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Genes 2018, 9(8), 408; https://doi.org/10.3390/genes9080408

Authenticity Testing and Detection of Eurycoma longifolia in Commercial Herbal Products Using Bar-High Resolution Melting Analysis

1
Department of Biotechnology & Medical Engineering, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, Johor Bahru 81310, Malaysia
2
Faculty of Plantation and Agrotechnology, Universiti Teknologi MARA (UiTM), Shah Alam 40450, Selangor, Malaysia
*
Author to whom correspondence should be addressed.
Received: 3 June 2018 / Revised: 30 July 2018 / Accepted: 7 August 2018 / Published: 12 August 2018
(This article belongs to the Special Issue DNA Barcoding and Metabarcoding of Complex Matrices)
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Abstract

The present study demonstrated High Resolution Melting (HRM) analysis combined with DNA barcode (Bar-HRM) as a fast and highly sensitive technique for detecting adulterants in Eurycoma longifolia commercial herbal products. Targeting the DNA barcoding of the chloroplastic region-ribulose biphosphate carboxylase large chain (rbcL) and the nuclear ribosomal region- internal transcribed spacer 2 (ITS2), PCR amplification and HRM analysis using saturated Eva green dye as the source of fluorescence signals, was accomplished by employing a real-time cycler. The results were further validated by sequencing to identify unknown sequence from Genbank database and to generate phylogenetic tree using neighbour joint (NJ) analysis. Both of the DNA markers exhibited a distinguishable melting temperature and shape of the normalised curve between the reference and the adulterants. In the case of species identification, ITS2 was more successful in differentiating between species. Additionally, detection of admixture sample containing small traces of targeted E. longifolia DNA (w/v) can be detected as low as 5% for rbcL and less than 1% for ITS2, proving the sensitivity and versatility of the HRM analysis. In conclusion, the Bar-HRM analysis is a fast and reliable technique that can effectively detect adulterants in herbal products. Therefore, this will be beneficial for regulatory agencies in order to regulate food safety issues. View Full-Text
Keywords: Eurycoma longifolia; rbcL; ITS2; authentication; adulteration; Bar-HRM Eurycoma longifolia; rbcL; ITS2; authentication; adulteration; Bar-HRM
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Fadzil, N.F.; Wagiran, A.; Mohd Salleh, F.; Abdullah, S.; Mohd Izham, N.H. Authenticity Testing and Detection of Eurycoma longifolia in Commercial Herbal Products Using Bar-High Resolution Melting Analysis. Genes 2018, 9, 408.

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