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Comparison of DNA barcoding and metabarcoding approaches for the authentication of herbal infusions in the Portuguese market

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Resumo:The authentication of commercial herbal products such as herbal teas and infusions is essential to ensure consumer safety and product integrity. In this study, 100 commercial herbal infusions, including 94 single-species products and six polyherbal formulations, were analysed using DNA barcoding (matK and rbcL) and ITS2 metabarcoding. Moreover, the performance of two custom reference databases (NCBI and BOLD) was compared in ITS2 metabarcoding. DNA extraction was successful for 94 samples, while six single-species products failed to amplify any of the tested barcodes. Among the 88 remaining single-species samples, ITS2 showed the highest amplification success (100 %), outperforming the barcodes rbcL (94 %) and matK (84 %). Sanger sequencing confirmed the labelled species in 69.3 % of cases with rbcL and 48.9 % with matK. While 63 samples would be considered authentic solely based on barcoding (i.e., if either rbcL or matK matched the label), ITS2 metabarcoding revealed that many of these contained additional undeclared species, indicating that barcoding alone overestimated product authenticity. Of the 85 samples successfully analysed by ITS2 metabarcoding, only 27 (32 %) fully matched their label, while 58 (68 %) contained either additional undeclared species or complete substitutions. Several products contained undeclared species in significant proportions, indicating potential economic adulteration. Analysis of mock-mixtures demonstrated that ITS2 metabarcoding can approximate ingredient proportions, though recovery bias may occur. Overall, the results revealed (i) the importance of curated and comprehensive databases, with a higher number of species being identified by NCBI database, (ii) the superior sensitivity of ITS2 metabarcoding, and (iii) the widespread mislabelling in commercial herbal products.
Autores principais:Pinto, M. Alice
Outros Autores:Santos, Joana; Quaresma, Andreia; Honrado, Mónica; Rufino, José; Amaral, Joana S.
Assunto:Authenticity Infusions ITS2 metabarcoding MatK RbcL
Ano:2026
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Instituto Politécnico de Bragança
Idioma:inglês
Origem:Biblioteca Digital do IPB
Descrição
Resumo:The authentication of commercial herbal products such as herbal teas and infusions is essential to ensure consumer safety and product integrity. In this study, 100 commercial herbal infusions, including 94 single-species products and six polyherbal formulations, were analysed using DNA barcoding (matK and rbcL) and ITS2 metabarcoding. Moreover, the performance of two custom reference databases (NCBI and BOLD) was compared in ITS2 metabarcoding. DNA extraction was successful for 94 samples, while six single-species products failed to amplify any of the tested barcodes. Among the 88 remaining single-species samples, ITS2 showed the highest amplification success (100 %), outperforming the barcodes rbcL (94 %) and matK (84 %). Sanger sequencing confirmed the labelled species in 69.3 % of cases with rbcL and 48.9 % with matK. While 63 samples would be considered authentic solely based on barcoding (i.e., if either rbcL or matK matched the label), ITS2 metabarcoding revealed that many of these contained additional undeclared species, indicating that barcoding alone overestimated product authenticity. Of the 85 samples successfully analysed by ITS2 metabarcoding, only 27 (32 %) fully matched their label, while 58 (68 %) contained either additional undeclared species or complete substitutions. Several products contained undeclared species in significant proportions, indicating potential economic adulteration. Analysis of mock-mixtures demonstrated that ITS2 metabarcoding can approximate ingredient proportions, though recovery bias may occur. Overall, the results revealed (i) the importance of curated and comprehensive databases, with a higher number of species being identified by NCBI database, (ii) the superior sensitivity of ITS2 metabarcoding, and (iii) the widespread mislabelling in commercial herbal products.