Comparative Study of the Effect of Sample Pretreatment and Extraction on the Determination of Flavonoids from Lemon (Citrus limon)
Identifieur interne : 000231 ( Pmc/Checkpoint ); précédent : 000230; suivant : 000232Comparative Study of the Effect of Sample Pretreatment and Extraction on the Determination of Flavonoids from Lemon (Citrus limon)
Auteurs : Carlos A. Ledesma-Escobar [Espagne] ; Feliciano Priego-Capote [Espagne] ; María D. Luque De Castro [Espagne]Source :
- PLoS ONE [ 1932-6203 ] ; 2016.
Abstract
Flavonoids have shown to exert multiple beneficial effects on human health, being also appreciated by both food and pharmaceutical industries. Citrus fruits are a key source of flavonoids, thus promoting studies to obtain them. Characteristics of these studies are the discrepancies among sample pretreatments and among extraction methods, and also the scant number of comparative studies developed so far.
Evaluate the effect of both the sample pretreatment and the extraction method on the profile of flavonoids isolated from lemon.
Extracts from fresh, lyophilized and air-dried samples obtained by shaking extraction (SE), ultrasound-assisted extraction (USAE), microwave-assisted extraction (MAE) and superheated liquid extraction (SHLE) were analyzed by LC–QTOF MS/MS, and 32 flavonoids were tentatively identified using MS/MS information. ANOVA applied to the data from fresh and dehydrated samples and from extraction by the different methods revealed that 26 and 32 flavonoids, respectively, were significant (
The present study shows clear discrimination caused by both sample pretreatments and extraction methods. Under the studied conditions, liophilization provides extracts with higher amounts of flavonoids, and USAE is the best method for isolation of these compounds, followed by MAE and SE. On the contrary, the SHLE method was the less favorable to extract flavonoids from citrus owing to degradation.
Url:
DOI: 10.1371/journal.pone.0148056
PubMed: 26807979
PubMed Central: 4726533
Affiliations:
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Ledesma-Escobar</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation></author><author><name sortKey="Priego Capote, Feliciano" sort="Priego Capote, Feliciano" uniqKey="Priego Capote F" first="Feliciano" last="Priego-Capote">Feliciano Priego-Capote</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation></author><author><name sortKey="Luque De Castro, Maria D" sort="Luque De Castro, Maria D" uniqKey="Luque De Castro M" first="María D." last="Luque De Castro">María D. Luque De Castro</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba, Spain</addr-line></nlm:aff><country xml:lang="fr">Espagne</country><wicri:regionArea>Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba</wicri:regionArea><wicri:noRegion>Córdoba</wicri:noRegion></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec id="sec001"><title>Background</title><p>Flavonoids have shown to exert multiple beneficial effects on human health, being also appreciated by both food and pharmaceutical industries. Citrus fruits are a key source of flavonoids, thus promoting studies to obtain them. Characteristics of these studies are the discrepancies among sample pretreatments and among extraction methods, and also the scant number of comparative studies developed so far.</p></sec><sec id="sec002"><title>Objective</title><p>Evaluate the effect of both the sample pretreatment and the extraction method on the profile of flavonoids isolated from lemon.</p></sec><sec id="sec003"><title>Results</title><p>Extracts from fresh, lyophilized and air-dried samples obtained by shaking extraction (SE), ultrasound-assisted extraction (USAE), microwave-assisted extraction (MAE) and superheated liquid extraction (SHLE) were analyzed by LC–QTOF MS/MS, and 32 flavonoids were tentatively identified using MS/MS information. ANOVA applied to the data from fresh and dehydrated samples and from extraction by the different methods revealed that 26 and 32 flavonoids, respectively, were significant (<italic>p</italic>≤0.01). The pairwise comparison (Tukey HSD; <italic>p</italic>≤0.01) showed that lyophilized samples are more different from fresh samples than from air-dried samples; also, principal component analysis (PCA) showed a clear discrimination among sample pretreatment strategies and suggested that such differences are mainly created by the abundance of major flavonoids. On the other hand, pairwise comparison of extraction methods revealed that USAE and MAE provided quite similar extracts, being SHLE extracts different from the other two. In this case, PCA showed a clear discrimination among extraction methods, and their position in the scores plot suggests a lower abundance of flavonoids in the extracts from SHLE. In the two PCA the loadings plots revealed a trend to forming groups according to flavonoid aglycones.</p></sec><sec id="sec004"><title>Conclusions</title><p>The present study shows clear discrimination caused by both sample pretreatments and extraction methods. Under the studied conditions, liophilization provides extracts with higher amounts of flavonoids, and USAE is the best method for isolation of these compounds, followed by MAE and SE. On the contrary, the SHLE method was the less favorable to extract flavonoids from citrus owing to degradation.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Tripoli, E" uniqKey="Tripoli E">E Tripoli</name></author><author><name sortKey="Guardia, Ml" uniqKey="Guardia M">ML Guardia</name></author><author><name sortKey="Giammanco, S" uniqKey="Giammanco S">S Giammanco</name></author><author><name sortKey="Majo, Dd" uniqKey="Majo D">DD Majo</name></author><author><name sortKey="Giammanco, M" uniqKey="Giammanco M">M Giammanco</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Davis, Wb" uniqKey="Davis W">WB Davis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Herrmann, K" uniqKey="Herrmann K">K Herrmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Roohbakhsh, A" uniqKey="Roohbakhsh A">A 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<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, CA USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26807979</article-id><article-id pub-id-type="pmc">4726533</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0148056</article-id><article-id pub-id-type="publisher-id">PONE-D-15-33944</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Agriculture</subject><subj-group><subject>Crop Science</subject><subj-group><subject>Crops</subject><subj-group><subject>Fruits</subject><subj-group><subject>Lemons</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Plants</subject><subj-group><subject>Fruits</subject><subj-group><subject>Lemons</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Agriculture</subject><subj-group><subject>Crop Science</subject><subj-group><subject>Crops</subject><subj-group><subject>Fruits</subject><subj-group><subject>Citrus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Plants</subject><subj-group><subject>Fruits</subject><subj-group><subject>Citrus</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Agriculture</subject><subj-group><subject>Crop Science</subject><subj-group><subject>Crops</subject><subj-group><subject>Fruits</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Plants</subject><subj-group><subject>Fruits</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Mathematical and Statistical Techniques</subject><subj-group><subject>Statistical Methods</subject><subj-group><subject>Multivariate Analysis</subject><subj-group><subject>Principal Component Analysis</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Physical Sciences</subject><subj-group><subject>Mathematics</subject><subj-group><subject>Statistics (Mathematics)</subject><subj-group><subject>Statistical Methods</subject><subj-group><subject>Multivariate Analysis</subject><subj-group><subject>Principal Component Analysis</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Physical Sciences</subject><subj-group><subject>Chemistry</subject><subj-group><subject>Chemical Compounds</subject><subj-group><subject>Phenols</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Physical Sciences</subject><subj-group><subject>Chemistry</subject><subj-group><subject>Chemical Reactions</subject><subj-group><subject>Ionization</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Mathematical and Statistical Techniques</subject><subj-group><subject>Statistical Methods</subject><subj-group><subject>Analysis of Variance</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Physical Sciences</subject><subj-group><subject>Mathematics</subject><subj-group><subject>Statistics (Mathematics)</subject><subj-group><subject>Statistical Methods</subject><subj-group><subject>Analysis of Variance</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Biochemistry</subject><subj-group><subject>Antioxidants</subject></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Comparative Study of the Effect of Sample Pretreatment and Extraction on the Determination of Flavonoids from Lemon (<italic>Citrus limon</italic>)</article-title><alt-title alt-title-type="running-head">Effect of Sample Pretreatment and Extraction on Flavonoids from Lemon</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Ledesma-Escobar</surname><given-names>Carlos A.</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="author-notes" rid="currentaff001"><sup>¤</sup></xref></contrib><contrib contrib-type="author"><name><surname>Priego-Capote</surname><given-names>Feliciano</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff003"><sup>3</sup></xref><xref ref-type="corresp" rid="cor001">*</xref></contrib><contrib contrib-type="author"><name><surname>Luque de Castro</surname><given-names>María D.</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff003"><sup>3</sup></xref><xref ref-type="corresp" rid="cor001">*</xref></contrib></contrib-group><aff id="aff001"><label>1</label><addr-line>Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, Córdoba, Spain</addr-line></aff><aff id="aff002"><label>2</label><addr-line>University of Córdoba Agroalimentary Excellence Campus, ceiA3, Campus of Rabanales, E-14071, Córdoba, Spain</addr-line></aff><aff id="aff003"><label>3</label><addr-line>Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba, Spain</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Abe</surname><given-names>Keiko</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>The University of Tokyo, JAPAN</addr-line></aff><author-notes><fn fn-type="conflict" id="coi001"><p><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p></fn><fn fn-type="con" id="contrib001"><p>Conceived and designed the experiments: CALE FPC MDLC. Performed the experiments: CALE. Analyzed the data: CALE FPC MDLC. Contributed reagents/materials/analysis tools: FPC MDLC. Wrote the paper: CALE FPC MDLC.</p></fn><fn fn-type="current-aff" id="currentaff001"><label>¤</label><p>Current address: Chemical and Biochemical Engineering Department, Technological Institute of Veracruz, Av. Miguel Angel de Quevedo 2779, Veracruz, Ver. 91897, México</p></fn><corresp id="cor001">* E-mail: <email>q72prcaf@uco.es</email> (FPC); <email>qa1lucam@uco.es</email> (MDLC)</corresp></author-notes><pub-date pub-type="epub"><day>25</day><month>1</month><year>2016</year></pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><volume>11</volume><issue>1</issue><elocation-id>e0148056</elocation-id><history><date date-type="received"><day>2</day><month>8</month><year>2015</year></date><date date-type="accepted"><day>27</day><month>11</month><year>2015</year></date></history><permissions><copyright-statement>© 2016 Ledesma-Escobar et al</copyright-statement><copyright-year>2016</copyright-year><copyright-holder>Ledesma-Escobar et al</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><self-uri content-type="pdf" xlink:type="simple" xlink:href="pone.0148056.pdf"></self-uri><abstract><sec id="sec001"><title>Background</title><p>Flavonoids have shown to exert multiple beneficial effects on human health, being also appreciated by both food and pharmaceutical industries. Citrus fruits are a key source of flavonoids, thus promoting studies to obtain them. Characteristics of these studies are the discrepancies among sample pretreatments and among extraction methods, and also the scant number of comparative studies developed so far.</p></sec><sec id="sec002"><title>Objective</title><p>Evaluate the effect of both the sample pretreatment and the extraction method on the profile of flavonoids isolated from lemon.</p></sec><sec id="sec003"><title>Results</title><p>Extracts from fresh, lyophilized and air-dried samples obtained by shaking extraction (SE), ultrasound-assisted extraction (USAE), microwave-assisted extraction (MAE) and superheated liquid extraction (SHLE) were analyzed by LC–QTOF MS/MS, and 32 flavonoids were tentatively identified using MS/MS information. ANOVA applied to the data from fresh and dehydrated samples and from extraction by the different methods revealed that 26 and 32 flavonoids, respectively, were significant (<italic>p</italic>≤0.01). The pairwise comparison (Tukey HSD; <italic>p</italic>≤0.01) showed that lyophilized samples are more different from fresh samples than from air-dried samples; also, principal component analysis (PCA) showed a clear discrimination among sample pretreatment strategies and suggested that such differences are mainly created by the abundance of major flavonoids. On the other hand, pairwise comparison of extraction methods revealed that USAE and MAE provided quite similar extracts, being SHLE extracts different from the other two. In this case, PCA showed a clear discrimination among extraction methods, and their position in the scores plot suggests a lower abundance of flavonoids in the extracts from SHLE. In the two PCA the loadings plots revealed a trend to forming groups according to flavonoid aglycones.</p></sec><sec id="sec004"><title>Conclusions</title><p>The present study shows clear discrimination caused by both sample pretreatments and extraction methods. Under the studied conditions, liophilization provides extracts with higher amounts of flavonoids, and USAE is the best method for isolation of these compounds, followed by MAE and SE. On the contrary, the SHLE method was the less favorable to extract flavonoids from citrus owing to degradation.</p></sec></abstract><funding-group><funding-statement>The Spanish Ministerio de Economía y Competitividad (MINECO) and FEDER program are thanked for financial support through Project CTQ2012-37428. FPC was sponsored by Ministerio de Ciencia e Innovación (MICINN) through a Ramón y Cajal Contract (RYC-2009-03921). CALE was sponsored by Mexican National Council for Science and Technology (CONACYT No. 224905, México).</funding-statement></funding-group><counts><fig-count count="4"></fig-count><table-count count="3"></table-count><page-count count="16"></page-count></counts><custom-meta-group><custom-meta id="data-availability"><meta-name>Data Availability</meta-name><meta-value>All relevant data are within the paper and its Supporting Information files.</meta-value></custom-meta></custom-meta-group></article-meta><notes><title>Data Availability</title><p>All relevant data are within the paper and its Supporting Information files.</p></notes></front></pmc><affiliations><list><country><li>Espagne</li></country></list><tree><country name="Espagne"><noRegion><name sortKey="Ledesma Escobar, Carlos A" sort="Ledesma Escobar, Carlos A" uniqKey="Ledesma Escobar C" first="Carlos A." last="Ledesma-Escobar">Carlos A. Ledesma-Escobar</name></noRegion><name sortKey="Ledesma Escobar, Carlos A" sort="Ledesma Escobar, Carlos A" uniqKey="Ledesma Escobar C" first="Carlos A." last="Ledesma-Escobar">Carlos A. Ledesma-Escobar</name><name sortKey="Luque De Castro, Maria D" sort="Luque De Castro, Maria D" uniqKey="Luque De Castro M" first="María D." last="Luque De Castro">María D. Luque De Castro</name><name sortKey="Luque De Castro, Maria D" sort="Luque De Castro, Maria D" uniqKey="Luque De Castro M" first="María D." last="Luque De Castro">María D. Luque De Castro</name><name sortKey="Luque De Castro, Maria D" sort="Luque De Castro, Maria D" uniqKey="Luque De Castro M" first="María D." last="Luque De Castro">María D. Luque De Castro</name><name sortKey="Priego Capote, Feliciano" sort="Priego Capote, Feliciano" uniqKey="Priego Capote F" first="Feliciano" last="Priego-Capote">Feliciano Priego-Capote</name><name sortKey="Priego Capote, Feliciano" sort="Priego Capote, Feliciano" uniqKey="Priego Capote F" first="Feliciano" last="Priego-Capote">Feliciano Priego-Capote</name><name sortKey="Priego Capote, Feliciano" sort="Priego Capote, Feliciano" uniqKey="Priego Capote F" first="Feliciano" last="Priego-Capote">Feliciano Priego-Capote</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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