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Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis

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...

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Main Authors: Fadzil, N. F., Wagiran, A., Salleh, F. M., Abdullah, S., Izham, N. H. M.
Format: Article
Language:English
Published: MDPI AG 2018
Subjects:
TP Chemical technology
Online Access:http://eprints.utm.my/id/eprint/79683/1/ShamsiahAbdullah2018_AuthenticityTestingandDetectionofEurycomalongifolia.pdf
http://eprints.utm.my/id/eprint/79683/
http://dx.doi.org/10.3390/genes9080408
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spelling my.utm.796832019-01-28T06:38:01Z http://eprints.utm.my/id/eprint/79683/ Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis Fadzil, N. F. Wagiran, A. Salleh, F. M. Abdullah, S. Izham, N. H. M. TP Chemical technology 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. MDPI AG 2018 Article PeerReviewed application/pdf en http://eprints.utm.my/id/eprint/79683/1/ShamsiahAbdullah2018_AuthenticityTestingandDetectionofEurycomalongifolia.pdf Fadzil, N. F. and Wagiran, A. and Salleh, F. M. and Abdullah, S. and Izham, N. H. M. (2018) Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis. Genes, 9 (8). ISSN 2073-4425 http://dx.doi.org/10.3390/genes9080408 DOI:10.3390/genes9080408
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
language English
topic TP Chemical technology
spellingShingle TP Chemical technology
Fadzil, N. F.
Wagiran, A.
Salleh, F. M.
Abdullah, S.
Izham, N. H. M.
Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
description 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.
format Article
author Fadzil, N. F.
Wagiran, A.
Salleh, F. M.
Abdullah, S.
Izham, N. H. M.
author_facet Fadzil, N. F.
Wagiran, A.
Salleh, F. M.
Abdullah, S.
Izham, N. H. M.
author_sort Fadzil, N. F.
title Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
title_short Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
title_full Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
title_fullStr Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
title_full_unstemmed Authenticity testing and detection of Eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
title_sort authenticity testing and detection of eurycoma longifolia in commercial herbal products using bar-high resolution melting analysis
publisher MDPI AG
publishDate 2018
url http://eprints.utm.my/id/eprint/79683/1/ShamsiahAbdullah2018_AuthenticityTestingandDetectionofEurycomalongifolia.pdf
http://eprints.utm.my/id/eprint/79683/
http://dx.doi.org/10.3390/genes9080408
_version_ 1643658262769303552
score 13.211869

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