Authentication of Herbal Supplements Using Next-Generation Sequencing

Ivanova, Natalia V.; Kuzmina, Maria L.; Braukmann, Thomas; Borisenko, Alex; Zakharov, Evgeny

doi:10.1371/journal.pone.0156426

Cited by 100 publications

(149 citation statements)

References 113 publications

(110 reference statements)

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“…Next-Generation Sequencing (NGS) represents a new vision to detect the organic ingredients in mixed samples, such as tablets, powders, capsules and pills545556, and will provide an efficient and cost-effective mean to detect species composition of mixed TCM products. Targeting both DNA and characteristic components by DNA sequencing and chromatographic analysis will be a better integrative approach for authenticating TCM products, especially for standardized extracts with degraded DNA57.…”

Section: Resultsmentioning

confidence: 99%

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Shi

Zhao

Yao

et al. 2017

Sci Rep

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Pulsatillae radix is a conventional traditional Chinese medicine (TCM) with common name Baitouweng, and has notable effects on inflammation and dysentery. Pulsatilla chinensis (Bge.) Regel is the only source plant of Baitouweng recorded in Chinese Pharmacopoeia, but its adulteration often occurs in the market that possibly affects medicinal efficacy and safety. We have established an internal transcribed spacer 2 (ITS2) barcode library based on 105 plant samples from 12 Pulsatilla species and 10 common adulterants. Results indicate that ITS2 barcoding can accurately distinguish Pulsatilla species from their adulterants. Pulsatilla chinensis can be discriminated from 11 congeneric species by two stable single nucleotide polymorphisms (SNPs) in the ITS2 region. Additionally, a quick specific PCR-RFLP identification assay based on the ITS2 barcode was developed. Using specific primers ITS2/PR1 combined with restriction enzyme Bgl I, Pu. chinensis can rapidly be differentiated from other species via simple and low-cost test procedures. Furthermore, 30 commercial Baitouweng products were tested and only two products were derived from authentic Pu. chinensis. Thus, these two molecular approaches provide practical tools for quick identification of commercial Baitouweng products and can help ensure the safe use of this TCM product.

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Section: Resultsmentioning

confidence: 99%

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Shi

Zhao

Yao

et al. 2017

Sci Rep

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“…A more stringent threshold of 1% would cluster up to 5.3% of Habenaria species into single MOTUs, and only 0.8% of Disa and 1.4% of Satyrium species. Studies such as Ivanova et al (2016) that use a clustering threshold of 2%, risk overlooking species diversity among closely related species in their metabarcoding downstream analyses. Something that needs to be considered as well, however, is sequence inaccuracies due to PCR amplification and sequencing errors.…”

Section: Discussionmentioning

confidence: 99%

“…So far the only plant studies to use nrITS in metabarcoding have used nrITS2 to determine the taxonomic composition of pollen collected by honey bees (Galimberti et al 2014;Richardson et al 2015) and species composition in herbal medicines (Cheng et al 2014;Ivanova et al 2016).…”

Section: Introductionmentioning

confidence: 99%

High-throughput sequencing of African chikanda cake highlights conservation challenges in orchids

et al. 2017

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Chikanda is a traditional dish made with wild-harvested ground orchid tubers belonging to three orchidioid genera, Disa, Satyrium and Habenaria, all of which are CITES appendix II-listed. Identification of collected orchid tubers is very difficult and documentation of constituent species in prepared chikanda has hitherto been impossible. Here amplicon metabarcoding was used in samples of six prepared chikanda cakes to study genetic sequence diversity and species diversity in this product. Molecular operational taxonomic unit identification using similarity-matching reveals that species of all three genera were present in the chikanda samples studied. Disa was present in all of the samples, Satyrium in five out of six and Habenaria in one of the samples, as well as a number of other plants. The fact that each sample contained orchids and the presence of a wide variety of species from all genera in this traditional dish raise serious concerns about the sustainability of this trade and the future of wild orchid populations in the main harvest areas. This proof-of-concept study shows that Ion-Torrent PGM is a cost-effective scalable platform for metabarcoding using the relatively long nrITS1 and nrITS2 regions. 123Biodivers Conserv (2017) 26:2029-2046 DOI 10.1007/s10531-017-1343 Furthermore, nrITS metabarcoding can be successfully used for the detection of specific ingredients in a highly-processed food product at genus level, and this makes it a useful tool in the detection of possible conservation issues arising from commercialized trade or processed plant products.

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“…This is due to its inability to resolve mixed signals from such kind of samples [258]. However, such limitation can be overcome by involving the use of next-generation sequencing (NGS) which has several advantages over the Sanger sequencing.…”

Section: Dna Barcodingmentioning

confidence: 99%

Review: DNA Barcoding and Chromatography Fingerprints for the Authentication of Botanicals in Herbal Medicinal Products

Abubakar

Salleh²,

Shamsir³

et al. 2017

Evidence-Based Complementary and Alternative Medicine

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In the last two decades, there has been a tremendous increase in the global use of herbal medicinal products (HMPs) due to their claimed health benefits. This has led to increase in their demand and consequently, also, resulted in massive adulteration. This is due to the fact that most of the traditional methods cannot identify closely related species in a process product form. Therefore the urgent need for simple and rapid identification methods resulted in the discovery of a novel technique. DNA barcoding is a process that uses short DNA sequence from the standard genome for species identification. This technique is reliable and is not affected by external factors such as climates, age, or plant part. The difficulties in isolation of DNA of high quality in addition to other factors are among the challenges encountered using the DNA barcoding in the authentication of HMP. These limitations indicated that using DNA barcoding alone may ineffectively authenticate the HMP. Therefore, the combination of DNA barcoding with chromatographic fingerprint, a popular and generally accepted technique for the assessment and quality control of HMP, will offer an efficient solution to effectively evaluate the authenticity and quality consistency of HMP. Detailed and quality information about the main composition of the HMPs will help to ascertain their efficacy and safety as these are very important for quality control.

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Authentication of Herbal Supplements Using Next-Generation Sequencing

Cited by 100 publications

References 113 publications

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

Rapidly discriminate commercial medicinal Pulsatilla chinensis (Bge.) Regel from its adulterants using ITS2 barcoding and specific PCR-RFLP assay

High-throughput sequencing of African chikanda cake highlights conservation challenges in orchids

Review: DNA Barcoding and Chromatography Fingerprints for the Authentication of Botanicals in Herbal Medicinal Products

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