DNA metabarcoding identification of prescription ingredients in traditional medicine Ruyi Jinhuang San

Shi, Linchun; Liu, JinXin; Wei, MiaoJie; Xie, Lifang; Song, Jingyuan

doi:10.1360/n052017-00204

Cited by 2 publications

(2 citation statements)

References 13 publications

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“…DNA metabarcoding is currently the most widely used detection method for mixed biological samples. Many studies are available that involve the identification of biological ingredients of traditional herbal medicine based on DNA metabarcoding technology ( Jia et al, 2017 ; Xin et al, 2018b ; Shi et al, 2018 ; Zhang et al, 2020 ). However, the PCR amplification efficiency of universal DNA barcode primers is affected by the severe DNA degradation of traditional herbal medicine ( Xin et al, 2018a ).…”

Section: Discussionmentioning

confidence: 99%

“…Adulterants and potential contaminant species were also found in the commercial samples, indicating that this method can effectively detect the biological components of Chinese patent medicines ( Xin et al, 2018b ). Furthermore, a study based on high-throughput sequencing and ITS2 regions detected some prescription ingredient adulterants (Cangzhu and Tiannanxing) in traditional Ruyi Jinhuang San medicine ( Shi et al, 2018 ). However, minimal studies are available involving the species identification in traditional herbal patent medicine based on shotgun sequencing.…”

Section: Introductionmentioning

confidence: 99%

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The Species Identification in Traditional Herbal Patent Medicine, Wuhu San, Based on Shotgun Metabarcoding

Liu

Shi

et al. 2021

Front. Pharmacol.

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Traditional herbal patent medicine typically consists of multiple ingredients, making it challenging to supervise contamination by impurities and the improper use of raw materials. This study employed shotgun metabarcoding for the species identification of biological ingredients in traditional herbal patent medicine, Wuhu San. The five prescribed herbal materials found in Wuhu San were collected, and their reference sequences were obtained by traditional DNA barcoding using Sanger sequencing. Two lab-made and three commercial Wuhu San samples were collected, and a total of 37.14 Gb of shotgun sequencing data was obtained for these five samples using the Illumina sequencing platform. A total of 1,421,013 paired-end reads were enriched for the Internal Transcribed Spacer 2 (ITS2), psbA and trnH intergenic spacer region (psbA-trnH), maturase k (matK), and ribulose-1, 5-bisphosphate carboxylase (rbcL) regions. Furthermore, 80, 11, 9, and 8 operational taxonomic units were obtained for the ITS2, psbA-trnH, matK, and rbcL regions, respectively, after metagenomic assembly, annotation, and chimeric detection. In the two lab-made mock samples, all labeled ingredients in the Wuhu San prescription were successfully detected, and the positive control, Panax quinquefolius L., was detected in the HSZY172 mock sample. Three species, namely Angelica sinensis (Oliv.) Diels, Saposhnikovia divaricata (Turcz. ex Ledeb.) Schischk., and Carthamus tinctorius L., belonging to three labeled ingredients, Angelicae Sinensis Radix (Danggui), Saposhnikoviae Radix (Fangfeng), and Carthami Flos (Honghua), were detected in the three commercial samples. Angelica dahurica (Hoffm.) Benth. & Hook. f. ex Franch. & Sav., the original Angelicae Dahuricae Radix (Baizhi) species, was only detected in WHS003. Arisaema erubescens (Wall.) Schott, Arisaema heterophyllum Blume, or Arisaema amurense Maxim., the original Arisaematis Rhizoma (Tiannanxing) species, were not detected in any of the commercial samples, which could be attributed to the fact that this medicinal material underwent extensive processing. In addition, the Saposhnikovia divaricata adulterant was detected in all the commercial samples, while 24 fungal genera, including Aspergillus, were identified in both the lab-made and commercial samples. This study showed that shotgun metabarcoding provided alternative strategy and technical means for identifying prescribed ingredients in traditional herbal patent medicine and displayed the potential to effectively complement traditional methods.

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