Analysis of acrylamido-buffers for isoelectric focusing by capillary zone electrophoresis.
Identifieur interne : 000757 ( Ncbi/Merge ); précédent : 000756; suivant : 000758Analysis of acrylamido-buffers for isoelectric focusing by capillary zone electrophoresis.
Auteurs : P G Righetti [Italie] ; C. Ettori ; M. ChiariSource :
- Electrophoresis [ 0173-0835 ] ; 1991.
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Abstract
Immobilized pH gradients use a series of weak acrylamido acids and bases (Immobiline) to create a pH gradient along the separation axis. These buffers can be degraded in water by two mechanisms: (i) hydrolysis of the amido bond, with generation of free acrylic acid and either an amino acid or a diamine; (ii) autopolymerization to oligomers and/or n-mers. In order to check for these degradation products, different capillary zone electrophoresis systems for analysis of all Immobilines have been devised. The acidic compounds are resolved in 100 mM acetate, pH 4.0, whereas the alkaline Immobilines are separated in 50 mM phosphate buffer, pH 7.7 (or pH 7.2 for the weaker species). Polymers of alkaline Immobilines are resolved in 50 mM phosphate buffer, pH 2.5, in 1% Ficoll-400. All Immobilines are detected underivatized, by their adsorption at 214 or 254 nm. A calibration curve has been constructed for quantification of acrylic acid contamination. As little as 1 mol% of acrylic acid contamination in Immobiline solutions can be detected, with a sensitivity limit below 0.2 mM (at the injection port).
DOI: 10.1002/elps.1150120110
PubMed: 2050100
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Ettori</name></author><author><name sortKey="Chiari, M" sort="Chiari, M" uniqKey="Chiari M" first="M" last="Chiari">M. Chiari</name></author></analytic><series><title level="j">Electrophoresis</title><idno type="ISSN">0173-0835</idno><imprint><date when="1991" type="published">1991</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Acrylamides</term><term>Calibration</term><term>Electrophoresis</term><term>Hydrogen-Ion Concentration</term><term>Hydrolysis</term><term>Isoelectric Focusing</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acrylamides</term><term>Calibrage</term><term>Concentration en ions d'hydrogène</term><term>Focalisation isoélectrique</term><term>Hydrolyse</term><term>Sensibilité et spécificité</term><term>Électrophorèse</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Acrylamides</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Calibration</term><term>Electrophoresis</term><term>Hydrogen-Ion Concentration</term><term>Hydrolysis</term><term>Isoelectric Focusing</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Acrylamides</term><term>Calibrage</term><term>Concentration en ions d'hydrogène</term><term>Focalisation isoélectrique</term><term>Hydrolyse</term><term>Sensibilité et spécificité</term><term>Électrophorèse</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Immobilized pH gradients use a series of weak acrylamido acids and bases (Immobiline) to create a pH gradient along the separation axis. These buffers can be degraded in water by two mechanisms: (i) hydrolysis of the amido bond, with generation of free acrylic acid and either an amino acid or a diamine; (ii) autopolymerization to oligomers and/or n-mers. In order to check for these degradation products, different capillary zone electrophoresis systems for analysis of all Immobilines have been devised. The acidic compounds are resolved in 100 mM acetate, pH 4.0, whereas the alkaline Immobilines are separated in 50 mM phosphate buffer, pH 7.7 (or pH 7.2 for the weaker species). Polymers of alkaline Immobilines are resolved in 50 mM phosphate buffer, pH 2.5, in 1% Ficoll-400. All Immobilines are detected underivatized, by their adsorption at 214 or 254 nm. A calibration curve has been constructed for quantification of acrylic acid contamination. As little as 1 mol% of acrylic acid contamination in Immobiline solutions can be detected, with a sensitivity limit below 0.2 mM (at the injection port).</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">2050100</PMID><DateCompleted><Year>1991</Year><Month>07</Month><Day>23</Day></DateCompleted><DateRevised><Year>2006</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0173-0835</ISSN><JournalIssue CitedMedium="Print"><Volume>12</Volume><Issue>1</Issue><PubDate><Year>1991</Year><Month>Jan</Month></PubDate></JournalIssue><Title>Electrophoresis</Title><ISOAbbreviation>Electrophoresis</ISOAbbreviation></Journal><ArticleTitle>Analysis of acrylamido-buffers for isoelectric focusing by capillary zone electrophoresis.</ArticleTitle><Pagination><MedlinePgn>55-8</MedlinePgn></Pagination><Abstract><AbstractText>Immobilized pH gradients use a series of weak acrylamido acids and bases (Immobiline) to create a pH gradient along the separation axis. These buffers can be degraded in water by two mechanisms: (i) hydrolysis of the amido bond, with generation of free acrylic acid and either an amino acid or a diamine; (ii) autopolymerization to oligomers and/or n-mers. In order to check for these degradation products, different capillary zone electrophoresis systems for analysis of all Immobilines have been devised. The acidic compounds are resolved in 100 mM acetate, pH 4.0, whereas the alkaline Immobilines are separated in 50 mM phosphate buffer, pH 7.7 (or pH 7.2 for the weaker species). Polymers of alkaline Immobilines are resolved in 50 mM phosphate buffer, pH 2.5, in 1% Ficoll-400. All Immobilines are detected underivatized, by their adsorption at 214 or 254 nm. A calibration curve has been constructed for quantification of acrylic acid contamination. As little as 1 mol% of acrylic acid contamination in Immobiline solutions can be detected, with a sensitivity limit below 0.2 mM (at the injection port).</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Righetti</LastName><ForeName>P G</ForeName><Initials>PG</Initials><AffiliationInfo><Affiliation>Department of Biomedical Sciences and Technologies, University of Milano, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ettori</LastName><ForeName>C</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Chiari</LastName><ForeName>M</ForeName><Initials>M</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Electrophoresis</MedlineTA><NlmUniqueID>8204476</NlmUniqueID><ISSNLinking>0173-0835</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000178">Acrylamides</NameOfSubstance></Chemical><Chemical><RegistryNumber>88385-55-1</RegistryNumber><NameOfSubstance UI="C059413">Immobiline</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000178" MajorTopicYN="Y">Acrylamides</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002138" MajorTopicYN="N">Calibration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004586" MajorTopicYN="Y">Electrophoresis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006868" MajorTopicYN="N">Hydrolysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007525" MajorTopicYN="Y">Isoelectric Focusing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1991</Year><Month>1</Month><Day>1</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1991</Year><Month>1</Month><Day>1</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1991</Year><Month>1</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">2050100</ArticleId><ArticleId IdType="doi">10.1002/elps.1150120110</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Italie</li></country></list><tree><noCountry><name sortKey="Chiari, M" sort="Chiari, M" uniqKey="Chiari M" first="M" last="Chiari">M. Chiari</name><name sortKey="Ettori, C" sort="Ettori, C" uniqKey="Ettori C" first="C" last="Ettori">C. Ettori</name></noCountry><country name="Italie"><noRegion><name sortKey="Righetti, P G" sort="Righetti, P G" uniqKey="Righetti P" first="P G" last="Righetti">P G Righetti</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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