{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:42:11Z","timestamp":1760060531766,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T00:00:00Z","timestamp":1757030400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia and Minist\u00e9rio da Ci\u00eancia","doi-asserted-by":"publisher","award":["PTDC\/SAU-PUB\/3803\/2021","UIDB\/50006\/2020","CIMO UIDB\/00690\/2020","UIDP\/00690\/2020","LA\/P\/0007\/2020"],"award-info":[{"award-number":["PTDC\/SAU-PUB\/3803\/2021","UIDB\/50006\/2020","CIMO UIDB\/00690\/2020","UIDP\/00690\/2020","LA\/P\/0007\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Ginkgo biloba is one of the most consumed medicinal plants and broadly included as an ingredient in plant food supplements (PFS) and herbal infusions, being potential targets for economically motivated adulteration. This work aimed at comparing the use of DNA and phytochemical markers to authenticate the botanical origin of ginkgo-leaf extracts and PFS. Quantitative real-time PCR was used to detect ginkgo DNA, while ultra-high performance liquid chromatography with tandem mass spectrometry detection (UHPLC-MS\/MS) determined its main phytochemicals (terpene lactones and flavonol aglycones). DNA was detected in all ginkgo leaf extracts, mainly water, while the highest levels of phytochemicals were obtained using ethanol or acetone as solvents. The results suggested that 4 out of a total of 19 PFS samples were adulterated, with two samples evidencing the addition of quercetin from sources other than ginkgo. The other two samples showed low amounts of ginkgo phytochemicals, which was corroborated by low DNA content, suggesting the use of reduced amounts of G. biloba leaf material.<\/jats:p>","DOI":"10.3390\/foods14173111","type":"journal-article","created":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T14:53:01Z","timestamp":1757083981000},"page":"3111","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Towards Botanical Authentication of Ginkgo Food Supplements: A Holistic Approach Based on Phytochemical and Genomic Markers"],"prefix":"10.3390","volume":"14","author":[{"given":"Liliana","family":"Grazina","sequence":"first","affiliation":[{"name":"REQUIMTE\/LAQV, Faculdade de Farm\u00e1cia, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"}]},{"given":"Paula","family":"Pa\u00edga","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida 431, 4249-015 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3648-7303","authenticated-orcid":false,"given":"Joana S.","family":"Amaral","sequence":"additional","affiliation":[{"name":"CIMO, LA SusTEC, Instituto Polit\u00e9cnico de Bragan\u00e7a, Campus de Santa Apol\u00f3nia, 5300-253 Bragan\u00e7a, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8229-2902","authenticated-orcid":false,"given":"Joana","family":"Costa","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Faculdade de Farm\u00e1cia, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1500-9211","authenticated-orcid":false,"given":"Manuela M.","family":"Moreira","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida 431, 4249-015 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3924-776X","authenticated-orcid":false,"given":"Cristina","family":"Delerue-Matos","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida 431, 4249-015 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5311-8895","authenticated-orcid":false,"given":"Isabel","family":"Mafra","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Faculdade de Farm\u00e1cia, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.hermed.2016.04.003","article-title":"Adulteration and poor quality of Ginkgo biloba supplements","volume":"6","author":"Booker","year":"2016","journal-title":"J. Herb. Med."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Garcia-Alvarez, A., Egan, B., de Klein, S., Dima, L., Maggi, F.M., Isoniemi, M., Ribas-Barba, L., Raats, M.M., Meissner, E.M., and Badea, M. (2014). Usage of plant food supplements across six European countries: Findings from the PlantLIBRA consumer survey. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0092265"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1016\/j.fitote.2008.05.007","article-title":"Biology and chemistry of Ginkgo biloba","volume":"79","author":"Singh","year":"2008","journal-title":"Fitoterapia"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1016\/j.phymed.2014.01.010","article-title":"Adulteration of Ginkgo biloba products and a simple method to improve its detection","volume":"21","author":"Wohlmuth","year":"2014","journal-title":"Phytomedicine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.phymed.2018.04.009","article-title":"Anti-neuroinflammatory effects of Ginkgo biloba extract EGb761 in LPS-activated primary microglial cells","volume":"44","author":"Gargouri","year":"2018","journal-title":"Phytomedicine"},{"key":"ref_6","unstructured":"EMA (2025, January 15). European Medicines Agency\u2014Assessment Report on Ginkgo biloba L., Folium\u2014EMA\/HMPC\/321095\/2012-Amend. Available online: https:\/\/www.ema.europa.eu\/en\/documents\/herbal-report\/final-assessment-report-ginkgo-biloba-l-folium_en.pdf."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1055\/a-0581-5203","article-title":"Ginkgo biloba food supplements on the European market\u2014adulteration patterns revealed by quality control of selected samples","volume":"84","author":"Czigle","year":"2018","journal-title":"Planta Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1604","DOI":"10.1039\/D4NP00014E","article-title":"Estimating the extent of adulteration of the popular herbs black cohosh, echinacea, elder berry, ginkgo, and turmeric\u2014Its challenges and limitations","volume":"41","author":"Orhan","year":"2024","journal-title":"Nat. Prod. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.5740\/jaoacint.18-0319","article-title":"Validated identity test method for Ginkgo biloba NHPs using DNA-based species-specific hydrolysis PCR probe","volume":"102","author":"Dhivya","year":"2019","journal-title":"J. AOAC Int."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1139\/gen-2014-0130","article-title":"Authentication of Ginkgo biloba herbal dietary supplements using DNA barcoding","volume":"57","author":"Little","year":"2014","journal-title":"Genome"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Liu, Y., Wang, X.-Y., Wei, X.-M., Gao, Z.-T., and Han, J.-P. (2018). Rapid authentication of Ginkgo biloba herbal products using the recombinase polymerase amplification assay. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-26402-8"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Grazina, L., Amaral, J.S., Costa, J., and Mafra, I. (2020). Authentication of Ginkgo biloba herbal products by a novel quantitative real-time PCR approach. Foods, 9.","DOI":"10.3390\/foods9091233"},{"key":"ref_13","unstructured":"European Directorate for the Quality of Medicines (2019). European Pharmacopoeia, Council of Europe. [10th ed.]."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3870","DOI":"10.1111\/1541-4337.13222","article-title":"Mass spectrometry-based approaches to assess the botanical authenticity of dietary supplements","volume":"22","author":"Grazina","year":"2023","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1093\/jaoac\/92.2.645","article-title":"Column liquid chromatography\/electrospray ionization-time of flight-mass spectrometry and ultraperformance column liquid chromatography\/mass spectrometry methods for the determination of ginkgolides and bilobalide in the leaves of Ginkgo biloba and dietary supplements","volume":"92","author":"Avula","year":"2009","journal-title":"J. AOAC Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1631\/jzus.B1000179","article-title":"Molecular authentication of geo-authentic Scrophularia ningpoensis","volume":"12","author":"Chen","year":"2011","journal-title":"J. Zhejiang Univ. Sci. B"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1555","DOI":"10.1016\/j.phytochem.2008.01.026","article-title":"Fingerprint profile of Ginkgo biloba nutritional supplements by LC\/ESI-MS\/MS","volume":"69","author":"Ding","year":"2008","journal-title":"Phytochemistry"},{"key":"ref_18","first-page":"394","article-title":"Adulteration determining of pharmaceutical forms of Ginkgo biloba extracts from different international manufacturers","volume":"8","author":"Demirezer","year":"2014","journal-title":"Rec. Nat. Prod."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.jpba.2014.04.027","article-title":"Direct analysis of 18 flavonol glycosides, aglycones and terpene trilactones in Ginkgo biloba tablets by matrix solid phase dispersion coupled with ultra-high performance liquid chromatography tandem triple quadrupole mass spectrometry","volume":"97","author":"Liu","year":"2014","journal-title":"J. Pharmaceut. Biomed. Anal."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1016\/j.jpba.2019.03.058","article-title":"A sensitive and selective multiple reaction monitoring mass spectrometry method for simultaneous quantification of flavonol glycoside, terpene lactones, and biflavonoids in Ginkgo biloba leaves","volume":"170","author":"Wang","year":"2019","journal-title":"J. Pharmaceut. Biomed. Anal."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7733","DOI":"10.1007\/s00216-015-8938-1","article-title":"Identification of Ginkgo biloba supplements adulteration using high performance thin layer chromatography and ultra high performance liquid chromatography-diode array detector-quadrupole time of flight-mass spectrometry","volume":"407","author":"Avula","year":"2015","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/j.phymed.2016.02.003","article-title":"An effective identification and quantification method for Ginkgo biloba flavonol glycosides with targeted evaluation of adulterated products","volume":"23","author":"Ma","year":"2016","journal-title":"Phytomedicine"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6969","DOI":"10.1007\/s00216-020-02830-2","article-title":"A nontargeted approach to determine the authenticity of Ginkgo biloba L. plant materials and dried leaf extracts by liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and chemometrics","volume":"412","author":"Cruz","year":"2020","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1798","DOI":"10.1093\/jaoac\/93.6.1798","article-title":"Fingerprint analysis of Ginkgo biloba leaves and related health foods by high-performance liquid chromatography\/electrospray ionization-mass spectrometry","volume":"93","author":"Song","year":"2010","journal-title":"J. AOAC Int."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1016\/j.mcp.2015.06.002","article-title":"DNA extraction from plant food supplements: Influence of different pharmaceutical excipients","volume":"29","author":"Costa","year":"2015","journal-title":"Mol. Cell. Probe"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1016\/j.foodchem.2017.02.136","article-title":"Novel quantitative real-time PCR approach to determine safflower (Carthamus tinctorius) adulteration in saffron (Crocus sativus)","volume":"229","author":"Villa","year":"2017","journal-title":"Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.jpba.2016.08.032","article-title":"Blood-brain barrier specific permeability assay reveals N-methylated tyramine derivatives in standardised leaf extracts and herbal products of Ginkgo biloba","volume":"131","author":"Rendes","year":"2016","journal-title":"J. Pharmaceut. Biomed. Anal."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"111875","DOI":"10.1016\/j.foodres.2022.111875","article-title":"Comparative study on the phenolic composition and in vitro bioactivity of medicinal and aromatic plants from the Lamiaceae family","volume":"161","author":"Caleja","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.ejps.2016.12.024","article-title":"Analysis of pharmaceutical adulterants in plant food supplements by UHPLC-MS\/MS","volume":"99","author":"Rodrigues","year":"2017","journal-title":"Eur. J. Pharmac. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.jpba.2016.12.013","article-title":"Development of a multi-residue method for the determination of human and veterinary pharmaceuticals and some of their metabolites in aqueous environmental matrices by SPE-UHPLC\u2013MS\/MS","volume":"135","author":"Santos","year":"2017","journal-title":"J. Pharmaceut. Biomed. Anal."},{"key":"ref_31","unstructured":"Wenzl, T., Haedrich, J., Schaechtele, A., Robouch, P., and Stroka, J. (2025, March 01). Guidance Document on the Estimation of LOD and LOQ for Measurements in the Field of Contaminants in Feed and Food. Available online: https:\/\/food.ec.europa.eu\/system\/files\/2017-05\/animal-feed-guidance_document_lod_en.pdf."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Kuli\u0107, \u017d., Lehner, M.D., and Dietz, G.P.H. (2022). Ginkgo biloba leaf extract EGb 761\u00ae as a paragon of the product by process concept. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.1007746"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_34","unstructured":"ENGL (2025, March 15). Definition of Minimum Performance Requirements for Analytical Methods of GMO Testing. European Network of GMO Laboratories, Join Research Centre, EURL, Available online: https:\/\/gmo-crl.jrc.ec.europa.eu\/doc\/MPR%20Report%20Application%2020_10_2015.pdf."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1002\/rcm.8024","article-title":"Improving negative liquid chromatography\/electrospray ionization mass spectrometry lipidomic analysis of human plasma using acetic acid as a mobile-phase additive","volume":"32","author":"Monnin","year":"2018","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1021\/ac0351670","article-title":"Favorable effects of weak acids on negative-ion electrospray ionization mass spectrometry","volume":"76","author":"Wu","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_37","first-page":"6","article-title":"Review of extraction techniques extraction methods: Microwave, ultrasonic, pressurized fluid, soxhlet extraction, etc","volume":"6","author":"Patel","year":"2019","journal-title":"Int. J. Adv. Res. Chem. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/S0021-9673(02)00172-3","article-title":"Chemical analysis of Ginkgo biloba leaves and extracts","volume":"967","year":"2002","journal-title":"J. Chromatogr. A"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2002","DOI":"10.1016\/j.chroma.2009.01.013","article-title":"Chemical analysis and quality control of Ginkgo biloba leaves, extracts, and phytopharmaceuticals","volume":"1216","author":"Montoro","year":"2009","journal-title":"J. Chromatogr. A"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1002\/nano.202100152","article-title":"DNA precipitation revisited: A quantitative analysis","volume":"3","author":"He","year":"2022","journal-title":"Nano Sel."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Andersen, O.M., and Markham, K.R. (2005). Flavonoids: Chemistry, Biochemistry and Applications, Taylor & Francis Group CRC Press. [1st ed.].","DOI":"10.1201\/9781420039443"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0031-9422(97)00450-0","article-title":"Distribution of ginkgolides and terpenoid biosynthetic activity in Ginkgo biloba","volume":"48","author":"Carrier","year":"1998","journal-title":"Phytochemistry"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"872","DOI":"10.3389\/fpls.2017.00872","article-title":"Combining metabolic profiling and gene expression analysis to reveal the biosynthesis site and transport of ginkgolides in Ginkgo biloba L","volume":"8","author":"Lu","year":"2017","journal-title":"Front. Plant Sci."}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/17\/3111\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:40:58Z","timestamp":1760035258000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/17\/3111"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,5]]},"references-count":43,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["foods14173111"],"URL":"https:\/\/doi.org\/10.3390\/foods14173111","relation":{},"ISSN":["2304-8158"],"issn-type":[{"type":"electronic","value":"2304-8158"}],"subject":[],"published":{"date-parts":[[2025,9,5]]}}}