安全监测

利用多重连接探针扩增结合熔解曲线法鉴别人参与西洋参中掺杂的研究*

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  • 1.湖北省药品监督检验研究院,国家药品监督管理局中药质量控制重点实验室,武汉 430075;
    2.湖北中医药大学,武汉 430065;
    3.湖北大学生命科学学院,武汉 430062;
    4.武汉大学生命科学学院,武汉 430072
第一作者 Tel:18649345243;E-mail:1585919031@qq.com
**汪 波 Tel:13618638636;E-mail:wangbo1986@hotmail.com
杜 巍 Tel:13807190894;E-mail:wdu@whu.edu.cn

修回日期: 2022-04-17

  网络出版日期: 2024-06-24

基金资助

*湖北省青年拔尖人才培养计划;湖北省科技重大专项 (2020ACA007)“ 湖北中药质控标准物质研制及技术平台建立 ”;湖北省自然科学基金重点类项目“基于Q-marker的湖北特色道地药材质控关键技术研究”(2020CFA024);中国食品药品检定研究院项目“常见与重要中药材及饮片DNA分子鉴定研究项目(TCM2021-YJ11)”

Identification of mutual adulteration of Panax ginseng and Panax quinquefolius by multiplex ligation-dependent probe amplification combined with melting curve method*

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  • 1. Hubei Institute for Drug Control, NMPA Key Laboratory for Quality Control of Chinese Medicine, Wuhan 430075, China;
    2. Hubei University of Chinese Medicine, Wuhan 430060, China;
    3. School of Life Sciences, Hubei University, Wuhan 430062, China;
    4. College of Life Sciences, Wuhan University, Wuhan 430072, China

Revised date: 2022-04-17

  Online published: 2024-06-24

摘要

目的: 针对人参和西洋参饮片中相互掺杂问题建立快速检测鉴别方法。方法: 依据多重连接探针扩增(multiplex ligation-dependent probe amplification,MLPA)技术检测序列单核苷酸多态性(SNP)和点突变的原理,针对人参与西洋参的18S序列寻找SNP位点;设计特异性探针进行MLPA扩增,并由熔解曲线形成的熔解曲线峰得出熔解温度(melting temperature,Tm),根据Tm值的差异来鉴别人参与西洋参,并进行特异性、灵敏度、掺伪比实验,验证方法的可行性。结果: 人参探针的特异性熔解曲线Tm值为(79.3±0.2)℃,西洋参探针的熔解曲线Tm值为(81.2±0.2)℃。当西洋参探针与人参探针比例为0.6∶1.0时,人参掺杂10%的西洋参,或者西洋参中掺杂10%的人参均可被有效检出。结论: 本研究建立了稳定的人参与西洋参掺伪鉴别方法,该方法具有特异性强,灵敏度高的特点。

本文引用格式

王文斌, 莫静, 胡志刚, 杜巍, 汪波 . 利用多重连接探针扩增结合熔解曲线法鉴别人参与西洋参中掺杂的研究*[J]. 药物分析杂志, 2022 , 42(9) : 1577 -1585 . DOI: 10.16155/j.0254-1793.2022.09.11

Abstract

Objective: To establish a rapid detection and identification method for mutual adulteration in prepared slices of Panax ginseng and Panax quinquefolius. Methods: Based on the principle of using multiplex ligation-dependent probe amplification (MLPA) to detect single nucleotide polymorphism (SNP) and point mutation in sequences, a specific probe was designed to find SNP sites in the 18S sequences of Panax ginseng and Panax quinquefolius for MLPA amplification. The melting temperature (Tm) was derived from the peak of the melting curve, and the differences in Tm values were used to identify Panax ginseng and Panax quinquefolius. Specificity, sensitivity and adulteration ratio experiments were performed to verify the feasibility of the method. Results: The specific melting curve Tm value of Panax ginseng probe was (79.3±0.2)℃, and that of Panax quinquefolius probe was (81.2±0.2)℃. When the ratio of Panax quinquefolius probe to Panax ginseng probe was 0.6∶1.0.Panax ginseng adulterated with 10% Panax quinquefolius, or Panax quinquefolius adulterated with 10% ginseng could be effectively detected. Conclusion: This study establishesa stable method for the identification of Panax ginseng adulterated with Panax quinquefolius, which is specific and sensitive.

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