Detection of Adulteration by Wedelia calendulacea in Eclipta alba through ISSR and RAPD Markers

Abstract

In the crude herbal drug market, the highly medicinal Eclipta alba, popularly known as “Bhringaraja”, is often adulterated with Wedelia calendulacea. To evaluate the ability of simple genetic fingerprinting methods to differentiate between the two species, Inter-Simple Sequence Repeat and Random Amplified Polymorphic DNA assays were performed on 30 individuals of E. alba and 4 individuals of W. calendulacea. Samples of W. calendulacea were found to cluster in clades that distinctly separated from E. alba and at least one band was identified for each of the two species that can serve as a marker for identification. Despite wide variations and a limited number of samples, phylogenetic analysis helped in differentiating the original from the adulterant plant. The study underscores the importance of simple DNA-based fingerprinting as a cost-effective method for the detection of adulteration in plants, particularly with respect to E. alba.

Key words

Eclipta alba - Wedelia calendulacea - Asteraceae - phylogenetics - adulteration - molecular markers

^* These authors contributed equally to this work.

• References

• 1 Nadkarni KM. Indian Materia Medica, Vol. 1. Bombay: Popular Prakashan; 1976: 469-470
• 2 Sarin YK. Illustrated Herbs of Ayurveda. New Delhi, India: Council of Scientific & Industrial Research (CSIR) and Indian council of Medical Research (ICMR); 1996: 263
• 3 Pithayanukul P, Laovachirasuwan S, Bavovada R, Pakmanee N, Suttisri R. Anti-venom potential of butanolic extract of Eclipta prostrata against Malayan pit viper venom. J Ethnopharmacol 2004; 90: 347-352
• 4 Khare CP. Indian medicinal Plants: an illustrated Dictionary. New York: Springer; 2007. 230. 716
• 5 Chunekar KC. Bhavaprakasha Nighantu-Hindi Vyakhya. Varanasi, India: Chaukhambha Bharati Academy; 2004: 429
• 6 Wagner H, Fessler B. [In vitro 5-lipoxygenase inhibition by Eclipta alba extracts and the coumestan derivative wedelolactone]. Planta Med 1986; 5: 374-377
• 7 Nehybova T, Smarda J, Daniel L, Brezovsky J, Benes P. Wedelolactone induces growth of breast cancer cells by stimulation of estrogen receptor signalling. J Steroid Biochem Mol Biol 2015; 152: 76-83
• 8 Dixit VK. Controversial Ayurvedic herbs. J Adv Pharm Technol Res 2011; 2: 78-80
• 9 Pandit N. Raja Nighantu. Ed: Tripati I. Varanasi, India: Chaukhamba Khrishnadas Academy; 1983
• 10 Bapalal V. Some controversial drugs in Indian medicine. Varanasi, India: Chaukambha Orientalia; 1982: 155-156
• 11 Lucas SK. Dravyaguna Vijñāna, Vol 2. Varanasi, India: Chaukambha Vishwabharati; 2008: 234
• 12 Govindaraghavan S, Hennell JR, Sucher NJ. From classical taxonomy to genome and metabolome: towards comprehensive quality standards for medicinal herb raw materials and extracts. Fitoterapia 2010; 83: 979-988
• 13 Newmaster SG, Grguric M, Shanmughanandhan D, Ramalingam S, Ragupathy S. DNA barcoding detects contamination and substitution in North American herbal products. BMC Med 2013; 11: 222
• 14 Seethapathy GS, Ganesh D, Santhosh Kumar JU, Senthilkumar U, Newmaster SG, Ragupathy S, Uma Shaanker R, Ravikanth G. 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 2014; 129: 693-700
• 15 Xin T, Li X, Yao H, Lin Y, Ma X, Cheng R, Song J, Ni L, Fan C, Chen S. Survey of commercial Rhodiola products revealed species diversity and potential safety issues. Sci Rep 2015; 5: 8337
• 16 Williamson EM, Chan K, Xu Q, Nachtergael A, Bunel V, Zhang L, Ouedraogo M, Nortier J, Qu F, Shaw D, Liu X, Stévigny C, Kahumba J, Pelkonen O, Duez P. Evaluating the safety of herbal medicines: Integrated toxicological approaches. Science 2015; 347: S47-S49
• 17 Quinto CA, Tinoco R, Hellberg RS. DNA barcoding reveals mislabeling of game meat species on the U.S. commercial market. Food Control 2016; 59: 386-392
• 18 Coutinho Moraes DF, Still DW, Lum MR, Hirsch AM. DNA-based authentication of botanicals and plant-derived dietary supplements: where have we been and where are we going?. Planta Med 2015; 81: 687-695
• 19 Mishra P, Kumar A, Nagireddy A, Mani DN, Shukla AK, Tiwari R, Sundaresan V. DNA barcoding: an efficient tool to overcome authentication challenges in the herbal market. Plant Biotechnol J 2015; 14: 8-21
• 20 Kiran U, Khan S, Mirza KJ, Ram M, Abdin MZ. SCAR markers: a potential tool for authentication of herbal drugs. Fitoterapia 2010; 81: 969-976
• 21 Osathanunkul M, Madesis P, de Boer H. Bar-HRM for authentication of plant-based medicines: evaluation of three medicinal products derived from Acanthaceae species. PLoS One 2015; 10: e0128476
• 22 Farrar JS, Wittwer CT. Extreme PCR: efficient and specific DNA amplification in 15–60 seconds. Clin Chem 2014; 61: 145-153
• 23 Tamhankar S, Ghate V, Raut A, Rajput B. Molecular profiling of “Chirayat” complex using Inter Simple Sequence Repeat (ISSR) markers. Planta Med 2009; 75: 1266-1270