with methanol (A)-acetonitrile (B)-water (C) (22∶10∶68) as the mobile phase. The flow rate was 0.7 mL·min-1 and the column temperature 40°C. A fluorescence detector (λex=360 nm, λex=450 nm) was adopted, and the injection volume was 10 μL. The effects of extraction and purification were compared between different methods of extraction and between immunoaffinity columns of different brands. The pretreatment process was optimized, and the difference that matrix effects made in detection results was investigated. The aflatoxin levels of 28 batches of Polygalae Radix herbs and decoction pieces were tested. Results The results of the methodological investigation met the requirements of quality analysis of traditional Chinese medicine. AFG2 and AFB2 showed a good linear relationship within the range of 0.000 075 to 0.075 μg·mL-1, compared to 0.000 25 to 0.25 μg·mL-1 for AFG1 and AFB1.The correlation coefficients were 0.999 9. The matrix effect ranged from 94.49% to 103.51%, indicating that the Polygalae Radix matrix had no significant impact on the determination of aflatoxins. The average recovery of AFG2, AFG1, AFB2, and AFB1 at low, medium and high concentrations was 78.98% to 93.51%, 81.88% to 100.74%, and 82.19% to 116.12%, respectively, with RSDs ranging from 1.1% to 6.6%. For Polygalae Radix, a homogenizer was a better means to extract aflatoxins before enriching them with the immunoaffinity columns. Among the 28 batches of Polygalae Radix samples, 2 batches of crude drugs were found to be substandard. The highest total content of aflatoxins was 313.63 μg·kg-1 and the AFB1 content was as high as 143.27 μg·kg-1. Conclusion A simple and reliable new method for simultaneous determination of four aflatoxins in Polygalae Radix medicinal materials has been established, which can overcome the flaws of existing detection methods currently available and expose the risk of aflatoxin contamination in Polygalae Radix.

Key words: Polygalae Radix, Aflatoxins, High Performance Liquid Chromatography-Fluorescence Detector, Matrix Effects

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