Exploring DNA quantity and quality from raw materials to botanical extracts
Subramanyam Ragupathy,
Adam C. Faller,
Dhivya Shanmughanandhan,
Prasad Kesanakurti,
R. Uma Shaanker,
Gudasalamani Ravikanth,
Ramalingam Sathishkumar,
Narayanasamy Mathivanan,
Jingyuan Song,
Jianping Han,
Steven Newmaster
Affiliations
Subramanyam Ragupathy
NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada; Corresponding author.
Adam C. Faller
NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
Dhivya Shanmughanandhan
NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
Prasad Kesanakurti
NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
R. Uma Shaanker
Department of Crop Physiology and School of Ecology and Conservation, University of Agricultural Sciences, GKVK, Bangalore, 560065, India
Gudasalamani Ravikanth
Conservation Genetics Lab, Ashoka Trust for Research in Ecology and the Environment (ATREE), Royal Enclave, Srirampura, Jakkur PO, Bengaluru, 560064, India
Ramalingam Sathishkumar
Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, 641046, India
Narayanasamy Mathivanan
Centre for Advanced Studies in Botany, University of Madras Guindy Campus, Chennai, 600 025, Tamil Nadu, India
Jingyuan Song
Engineering Research Center of Traditional Chinese Medicine Resource, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100193, China
Jianping Han
Engineering Research Center of Traditional Chinese Medicine Resource, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100193, China
Steven Newmaster
NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 2W1, Canada; Corresponding author.
Objectives: The aim of this study was to explore the variability in DNA quality and quantity along a gradient of industrial processing of botanical ingredients from raw materials to extracts. Methods: A data matrix was assembled for 1242 botanical ingredient samples along a gradient of industrial processing commonly used in the Natural Health Product (NHP) industry. Multivariate statistics was used to explore dependant variables for quality and quantity. The success of attaining a positive DNA test result along a gradient of industrial processing was compared among four biotechnologies: DNA barcoding, NGS, Sanger sequencing and qPCR. Results: There was considerable variance in DNA quality and quantity among the samples, which could be interpreted along a gradient from raw materials with greater quantities (50–120 ng/μL) of DNA and longer DNA (400-500bp) sequences to extracts, which were characterized by lower quantities (0.1–10.0 ng/μL) and short fragments (50-150bp). Conclusions: Targeted molecular diagnostic tests for species identity can be used in the NHP industry for raw and processed samples. Non-targeted tests or the use of NGS for any identity test needs considerable research and development and must be validated before it can be used in commercial operations as these methods are subject to considerable risk of false negative and positive results. Proper use of these tools can be used to ensure ingredient authenticity, and to avert adulteration, and contamination with plants that are a health concern. Lastly these tools can be used to prevent the exploitation of rare herbal species and the harvesting of native biodiversity for commercial purposes.
