{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,1,21]],"date-time":"2024-01-21T12:58:33Z","timestamp":1705841913267},"reference-count":38,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2012,2,6]],"date-time":"2012-02-06T00:00:00Z","timestamp":1328486400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Alcoholism Clin &amp;amp; Exp Res"],"published-print":{"date-parts":[[2012,6]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\"><jats:italic>N<\/jats:italic><\/jats:styled-content><jats:sup>\u03b5<\/jats:sup>\u2010ethyllysine (<jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content>) is a major stable adduct formed by the reaction of acetaldehyde (AA) with lysine residues in proteins. However, its occurrence and levels in biological specimens and its relationship with AA\/alcohol exposure\u2010associated disorders have not been fully elucidated. In this study, we have developed a sensitive and specific method to quantitate <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> levels in human plasma proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The method consists of (1) purification of the protein fraction of interest by <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>ephadex <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>\u201015 to remove low molecular substances, (2) hydrolysis of proteins with <jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>ronase <jats:styled-content style=\"fixed-case\">E<\/jats:styled-content> in the presence of stable isotope\u2013labeled internal standards, (3) derivatization of amino acids with pentafluorobenzyl (<jats:styled-content style=\"fixed-case\">PFB<\/jats:styled-content>) bromide, and (4) quantification of the <jats:styled-content style=\"fixed-case\">PFB<\/jats:styled-content> derivatives of <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> and <jats:sc>l<\/jats:sc>\u2010lysine using gas chromatography\u2013negative ion chemical ionization\/mass spectrometry in a selected ion monitoring mode.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Using the above method, the <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> levels in human plasma proteins obtained from 10 each of control subjects and alcoholic patients were measured. <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> was detected in all samples analyzed, the average level of <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> in the plasma proteins of alcoholic patients (1.17\u00a0\u00b1\u00a00.36 <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content>\/1,000 <jats:sc>l<\/jats:sc>\u2010lysine) being significantly higher than that of control subjects (0.26\u00a0\u00b1\u00a00.07 <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content>\/1,000 <jats:sc>l<\/jats:sc>\u2010lysine).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The method could be applied to molecular epidemiological studies to investigate possible associations between the <jats:styled-content style=\"fixed-case\">NEL<\/jats:styled-content> levels in human tissue proteins and human diseases associated with exposure to AA and alcohol.<\/jats:p><\/jats:sec>","DOI":"10.1111\/j.1530-0277.2011.01705.x","type":"journal-article","created":{"date-parts":[[2012,2,7]],"date-time":"2012-02-07T03:12:09Z","timestamp":1328584329000},"page":"1013-1020","source":"Crossref","is-referenced-by-count":5,"title":["Analysis of <scp><i>N<\/i><\/scp><sup>\u03b5<\/sup>\u2010Ethyllysine in Human Plasma Proteins by Gas Chromatography\u2013Negative Ion Chemical Ionization\/Mass Spectrometry as a Biomarker for Exposure to Acetaldehyde and 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