Preprint / Version 1

Biomonitoring for traditional herbal medicinal products using DNA metabarcoding and single molecule, real-time sequencing

Authors

  • Tianyi Xin aKey Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100193, China
  • Zhichao Xu aKey Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100193, China
  • Jing Jia aKey Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100193, China
  • Christine Leon bRoyal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK
  • Songnian Hu cBeijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China
  • Yulin Lin aKey Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100193, China
  • Subramanyam Ragupathy dNHP Research Alliance, Biodiversity Institute of Ontario (BIO), University of Guelph, Guelph N1G 2W1, Ontario, Canada
  • Jingyuan Song aKey Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100193, China
  • Steven Newmaster dNHP Research Alliance, Biodiversity Institute of Ontario (BIO), University of Guelph, Guelph N1G 2W1, Ontario, Canada

Keywords:

KEY WORDS: Traditional herbal medicinal products (THMP), Species mixture, Authentication, DNA metabarcoding, Single molecule real-time (SMRT) sequencing, Circular-consensus sequencing (CCS)

Abstract

Global concerns have been paid to the potential hazard of traditional herbal medicinal products (THMPs). Substandard and counterfeit THMPs, including traditional Chinese patent medicine, health foods, dietary supplements, etc. are potential threats to public health. Recent marketplace studies using DNA barcoding have determined that the current quality control methods are not sufficient for ensuring the presence of authentic herbal ingredients and detection of contaminants/adulterants. An efficient biomonitoring method for THMPs is of great needed. Herein, metabarcoding and single-molecule, real-time (SMRT) sequencing were used to detect the multiple ingredients in Jiuwei Qianghuo Wan (JWQHW), a classical herbal prescription widely used in China for the last 800 years. Reference experimental mixtures and commercial JWQHW products from the marketplace were used to confirm the method. Successful SMRT sequencing results recovered 5416 and 4342 circular-consensus sequencing (CCS) reads belonging to the ITS2 and psbA-trnH regions. The results suggest that with the combination of metabarcoding and SMRT sequencing, it is repeatable, reliable, and sensitive enough to detect species in the THMPs, and the error in SMRT sequencing did not affect the ability to identify multiple prescribed species and several adulterants/contaminants. It has the potential for becoming a valuable tool for the biomonitoring of multi-ingredient THMPs. KEY WORDS: Traditional herbal medicinal products (THMP), Species mixture, Authentication, DNA metabarcoding, Single molecule real-time (SMRT) sequencing, Circular-consensus sequencing (CCS)

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