DNA barcoding in authentication of herbal raw materials, extracts and dietary supplements: a perspective

  • D. B. Anantha Narayana
  • Sudhakar T. Johnson


Cultivation, supply, and availability of unadulterated plant raw material in botanical or herbal industry are a challenging task. Plant species that are either in short supply or in high demand are prone to adulteration and substitution. Such unscrupulous activity results in availability of substandard products with declining quality standards ultimately posing safety concerns and health of consumers. Raw materials which are in the form of dried, fragmented or powdered forms are major victims of adulteration. We present here DNA-based marker approach to authenticate botanicals. Recent research advances such as 2D-electrophoresis, microarray, next generation sequencing, high-resolution melting analysis, and PCR-based methods have been reviewed and their role in authentication of raw material, botanicals in herbal extracts, finished formulations, and ingredients in dietary supplements has been presented. Due to extraction processes such as steaming, extraction at extreme conditions and presence of secondary metabolites original DNA integrity are not preserved which lead to interference with DNA extraction and PCR amplification, thus making DNA unsuitable for further molecular analysis. There is a scope to develop and validate DNA mini-barcodes for finished formulations. DNA barcode along with chromatographic methods, metabolomics, proteomics, and genomics has to be used to resolve substitution and quantity of raw material in ambiguity. Emerging regulatory scenario on DNA barcode in various Pharmacopoeia is presented. Though several advanced methods of botanical authentication are available, an industry always expects simple, cost efficient, and quick method to resolve authentication issues.


DNA barcode Botanicals authentication Herbal extracts Herbal raw material Regulatory status 



TSJ thanks Sri Koneru Raja Harin, Vice President, K. L. Education Foundation and Head, Department of Biotechnology, K. L. E. F University for the constant support. The authors have consulted several industry leaders and leading researchers in the field of DNA barcode whose inputs were valuable. We thank them for their support. The authors acknowledge Prof. K. Satyamoorthy and Kalpalatha Rai of Manipal School of Life Sciences, Manipal (Karnataka, India) for undertaking DNA barcode testing and providing the results. We thank Prof. Ambujakshi Reddy of Acharya and B N Reddy College of Pharmacy, Bangalore for providing the photographs of the samples of Nirgundi and permission to use them in this review.

Compliance with ethical standards

Conflict of interest

Authors declare that there is no conflict of interest.


  1. Agarwal A, D’Souza P, Johnson TS, Dethe SM, Chandrasekaran CV (2014) Use of in vitro assays for assessing botanicals. Curr Op Biotechnol 25:39–44CrossRefGoogle Scholar
  2. Bosmali I, Ganopoulos I, Madesis P, Tsaftaris A (2012) Microsatellite and DNA-barcode regions typing combined with high resolution melting (HRM) analysis for food forensic uses: a case study on lentils (Lens culinaris). Food Res Int 46:141–147CrossRefGoogle Scholar
  3. British Pharmacopoeia (2017a) Deoxyribonucleic acid (DNA) based identification techniques for herbal drugs. British Pharmacopoeia Appendix XI V, pp. V-A 354–357Google Scholar
  4. British Pharmacopoeia (2017b) British Pharmacopoeia Appendix V-A, Supplementary Chapter VII-D, pp. V-A 840–843Google Scholar
  5. Bruni I, De Mattia F, Galimberti A, Galasso G, Banfi E, Casiraghi M, Labra M (2010) Identification of poisonous plants by DNA barcoding approach. Int J Legal Med 124:595–603CrossRefGoogle Scholar
  6. Busconi M, Foroni C, Corradi M, Bongiorni C, Cattapan F, Fogher C (2003) DNA extraction from olive oil and its use in the identification of the production cultivar. Food Chem 83:127–134CrossRefGoogle Scholar
  7. CBOL Plant Working Group (2009) A DNA barcode for land plants. Proc Natl Acad Sci 106:12794–12797CrossRefGoogle Scholar
  8. Chen S, Yao H, Han J, Liu C, Song J, Shi L, Zhu Y, Ma X, Gao T, Pang X, Luo K, Li Y, Li X, Jia X, Lin Y, Leon C (2010) Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One 5:e8613CrossRefGoogle Scholar
  9. Chinese Pharmacopoeia Commission (2017) Guidelines for Molecular DNA Barcoding of Chinese Materia Medica. Pharmacopoeia of the People’s Republic of China Vol. IV. China Medical Science Press, pp. 411–413Google Scholar
  10. Cimino M (2010) Successful isolation and PCR amplification of DNA from National Institute of Standards and Technology herbal dietary supplement standard reference material powders and extracts. Planta Med 76:495–497CrossRefGoogle Scholar
  11. Costa J, Campos B, Amaral JS, Nunes ME, Oliveira MBPP, Mafra I (2016) HRM analysis targeting ITS1 and matK loci as potential DNA mini-barcodes for the authentication of Hypericum perforatum and Hypericum androsaemum in herbal infusions. Food Control 61:105–114CrossRefGoogle Scholar
  12. Gafner S, Blumenthal M, Reynaud D, Foster S, Techen N (2013) ABC review and critique of the research article—DNA barcoding detects contamination and substitution in North American herbal products—by Newmaster et al. Herbal-E-Gram, 10:11Google Scholar
  13. Gao T, Yao H, Song J, Liu C, Zhu Y, Ma X, Pang X, Xu H, Chen S (2010a) Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2. J Ethnopharmacol 130:116–121CrossRefGoogle Scholar
  14. Gao T, Yao H, Song J, Zhu Y, Liu C, Chen S (2010b) Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the large Asteraceae family. BMC Evol Biol 10:324CrossRefGoogle Scholar
  15. Gao T, Sun Z, Yao H, Song J, Zhu Y, Ma X, Chen S (2011) Identification of Fabaceae plants using the DNA barcode matK. Planta Med 77:92–94CrossRefGoogle Scholar
  16. Gao Z, Liu Y, Wang X, Song J, Chen S, Ragupathy S, Han J, Newmaster SG (2017) Derivative technology of DNA barcoding (nucleotide signature and SNP double peak methods) detects adulterants and substitution in Chinese patent medicines. Sci Rep 7:5858CrossRefGoogle Scholar
  17. Gonzalez MA, Baraloto C, Engel J, Mori SA, Petronelli P, Riera B, Roger A, Thebaud C, Chaveet J (2009) Identification of Amazonian trees with DNA barcodes. PLoS ONE 4:e7483CrossRefGoogle Scholar
  18. Gryson N, Ronsse F, Messens K, De Loose M, Verleyen T, Dewettinck K (2002) Detection of DNA during the refining of soybean oil. J Am Oil Chem Soc 79:171–174CrossRefGoogle Scholar
  19. Guo X, Wang X, Su W, Zhang G, Zhou R (2011) DNA barcodes for discriminating the medicinal plant Scutellaria baicalensis (Lamiaceae) and its adulterants. Biol Pharm Bull 34:1198–1203CrossRefGoogle Scholar
  20. Hamilton JP, Buell CR (2012) Advances in plant genome sequencing. Plant J 70:177–190CrossRefGoogle Scholar
  21. Hebert PDN, Cywinska A, Ball SL, de Waard JR (2003) Biological identifications through DNA barcodes. Proc Biol Sci 270:313–321CrossRefGoogle Scholar
  22. Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W (2004) Ten species in one: DNA barcoding reveals cryptic species in neotropical skipper butterfly Astraptes fulgerator. Proc Natl Acad Sci USA 101:14812–14817CrossRefGoogle Scholar
  23. Heinrich M, Anagnostou S (2017) From pharmacognosia to DNA-based medicinal plant authentication pharmacognosy through the centuries. Planta Med 83:1110–1116CrossRefGoogle Scholar
  24. Hellebrand M, Nagy M, Mörsel JT (1998) Determination of DNA traces in rapeseed oil. Eur Food Res Technol 206:237–242Google Scholar
  25. Heubl G (2010) New aspects of DNA based authentication of Chinese medicinal plants by molecular biology techniques. Planta Med 76:1963–1974CrossRefGoogle Scholar
  26. Ivanova NV, Kuzmina ML, Braukmann TWA, Borisenko AV, Zakharov EV (2016) Authentication of herbal supplements using next generation sequencing. PLoS ONE 11:e0156426CrossRefGoogle Scholar
  27. Jia J, Xu Z, Xin T, Shi L, Song J (2017) Quality control of the traditional patent medicine Yimu Wan based on SMRT sequencing and DNA barcoding. Front Plant Sci 8:926CrossRefGoogle Scholar
  28. Jiang C, Cao L, Yuan Y, Chen M, Jin Y, Huang L (2014) Barcoding melting curve analysis for rapid, sensitive, and discriminating authentication of Saffron (Crocus sativus L.) from its adulterants. Bio Med Res Int 1:809037Google Scholar
  29. Johnson TS, Agarwal RK, Agarwal A (2013) Non-timber forest products as a source of livelihood option for forest dwellers role of society, herbal industries and government agencies. Curr Sci 104:440–443Google Scholar
  30. Kang J, Lee S, Kang S, Kwon HN, Park JH, Kwon SW, Park S (2008) NMR- based metabolomics approach for the differentiation of Ginseng (Panax ginseng) roots from different origins. Arch Pharm Res 31:330–336CrossRefGoogle Scholar
  31. Kerr KCR, Stoeckle MY, Dove CJ, Weigt LA, Francis CM, Hebert PDN (2007) Comprehensive DNA barcode coverage of North American birds. Mol Ecol Notes 7:535–543CrossRefGoogle Scholar
  32. Khan S, Mirza KJ, Tayaab MD, Abdin MZ (2009) RAPD profile for authentication of medicinal plant Glycyrrhiza glabra Linn. Med Arom Plant Sci Biotechnol 3:48–51Google Scholar
  33. Kress JW, Wurdack KJ, Zimmer EA, Weigt LA, Janzen DH (2005) Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci 102:8369–8374CrossRefGoogle Scholar
  34. Kumar SJU, Krishna V, Seethapathy GS, Senthilkumar U, Ragupathy S, Ganeshaiah K, Ganesan KN, Newmaster SG, Ravikanth G, Uma Shaanker R (2015) DNA barcoding to assess species adulteration in raw drug trade of “Bala” (genus: Sida L.) herbal products in South India. Biochem Syst Ecol 61:501–509CrossRefGoogle Scholar
  35. Li DZ, Gao LM, Li HT, Wang H, Ge XJ, Liu JQ, Chen ZD et al (2011) Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants. Proc Natl Acad Sci USA 108:19641–19646CrossRefGoogle Scholar
  36. Lissy KP (2004) A comprehensive investigation into some selected species of Sida Linn, with reference to their substitutes and adulterants. Ph. D thesis. Mahatma Gandhi University, Kottayam, India (Kerala)Google Scholar
  37. Little DP (2014) Authentication of Ginkgo biloba herbal dietary supplements using DNA barcoding. Genome 57:513–516CrossRefGoogle Scholar
  38. Little DP, Jeanson ML (2013) DNA barcode authentication of saw palmetto herbal dietary supplements. Sci Rep 3:3518CrossRefGoogle Scholar
  39. Liu J, Shi L, Han J, Li G, Lu H, Hou J, Zhou X, Meng F, Downie SR (2014) Identification of species in the angiosperm family Apiaceae using DNA barcodes. Mol Ecol Resour 14:1231–1238CrossRefGoogle Scholar
  40. Liu Y, Wang X, Wang L, Chen X, Pang X, Han J (2016) A nucleotide signature for the identification of American ginseng and its products. Front Plant Sci 7:319Google Scholar
  41. Lum JHK, Fung KL, Cheung PY, Wong MS, Lee CH, Kwok FSL (2002) Proteome of oriental ginseng Panax ginseng C.A. Meyer and the potential to use it as an identification tool. Proteomics 2:1123–3038CrossRefGoogle Scholar
  42. Madesis P, Ganopoulos I, Ralli P, Tsaftaris A (2012) Barcoding the major Mediterranean leguminous crops by combining universal chloroplast and nuclear DNA sequence targets. Genet Mol Res 11:2548–2558CrossRefGoogle Scholar
  43. Narayana DBA, Johnson TS (2017) Emerging scenario on industrial use of bio resources and development of models for computing access and benefit sharing (ABS) mechanisms. J Biodivers Biopros Dev 4(2):165CrossRefGoogle Scholar
  44. Newmaster SG, Fazekas A, Ragupathy S (2006) DNA barcoding in the land plants: evaluation of rbcL in a multigene tiered approach. Can J Bot 84:335–341CrossRefGoogle Scholar
  45. Newmaster SG, Grguric M, Shanmughanandhan D, Ramalingam S, Ragupathy S (2013) DNA barcoding detects contamination and substitution in North American herbal products. BMC Med 11:222–234CrossRefGoogle Scholar
  46. Ogden R, McGough HN, Cowan RS, Chua L, Groves M, Mc Ewing R (2008) SNP based method for the genetic identification of ramin Gonystylus spp. timber and products: applied research meeting CITES enforcement needs. Endanger Species Res 9:255–261CrossRefGoogle Scholar
  47. Osathanunkul M, Suwannapoom C, Osathanunkul K, Madesise P, de Boer H (2016) Evaluation of DNA barcoding coupled high resolution melting for discrimination of closely related species in phytopharmaceuticals. Phytomedicine 23:156–165CrossRefGoogle Scholar
  48. Parvathy VA, Swetha VP, Sheeja TE, Leela NK, Chempakam B, Sasikumar B (2014) DNA barcoding to detect chilli adulteration in traded black pepper powder. Food Biotechnol 28:25–40CrossRefGoogle Scholar
  49. Rai PS, Bellampalli R, Dobriyal RM, Agarwal A, Satyamoorthy K, Narayana DBA (2012) DNA barcoding of authentic and substitute samples of herb of the family Asparagaceae and Asclepiadaceae based on the ITS2 region. J Ayurveda Integr Med 3:136–140CrossRefGoogle Scholar
  50. Reynaud DTH, Mishler BD, Neal-Kababick J, Brown PN (2015) The capabilities and limitations of DNA barcoding of botanical dietary supplements. Available at: Accessed 15 May 2015
  51. Santhosh Kumar JU, Gogna N, Newmaster SG, Venkatarangaiah K, Ragupathy S, Seethapathy GS (2016) 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 130:1457–1470CrossRefGoogle Scholar
  52. Seethapathy GS, Ganesh D, Santhosh Kumar JU, Senthilkumar U, Newmaster SG, Ragupathy S, Uma-Shaanker R, Ravikanth G (2015) Assessing product adulteration in natural health products for laxative yielding plants, Cassia, Senna, and Chamaecrista, in Southern India using DNA barcoding. Int J Legal Med 129:693–700CrossRefGoogle Scholar
  53. Smith MA, Poyarkov NA, Hebert PDN (2008) CO1 DNA barcoding amphibians: take the chance, meet the challenge. Mol Ecol Resour 8:235–246CrossRefGoogle Scholar
  54. Song J, Yao H, Li Y, Li X, Lin Y, Liu C, Han J, Xie C, Chen S (2009) Authentication of the family Polygonaceae in Chinese pharmacopoeia by DNA barcoding technique. J Ethnopharmacol 124:434–439CrossRefGoogle Scholar
  55. Srirama R, Senthilkumar U, Sreejayan N, Ravikanth G, Gurumurthy BR, Shivanna MB, Sanjappa M, Ganeshaiah KN, Uma Shaanker R (2010) Assessing species admixtures in raw drug trade of Phyllanthus, a hepato-protective plant using molecular tools. J Ethnopharmacol 130:208–215CrossRefGoogle Scholar
  56. Stoeckle MY, Gamble CC, Kirpekar R, Young G, Ahmed S, Little DP (2011) Commercial teas highlight plant DNA barcode identification successes and obstacles. Sci Rep 1:1–7CrossRefGoogle Scholar
  57. Sui XY, Huang Y, Tan Y, Guo Y, Long CL (2011) Molecular authentication of the ethnomedicinal plant Sabia parviflora and its adulterants by DNA barcoding technique. Planta Med 77:492–496CrossRefGoogle Scholar
  58. Sun Z, Gao T, Yao H, Shi L, Zhu Y, Chen S (2011) Identification of Lonicera japonica and its related species using the DNA barcoding method. Planta Med 77:301–306CrossRefGoogle Scholar
  59. Theodoridis S, Stefanaki A, Tezcan M, Aki C, Kokkini S, Vlachonasios KE (2012) DNA barcoding in native plants of the Labiatae (Lamiaceae) family from Chios Island (Greece) and the adjacent Cesme Karaburun Peninsula (Turkey). Mol Ecol Resour 12:620–633CrossRefGoogle Scholar
  60. Torelli A, Marieschi M, Bruni R (2014) Authentication of saffron (Crocus sativus L.) in different processed, retail products by means of SCAR markers. Food Control 36:126–131CrossRefGoogle Scholar
  61. USP (2018a) United States Pharmacopeial Convention (2018) United States Pharmacopeia, vol 4, 41st edn. United States Pharmacopeial Convention, Rockville, pp 6293–6304Google Scholar
  62. USP (2018b) United States Pharmacopoeia Convention (2018), United States Pharmacopeia, vol 5, 41st edn, Appendix 1. United States Pharmacopeial Convention, Rockville, pp 7357–7378Google Scholar
  63. Valentini A, Miquel C, Taberlet P (2010) DNA barcoding for honey biodiversity. Diversity 2:610–617CrossRefGoogle Scholar
  64. Vijayan K, Tsou CH (2010) DNA barcoding in plants: taxonomy in a new perspective. Curr Sci 99:1530–1541Google Scholar
  65. Wallace L, Boilard S, Eagle SHC, Spall JL, Shokralla S, Hajibabaei M (2012) DNA barcodes for everyday life: routine authentication of natural health products. Food Res Int 49:446–452CrossRefGoogle Scholar
  66. Wang CZ, Li P, Ding JY, Jin GQ, Yuan CS (2005) Identification of Fritillaria pallidiflora using diagnostic PCR and PCR–RFLP based on nuclear ribosomal DNA internal transcribed spacer sequences. Planta Med 1:384–386CrossRefGoogle Scholar
  67. Wang W, Wu Y, Yan Y, Ermakova M, Kerstetter R, Messing J (2010) DNA barcoding of the Lemnaceae, a family of aquatic monocots. BMC Plant Biol 10:205–216CrossRefGoogle Scholar
  68. Wang NA, Jacques FMB, Milne RI, Zhang CQ, Yang JB (2012) DNA barcoding of Nyssaceae (Cornales) and taxonomic issues. Bot Stud 53:265–274Google Scholar
  69. Wang X, Liu Y, Wang L, Han J, Chen S (2016) A nucleotide signature for the identification of Angelicae sinensis Radix (Danggui) and its products. Sci Rep 6:34940CrossRefGoogle Scholar
  70. Wittwer CT (2009) High-resolution DNA melting analysis: advancements and limitations. Hum Mutat 30:857–859CrossRefGoogle Scholar
  71. Xin T, Su C, Lin Y, Wang S, Xu Z, Song J (2018a) Precise species detection of traditional Chinese patent medicine by shotgun metagenomic sequencing. Phytomedicine 47:40–47CrossRefGoogle Scholar
  72. Xin T, Xu Z, Jia J, Leon C, Hu S, Lin Y, Ragupathy S, Song J, Newmaster SG (2018b) Biomonitoring for traditional herbal medicinal products using DNA metabarcoding and single molecule, real-time sequencing. Acta Pharm Sin B. 8:488–497CrossRefGoogle Scholar
  73. Yadav A, Javed A, Chaudhary AA, Ahmad A (2012) Development of Sequence Characterized Amplified Region (SCAR) marker for the authentication of Bacopa monnieri (L.) Wettst. Eur J Med Plants 2:186–198CrossRefGoogle Scholar
  74. Yang HQ, Dong YR, Gu ZJ, Liang N, Yang JB (2012) A preliminary assessment of matK, rbcL and trnH-psbA as DNA barcodes for Calamus (Arecaceae) species in China with a note on ITS. Ann Bot Fenn 49:319–330CrossRefGoogle Scholar
  75. Yu N, Gu H, Wei Y, Zhu N, Wang Y, Zhang H, Zhu Y, Zhang X, Ma C, Sun A (2016) Suitable DNA barcoding for identification and supervision of Piper kadsura in Chinese medicine markets. Molecules 21:1221CrossRefGoogle Scholar
  76. Zheng X, Cai D, Yao L, Teng Y (2008) Non-concerted ITS evolution, early origin and phylogenetic utility of ITS pseudogenes in Pyrus. Mol Phylogenet and Evol 48:892–903CrossRefGoogle Scholar
  77. Zuo Y, Chen Z, Kondo K, Funamoto T, Wen J, Zhou S (2011) DNA barcoding of Panax species. Planta Med 77:182–187CrossRefGoogle Scholar

Copyright information

© Korean Society for Plant Biotechnology 2019

Authors and Affiliations

  1. 1.Phytopharmaceuticals Group of Indian Pharmacopoeia Commission, Non Specified Food and Food Ingredients of Food Safety and Standards Authority of India (FSSAI)New DelhiIndia
  2. 2.Department of BiotechnologyCenter for Innovation, Incubation & Entrepreneurship, K. L. E. F. Deemed to be UniversityGuntur Dt.India
  3. 3.Coordinator, DST, NIDHI-Technology Business Incubator, K. L. E. F Deemed to be UniversityGuntur Dt.India
  4. 4.BangaloreIndia

Personalised recommendations