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Exploiting DNA markers for the authentication of Hypericum food supplements

Abstract

During the last years. the consumption of plant food supplements CPFS) containing medicinal plants has been growing in popularity. Consequently, there has been an increasing demand for plant material that can result in a higher number of frauds (substitution of a higher cost medicinal plant for a closely related. but cheaper species) and possibility of unintentional swap/misidentiflcation of plants. In botll cases. the PFS integrity. efficacy and safety are compromised. Therefore. methodologies for the unequivocal identification of plant species in PFS are required. In this work. DNA-markers were used to specifically identify Hypericum perforatum (used in for its antidepressive properties) and H. androsaemum (used as cholagogue and hepatic protector) in several PFS samples (tablets. capsules. tintures and ampoules). Different DNA extraction protocols. including in-house methods and commercial kits were tested. Tile extracts were amplified by real-time PCR targeting reference genes (universal eukaryotic and plant rubisco genes) and using species-specific primers targeting a DNA barcode loci CmatK gene). Best results were achieved for capsules an tablets using Nucleospin Plant 11 extraction method. whi le for liquid samples using an in-11ouse method based on DNA precipitation with ethanol and centrifugation. Although labeled. three samples tested negative for H. perfuratum. For some samples. negative amplification was obtained regard less of the targeted gene and DNA extraction method. pointing to some matnx interference. possibly due to DNA adsorption phenomena to pharmaceutica l excipients.This work was supported by projects EXPL/DTP-SAP/1438/2013 (4SaferPFS) and Pest-C/EQB/LA0006/2013 financed by FCT (FEDER funds through COMPETE). T J .R Fernandes and J Costa are grateful to FCT grants SFRH/BD/93711/2013 and SFRH/BPD/102404/2014info:eu-repo/semantics/publishedVersio

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