Department of Science and Technology, Gov. of Gujarat, Gujarat Biotechnology Research Centre (GBRC), Gandhinagar, India.
Phytochem Anal. 2021 Sep;32(5):804-810. doi: 10.1002/pca.3026. Epub 2021 Feb 1.
Ayurveda takes advantage of the beneficial properties of medicinal plants. High demands in combination with inadequate availability of botanicals and a lack of knowledge with respect to their precise identification lead to adulterations in herbal products. Identification becomes more difficult in complex herbal formulations. Four different polyherbal formulations have been analyzed for the present paper. The targeted plants have different pharmacological properties for various ailments.
We aimed to examine the rbcL gene based plant DNA mini-barcode to identify target and non-target plants in polyherbal formulations by using high-throughput next generation sequencing.
Degenerate primers of the selected mini-barcode region have been identified from the literature. A blend of 30 authentic medicinal plant species was used to examine the species resolution capacity of the mini-barcode. DNA was isolated from herbal formulations, an amplicon library was prepared, and sequencing was performed on an IonS5 system. Data were analyzed using various bioinformatics tools.
Analysis of control pooled samples revealed the optimum resolving power of the DNA mini-barcode. Data analysis of the commercial samples revealed that only one herbal formulation contained all plants and matched with listed contents. In two formulations, only 10 out of 21 and 11 out of 20 plants were detected, respectively. Additionally, several non-listed plants were also detected in these formulations. Two formulations contained >20% reads assigned to non-target plants. Overall, 21.98% of the reads were assigned to non-target plants.
The present study clearly demonstrated the successful application and potential of meta-barcoding in the quality control of complex herbal matrices. The results strongly suggest that this approach can be used in pharmacovigilance of processed herbal products.
阿育吠陀利用药用植物的有益特性。高需求与植物原料的供应不足以及对其准确鉴定的缺乏知识相结合，导致草药产品掺假。在复杂的草药配方中，鉴定变得更加困难。本文分析了四种不同的复方草药配方。靶向植物对各种疾病具有不同的药理特性。
我们旨在通过高通量下一代测序，利用 rbcL 基因的植物 DNA 微型条码来鉴定复方制剂中的目标和非目标植物。
从文献中确定了所选微型条码区域的简并引物。使用 30 种纯正药用植物混合物来检验微型条码的物种分辨率能力。从草药配方中提取 DNA，制备扩增子文库，并在 IonS5 系统上进行测序。使用各种生物信息学工具分析数据。
分析对照混合样本揭示了 DNA 微型条码的最佳分辨率能力。对商业样本的数据分析表明，只有一种草药配方包含所有植物，并与列出的成分相匹配。在两种配方中，分别仅检测到 21 种植物中的 10 种和 20 种植物中的 11 种。此外，这些配方中还检测到了几种未列出的植物。两种配方中超过 20%的读数被分配给非目标植物。总体而言，21.98%的读数被分配给非目标植物。
本研究清楚地证明了元条码在复杂草药基质质量控制中的成功应用和潜力。结果强烈表明，这种方法可用于加工草药产品的药物警戒。
