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DNA barcoding and NMR spectroscopy-based assessment of species adulteration in the raw herbal trade of Saraca asoca (Roxb.) Willd, an important medicinal plant

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Abstract

Saraca asoca (Roxb.) Willd, commonly known as “Asoka” or “Ashoka,” is one of the most important medicinal plants used in raw herbal trade in India. The bark extracts of the tree are used in the treatment of leucorrhea and other uterine disorders besides also having anti-inflammatory, anti-bacterial, anti-pyretic, anti-helminthic, and analgesic activity. The indiscriminate and rampant extraction of the wood to meet the ever-increasing market demand has led to a sharp decline in naturally occurring populations of the species in the country. Consequently, the species has recently been classified as “vulnerable” by the International Union for Conservation of Nature (IUCN). Increasing deforestation and increasing demand for this medicinal plant have resulted in a limited supply and suspected widespread adulteration of the species in the raw herbal trade market. Adulteration is a serious concern due to: (i) reduction in the efficacy of this traditional medicine, (ii) considerable health risk to consumers, and (iii) fraudulent product substitution that impacts the economy for the Natural Health Product (NHP) Industry and consumers. In this paper, we provide the first attempt to assess the extent of adulteration in the raw herbal trade of S. asoca using DNA barcoding validated by NMR spectroscopic techniques. Analyzing market samples drawn from 25 shops, mostly from peninsular India, we show that more than 80 % of the samples were spurious, representing plant material from at least 7 different families. This is the first comprehensive and large-scale study to demonstrate the widespread adulteration of market samples of S. asoca in India. These results pose grave implications for the use of raw herbal drugs, such as that of S. asoca, on consumer health and safety. Based on these findings, we argue for a strong and robust regulatory framework to be put in place, which would ensure the quality of raw herbal trade products and reassure consumer confidence in indigenous medicinal systems.

DNA barcoding and NMR spectroscopy-based assessment of adulteration in Saraca asoca.

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Acknowledgments

This work was supported by Department of Biotechnology, Government of India (Grant number: No.BT/IN/ ISTP-EOI/2011). The NMR experiments were performed on a 600-MHz Avance-III FT-NMR spectrometer at the NMR Research Facility, IISER Mohali. Saraca asoca samples from Odisha were kindly provided by Dr. Pratap Panda, RPRC, Bhubaneswar

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Authors and Affiliations

  1. School of Ecology and Conservation, University of Agricultural Sciences, GKVK, Bangalore, 560065, India

    Santhosh Kumar Jayanthinagar Urumarudappa & Uma Shaanker Ramanan

  2. Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore, 560065, India

    Santhosh Kumar Jayanthinagar Urumarudappa & Uma Shaanker Ramanan

  3. Department of Post Graduate Studies and Research in Biotechnology, Jnanasahyadri, Kuvempu University, Shankaraghatta, Shimoga, 577451, India

    Santhosh Kumar Jayanthinagar Urumarudappa & Krishna Venkatarangaiah

  4. Indian Institute of Science Education and Research (IISER) Mohali, Knowledge City Sector 81, PO Manauli, Mohali, 140306, Punjab, India

    Navdeep Gogna & Kavita Dorai

  5. Centre for Biodiversity Genomics (CBG), College of Biological Sciences, Department of Integrative Biology, University of Guelph, Toronto, N1G 2W1, Canada

    Steven G. Newmaster & Ragupathy Subramanyam

  6. Ashoka Trust for Research in Ecology and the Environment, Royal Enclave, Srirampura, Jakkur Post, Bangalore, 560064, India

    Seethapathy Gopalakrishnan Saroja, Ravikanth Gudasalamani & Uma Shaanker Ramanan

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  1. Santhosh Kumar Jayanthinagar Urumarudappa
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  2. Navdeep Gogna
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ESM 1

S1 Fig. 1 2D DOSY 1H NMR spectrum of (a) plant bark sample and (b) raw drug sample (HAS 318) showing specific NMR resonances for phenolics, carbohydrates, lipids and amino acid groups. Codes of the BRM and HAS are given in Table 1 and Table 2. S1 Fig. 2 (A) PLS-DA score plots (Component 1 vs Component 2) for comparing Saraca asoca bark samples with all drug samples. Codes of the BRM and HAS are given in Table 1 and Table 2. (DOC 182 kb)

ESM 2

S1 Fig. 3 OPLS-DA score plot for authentic plant sample and HAS 220 showing one predictive and one orthogonal component. (PDF 17 kb)

ESM 3

S1 Fig. 4 OPLS-DA score plot for authentic plant sample and HAS 476 showing one predictive and one orthogonal component. (PDF 18 kb)

ESM 4

S1 Fig. 5 NMR spectra for authentic plant sample, HAS 220 and HAS 476 in the aromatic region. Rectangular box and star (*) indicate regions of spectra similar in both authentic plant sample and HAS 220. (PDF 31 kb)

ESM 5

S1 Table 1 Query match of rbcL and psbA-trnH gene sequences of Saraca asoca trade samples in NCBI database. (XLSX 11 kb)

ESM 6

S1 Table 2 Metabolites present in 1 H NMR and 2D NMR spectra of Saraca asoca bark extract, with chemical shift given in ppm and the corresponding multiplicity and scalar coupling J values (in Hz). S1 Table 3 Post hoc analysis showing which drugs are different in group A, given the p value threshold of 0.05. S1 Table 4 Post hoc analysis showing which drugs are different in group B, given the p value threshold of 0.05. S1 Table 5 Metabolites contributing to differences between authentic plant samples (BRM) and market sample HAS 220. S1 Table 6: Metabolites contributing to differences between authentic plant samples (BRM) and market sample HAS 476. (DOC 87 kb)

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Urumarudappa, S.K.J., Gogna, N., Newmaster, S.G. et al. DNA barcoding and NMR spectroscopy-based assessment of species adulteration in the raw herbal trade of Saraca asoca (Roxb.) Willd, an important medicinal plant. Int J Legal Med 130, 1457–1470 (2016). https://doi.org/10.1007/s00414-016-1436-y

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