Hydrophilic Interaction Liquid Chromatography Coupled to Mass Spectrometry and Multivariate Analysis of the De Novo Pyrimidine Pathway Metabolites.
Identifieur interne : 000479 ( Main/Exploration ); précédent : 000478; suivant : 000480Hydrophilic Interaction Liquid Chromatography Coupled to Mass Spectrometry and Multivariate Analysis of the De Novo Pyrimidine Pathway Metabolites.
Auteurs : Paula Galeano Garcia [Colombie] ; Barbara H. Zimmermann [Colombie] ; Chiara Carazzone [Colombie]Source :
- Biomolecules [ 2218-273X ] ; 2019.
Descripteurs français
- KwdFr :
- Analyse multifactorielle (MeSH), Chromatographie en phase liquide (MeSH), Concentration en ions d'hydrogène (MeSH), Interactions hydrophobes et hydrophiles (MeSH), Lycopersicon esculentum (microbiologie), Lycopersicon esculentum (métabolisme), Phytophthora infestans (physiologie), Pyrimidines (métabolisme), Spectrométrie de masse (MeSH), Température (MeSH).
- MESH :
- microbiologie : Lycopersicon esculentum.
- métabolisme : Lycopersicon esculentum, Pyrimidines.
- physiologie : Phytophthora infestans.
- Analyse multifactorielle, Chromatographie en phase liquide, Concentration en ions d'hydrogène, Interactions hydrophobes et hydrophiles, Spectrométrie de masse, Température.
English descriptors
- KwdEn :
- Chromatography, Liquid (MeSH), Hydrogen-Ion Concentration (MeSH), Hydrophobic and Hydrophilic Interactions (MeSH), Lycopersicon esculentum (metabolism), Lycopersicon esculentum (microbiology), Mass Spectrometry (MeSH), Multivariate Analysis (MeSH), Phytophthora infestans (physiology), Pyrimidines (metabolism), Temperature (MeSH).
- MESH :
- chemical , metabolism : Pyrimidines.
- metabolism : Lycopersicon esculentum.
- microbiology : Lycopersicon esculentum.
- physiology : Phytophthora infestans.
- Chromatography, Liquid, Hydrogen-Ion Concentration, Hydrophobic and Hydrophilic Interactions, Mass Spectrometry, Multivariate Analysis, Temperature.
Abstract
In this study, we describe the optimization of a Hydrophilic Interaction Liquid Chromatography coupled to mass spectrometry (HILIC-MS) method for the evaluation of 14 metabolites related to the de novo synthesis of pyrimidines (dnSP) while using multivariate analysis, which is the metabolic pathway for pyrimidine nucleotide production. A multivariate design was used to set the conditions of the column temperature, flow of the mobile phase, additive concentration, gradient rate, and pH of the mobile phase in order to attain higher peak resolution and ionization efficiency in shorter analysis times. The optimization process was carried out while using factorial fractional designs, Box-Behnken design and central composite design while using two zwitterionic columns, ZIC-p-HILIC and ZIC-HILIC, polymeric, and silica-based columns, respectively. The factors were evaluated while using resolution (R), retention factor (k), efficiency of the column (N), and peak height (h) as the response variables. The best optimized conditions were found with the ZIC-p-HILIC column: elution gradient rate 2 min., pH 7.0, temperature 45 °C, mobile phase flow of 0.35 mL min-1, and additive (ammonium acetate) concentration of 6 mM. The total analysis time was 28 min. The ZIC-p-HILIC LC-MS method yielded satisfactory results for linearity of calibration curves, limit of detection (LOD), and limit of quantification (LOQ). The method has been shown to be appropriate for the analysis of dnSP on samples of tomato plants that were infected with Phytophthora infestans.
DOI: 10.3390/biom9080328
PubMed: 31370321
PubMed Central: PMC6722987
Affiliations:
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Zimmermann</name><affiliation wicri:level="1"><nlm:affiliation>Departamento de Ciencias Biológicas, Universidad de los Andes, Bogotá 111711, Colombia.</nlm:affiliation><country xml:lang="fr">Colombie</country><wicri:regionArea>Departamento de Ciencias Biológicas, Universidad de los Andes, Bogotá 111711</wicri:regionArea><wicri:noRegion>Bogotá 111711</wicri:noRegion></affiliation></author><author><name sortKey="Carazzone, Chiara" sort="Carazzone, Chiara" uniqKey="Carazzone C" first="Chiara" last="Carazzone">Chiara Carazzone</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Advanced Analytical Techniques in Natural Products, Universidad de los Andes, Bogotá 111711, Colombia. c.carazzone@uniandes.edu.co.</nlm:affiliation><country xml:lang="fr">Colombie</country><wicri:regionArea>Laboratory of Advanced Analytical Techniques in Natural Products, Universidad de los Andes, Bogotá 111711</wicri:regionArea><wicri:noRegion>Bogotá 111711</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biomolecules</title><idno type="eISSN">2218-273X</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Chromatography, Liquid (MeSH)</term><term>Hydrogen-Ion Concentration (MeSH)</term><term>Hydrophobic and Hydrophilic Interactions (MeSH)</term><term>Lycopersicon esculentum (metabolism)</term><term>Lycopersicon esculentum (microbiology)</term><term>Mass Spectrometry (MeSH)</term><term>Multivariate Analysis (MeSH)</term><term>Phytophthora infestans (physiology)</term><term>Pyrimidines (metabolism)</term><term>Temperature (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Analyse multifactorielle (MeSH)</term><term>Chromatographie en phase liquide (MeSH)</term><term>Concentration en ions d'hydrogène (MeSH)</term><term>Interactions hydrophobes et hydrophiles (MeSH)</term><term>Lycopersicon esculentum (microbiologie)</term><term>Lycopersicon esculentum (métabolisme)</term><term>Phytophthora infestans (physiologie)</term><term>Pyrimidines (métabolisme)</term><term>Spectrométrie de masse (MeSH)</term><term>Température (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Pyrimidines</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Lycopersicon esculentum</term></keywords><keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Lycopersicon esculentum</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Lycopersicon esculentum</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Lycopersicon esculentum</term><term>Pyrimidines</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Phytophthora infestans</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Phytophthora infestans</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Chromatography, Liquid</term><term>Hydrogen-Ion Concentration</term><term>Hydrophobic and Hydrophilic Interactions</term><term>Mass Spectrometry</term><term>Multivariate Analysis</term><term>Temperature</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse multifactorielle</term><term>Chromatographie en phase liquide</term><term>Concentration en ions d'hydrogène</term><term>Interactions hydrophobes et hydrophiles</term><term>Spectrométrie de masse</term><term>Température</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In this study, we describe the optimization of a Hydrophilic Interaction Liquid Chromatography coupled to mass spectrometry (HILIC-MS) method for the evaluation of 14 metabolites related to the de novo synthesis of pyrimidines (dnSP) while using multivariate analysis, which is the metabolic pathway for pyrimidine nucleotide production. A multivariate design was used to set the conditions of the column temperature, flow of the mobile phase, additive concentration, gradient rate, and pH of the mobile phase in order to attain higher peak resolution and ionization efficiency in shorter analysis times. The optimization process was carried out while using factorial fractional designs, Box-Behnken design and central composite design while using two zwitterionic columns, ZIC-p-HILIC and ZIC-HILIC, polymeric, and silica-based columns, respectively. The factors were evaluated while using resolution (R), retention factor (<i>k</i>), efficiency of the column (N), and peak height (h) as the response variables. The best optimized conditions were found with the ZIC-p-HILIC column: elution gradient rate 2 min., pH 7.0, temperature 45 °C, mobile phase flow of 0.35 mL min<sup>-1</sup>, and additive (ammonium acetate) concentration of 6 mM. The total analysis time was 28 min. The ZIC-p-HILIC LC-MS method yielded satisfactory results for linearity of calibration curves, limit of detection (LOD), and limit of quantification (LOQ). The method has been shown to be appropriate for the analysis of dnSP on samples of tomato plants that were infected with Phytophthora infestans.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">31370321</PMID><DateCompleted><Year>2020</Year><Month>06</Month><Day>02</Day></DateCompleted><DateRevised><Year>2020</Year><Month>06</Month><Day>02</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">2218-273X</ISSN><JournalIssue CitedMedium="Internet"><Volume>9</Volume><Issue>8</Issue><PubDate><Year>2019</Year><Month>07</Month><Day>31</Day></PubDate></JournalIssue><Title>Biomolecules</Title><ISOAbbreviation>Biomolecules</ISOAbbreviation></Journal><ArticleTitle>Hydrophilic Interaction Liquid Chromatography Coupled to Mass Spectrometry and Multivariate Analysis of the <i>De Novo</i> Pyrimidine Pathway Metabolites.</ArticleTitle><ELocationID EIdType="pii" ValidYN="Y">E328</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.3390/biom9080328</ELocationID><Abstract><AbstractText>In this study, we describe the optimization of a Hydrophilic Interaction Liquid Chromatography coupled to mass spectrometry (HILIC-MS) method for the evaluation of 14 metabolites related to the de novo synthesis of pyrimidines (dnSP) while using multivariate analysis, which is the metabolic pathway for pyrimidine nucleotide production. A multivariate design was used to set the conditions of the column temperature, flow of the mobile phase, additive concentration, gradient rate, and pH of the mobile phase in order to attain higher peak resolution and ionization efficiency in shorter analysis times. The optimization process was carried out while using factorial fractional designs, Box-Behnken design and central composite design while using two zwitterionic columns, ZIC-p-HILIC and ZIC-HILIC, polymeric, and silica-based columns, respectively. The factors were evaluated while using resolution (R), retention factor (<i>k</i>), efficiency of the column (N), and peak height (h) as the response variables. The best optimized conditions were found with the ZIC-p-HILIC column: elution gradient rate 2 min., pH 7.0, temperature 45 °C, mobile phase flow of 0.35 mL min<sup>-1</sup>, and additive (ammonium acetate) concentration of 6 mM. The total analysis time was 28 min. The ZIC-p-HILIC LC-MS method yielded satisfactory results for linearity of calibration curves, limit of detection (LOD), and limit of quantification (LOQ). The method has been shown to be appropriate for the analysis of dnSP on samples of tomato plants that were infected with Phytophthora infestans.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Galeano Garcia</LastName><ForeName>Paula</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Laboratory of Advanced Analytical Techniques in Natural Products, Universidad de los Andes, Bogotá 111711, Colombia. plgaleanog@gmail.com.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>"Grupo de Investigación en Productos Naturales Amazónicos", Facultad de Ciencias Básicas, Universidad de la Amazonia, Florencia 180002, Colombia. plgaleanog@gmail.com.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zimmermann</LastName><ForeName>Barbara H</ForeName><Initials>BH</Initials><AffiliationInfo><Affiliation>Departamento de Ciencias Biológicas, Universidad de los Andes, Bogotá 111711, Colombia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Carazzone</LastName><ForeName>Chiara</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Laboratory of Advanced Analytical Techniques in Natural Products, Universidad de los Andes, Bogotá 111711, Colombia. c.carazzone@uniandes.edu.co.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>07</Month><Day>31</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Biomolecules</MedlineTA><NlmUniqueID>101596414</NlmUniqueID><ISSNLinking>2218-273X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011743">Pyrimidines</NameOfSubstance></Chemical><Chemical><RegistryNumber>K8CXK5Q32L</RegistryNumber><NameOfSubstance UI="C030986">pyrimidine</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D002853" MajorTopicYN="Y">Chromatography, Liquid</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057927" MajorTopicYN="Y">Hydrophobic and Hydrophilic Interactions</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018551" MajorTopicYN="N">Lycopersicon esculentum</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName><QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013058" MajorTopicYN="Y">Mass Spectrometry</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015999" MajorTopicYN="N">Multivariate Analysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D055750" MajorTopicYN="N">Phytophthora infestans</DescriptorName><QualifierName UI="Q000502" MajorTopicYN="N">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011743" MajorTopicYN="N">Pyrimidines</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013696" MajorTopicYN="N">Temperature</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Box–Behnken design</Keyword><Keyword MajorTopicYN="Y">Phytophthora infestans</Keyword><Keyword MajorTopicYN="Y">central composite design</Keyword><Keyword MajorTopicYN="Y">experimental design</Keyword><Keyword MajorTopicYN="Y">hydrophilic interaction liquid chromatography</Keyword><Keyword MajorTopicYN="Y">plant-pathogen interaction</Keyword><Keyword MajorTopicYN="Y">tomato plants</Keyword><Keyword MajorTopicYN="Y">zwitterionic column</Keyword></KeywordList><CoiStatement>The authors declare no conflict of interest.</CoiStatement></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>06</Month><Day>23</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2019</Year><Month>07</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>07</Month><Day>18</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>8</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>8</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>6</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId 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IdType="pubmed">17931636</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Colombie</li></country></list><tree><country name="Colombie"><noRegion><name sortKey="Galeano Garcia, Paula" sort="Galeano Garcia, Paula" uniqKey="Galeano Garcia P" first="Paula" last="Galeano Garcia">Paula Galeano Garcia</name></noRegion><name sortKey="Carazzone, Chiara" sort="Carazzone, Chiara" uniqKey="Carazzone C" first="Chiara" last="Carazzone">Chiara Carazzone</name><name sortKey="Galeano Garcia, Paula" sort="Galeano Garcia, Paula" uniqKey="Galeano Garcia P" first="Paula" last="Galeano Garcia">Paula Galeano Garcia</name><name sortKey="Zimmermann, Barbara H" sort="Zimmermann, Barbara H" uniqKey="Zimmermann B" first="Barbara H" last="Zimmermann">Barbara H. 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