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1.河南中医药大学 药学院,郑州 450046
2.中国中医科学院 中药研究所,北京 100700
3.中国医学科学院 药用植物研究所,北京 100193
Received:20 February 2025,
Revised:2025-09-07,
Accepted:28 September 2025,
Online First:21 October 2025,
Published:20 May 2026
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张鸣宇,姜汶君,纪宝玉等.基于Herb-Q方法的当归药材及其加工品的分子定量检测[J].中国实验方剂学杂志,2026,32(10):192-200.
ZHANG Mingyu,JIANG Wenjun,JI Baoyu,et al.Quantitative Molecular Detection of Angelicae Sinensis Radix and Its Processed Products Based on Herb-Q Method[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(10):192-200.
张鸣宇,姜汶君,纪宝玉等.基于Herb-Q方法的当归药材及其加工品的分子定量检测[J].中国实验方剂学杂志,2026,32(10):192-200. DOI: 10.13422/j.cnki.syfjx.20251415.
ZHANG Mingyu,JIANG Wenjun,JI Baoyu,et al.Quantitative Molecular Detection of Angelicae Sinensis Radix and Its Processed Products Based on Herb-Q Method[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(10):192-200. DOI: 10.13422/j.cnki.syfjx.20251415.
目的
2
当归作为常用大宗药食同源药材,市场掺伪现象频发,严重影响制剂临床疗效。目前其混伪品定性鉴别技术已较为成熟,但其加工品掺伪定量检测方法尚属空白。该研究以当归及其主要近缘易混伪品“土当归”(朝鲜当归)为例开展定量检测研究,旨在建立当归及其加工品的草药分子定量检测方法(Herb-Q),为当归药材及其相关健康产品的定量检测技术的建立提供示范。
方法
2
基于叶绿体基因组全序列筛选当归及朝鲜当归的特异性单核苷酸多态性(SNP)位点。选取当归特异性位点通过不同掺伪比例的混合药材粉末进行定量方法学的考察(线性、定量限、检测限和重现性)。采用3个不同掺伪比例(15%、25%、35%)的活血止痛散验证Herb-Q方法对于含当归的中成药定量检测准确度。
结果
2
通过比对当归属123条叶绿体基因组序列,基于种内保守、种间特异及满足焦磷酸高通量测序要求的原则,确定叶绿体基因组序列NC_042826.1中的第9 674位点(A/G)和叶绿体基因组序列NC_029393.1中第38 592位点(T/C)可分别作为当归和朝鲜当归的专属性分子鉴别位点。选取当归专属性第9 674位点(A/G)进行所建立的Herb-Q方法线性关系
R
2
值为0.997 4(
R
2
>
0.99),线性关系良好。定量限与检测限均为2.0%,重现性较好[相对标准偏差(RSD)
<
2.0%]。基于Herb-Q方法构建的定量体系检测活血止痛散中伪品朝鲜当归掺入量,3次分子定量重复平均偏差为1.3%。
结论
2
该研究表明基于当归叶绿体基因组序列NC_042826.1中的第9 674位点(A/G)所构建的Herb-Q定量检测方法在当归和“土当归”及其在加工品中具有良好的适用性、客观性和准确性,可为市售当归衍生产品如中药制剂、膳食营养补充剂及保健品的定量检测提供技术支撑。
Objective
2
Angelicae Sinensis Radix, a commonly used medicinal herb with both medicinal and edible properties, is frequently adulterated in the market, severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature, quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant, "Tu Danggui" (
Angelica gigas
), was conducted to establish a herbal quantitative molecular detection (Herb-Q) method for Angelicae Sinensis Radix and its processed products, providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products.
Methods
2
The specific single-nucleotide polymorphism (SNP) loci of Angelicae Sinensis Radix
and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations (linearity, limit of quantification, limit of detection, and reproducibility) by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios (15%, 25%, and 35%) was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix.
Results
2
By comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix, based on the principles of intraspecies conservation, interspecies specificity, and meeting the requirements of pyrophosphate high-throughput sequencing, it was determined that 9 674
th
locus (A/G) in the chloroplast genome sequence NC_042826.1 and 38 592
nd
locus (T/C) in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular iden
tification loci of Angelicae Sinensis Radix
and Angelica gigas Nakai, respectively. The linear relationship
R
2
of the Herb-Q method established by selecting the specific 9 674
th
locus (A/G) of Angelicae Sinensis Radix was 0.997 4 (
R
2
>
0.99), indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%, exhibiting excellent reproducibility [relative standard deviation(RSD)
<
2.0%]. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit
A. gigas
in the Huoxue Zhitong powder, with an average deviation of 1.3% for three molecular quantitative replicates.
Conclusion
2
This research demonstrates that the Herb-Q quantitative detection method established based on the 9 674
th
locus (A/G) in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability, objectivity, and accuracy for Angelicae Sinensis Radix and
A. gigas
, and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives, including traditional Chinese medicinal preparations, dietary supplements, and nutraceuticals.
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