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Development of a loop-mediated isothermal amplification assay for the rapid detection of Styphnolobium japonicum (L.) Schott as an adulterant of Ginkgo biloba (L.)

2024Journal article Open access
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dc.contributor.authorRodrigues, Vânia
dc.contributor.authorHonrado, Mónica
dc.contributor.authorSantos, Joana
dc.contributor.authorPinto, M. Alice
dc.contributor.authorAmaral, Joana S.
dc.contributor.authorRodrigues, Vânia
dc.date.accessioned2024-06-13T16:02:25Z
dc.date.available2024-06-13T16:02:25Z
dc.date.issued2024
dc.description.abstractSpecies adulteration is a concern in herbal products, especially when plant substitutes of lower economic value replace valuable botanicals. Styphnolobium japonicum is well known as a potential adulterant of Ginkgo biloba, which is one of the most demanded medicinal plants due to its wide use in pharmaceuticals, food supplements, and traditional medicine. Despite bearing some resemblance to ginkgo's flavonol composition, S. japonicum lacks many of G. biloba's desired therapeutic properties. To prevent adulteration practices, it is crucial to implement rigorous quality control measures, including fast and simple diagnostic tools that can be used on-field. Purpose: This study aims to develop for the first time a species-specific loop-mediated isothermal amplification (LAMP) method for the fast identification of S. japonicum in ginkgo-containing products. Methods: A set of four specific primers (SjF3, SjB3, SjFIP, and SjBIP) and loop primers (SjLF and SjLB) were designed for a LAMP based assay using the 5.8S partial sequence and the internal transcribed spacer 2 of nuclear ribosomal DNA of S. japonicum. Results: The successful amplification of the LAMP assay was inspected through visual detection, with the highest intensity recorded at the optimal conditions set at 68 °C for 40 min. The primers showed high specificity and were able to accurately discriminate S. japonicum from G. biloba and 49 other species of medicinal plants. Furthermore, the proposed LAMP assay proved to be fast, selective, and highly sensitive, as demonstrated by the absolute and relative limits of detection, which were reached at 0.5 pg for S. japonicum DNA and 0.01 % S. japonicum in G. biloba, respectively. Conclusions: This novel approach allows easy identification and discrimination of S. japonicum as a potential adulterant of G. biloba, thus being a useful tool for quality control. Compared to chromatographic or PCR-based methods, the assay proved to be fast, sensitive and did not require expensive equipment, thus offering the possibly usage in field analysis.pt_PT
dc.description.sponsorshipThe authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through the project “POIROT: novel methods and approaches for detecting the illegal addition of Pharmaceutical drugs and bOtanIcal adulteRatiOn in planT food supplements” (PTDC/SAU-PUB/3803/2021) and through national funds FCT/MCTES (PIDDAC) to CIMO (UIDB/00690/2020 and UIDP/ 00690/2020) and SusTEC (LA/P/0007/2020). M. Honrado is grateful to FCT for PhD grant 2021.08119.BD financed by POPH-QREN (subsidised by FSE and MCTES).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationRodrigues, Vânia; Honrado, Mónica; Santos, Joana; Pinto, M. Alice; Amaral, Joana S. (2024). Development of a loop-mediated isothermal amplification assay for the rapid detection of Styphnolobium japonicum (L.) Schott as an adulterant of Ginkgo biloba (L.). Phytomedicine. ISSN 0944-7113. 128, p. 1-10pt_PT
dc.identifier.doi10.1016/j.phymed.2023.155322pt_PT
dc.identifier.issn0944-7113
dc.identifier.urihttp://hdl.handle.net/10198/29904
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationMountain Research Center
dc.relationMountain Research Center
dc.relationAssociate Laboratory for Sustainability and Tecnology in Mountain Regions
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectAdulterationpt_PT
dc.subjectAuthenticitypt_PT
dc.subjectColorimetric detectionpt_PT
dc.subjectGinkgo bilobapt_PT
dc.subjectLAMPpt_PT
dc.subjectSophora japonicapt_PT
dc.titleDevelopment of a loop-mediated isothermal amplification assay for the rapid detection of Styphnolobium japonicum (L.) Schott as an adulterant of Ginkgo biloba (L.)pt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleMountain Research Center
oaire.awardTitleMountain Research Center
oaire.awardTitleAssociate Laboratory for Sustainability and Tecnology in Mountain Regions
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FSAU-PUB%2F3803%2F2021/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00690%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F00690%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0007%2F2020/PT
oaire.citation.endPage10pt_PT
oaire.citation.startPage1pt_PT
oaire.citation.titlePhytomedicinept_PT
oaire.citation.volume128pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
person.familyNameHonrado
person.familyNamePinto
person.familyNameAmaral
person.familyNameRodrigues
person.givenNameMónica
person.givenNameM. Alice
person.givenNameJoana S.
person.givenNameVânia
person.identifier.ciencia-id4712-B40B-4B0E
person.identifier.ciencia-idF814-A1D0-8318
person.identifier.ciencia-id5319-7DE8-BEDA
person.identifier.ciencia-id6B14-50D7-7EBC
person.identifier.orcid0000-0002-5126-4693
person.identifier.orcid0000-0001-9663-8399
person.identifier.orcid0000-0002-3648-7303
person.identifier.orcid0000-0003-2044-4277
person.identifier.scopus-author-id8085507800
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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