Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization.
Identifieur interne : 001E62 ( Main/Exploration ); précédent : 001E61; suivant : 001E63Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization.
Auteurs : Heather L. Lord [Canada] ; Jack Rosenfeld ; Vitaly Volovich ; Dinesh Kumbhare ; Bill ParkinsonSource :
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [ 1873-376X ] ; 2009.
English descriptors
- KwdEn :
- MESH :
- chemical , blood : Biomarkers, Malondialdehyde.
- methods : Chromatography, Liquid, Mass Spectrometry.
- Automation, Humans, Oxidative Stress, Spectrometry, Fluorescence, Spectrophotometry, Ultraviolet.
Abstract
Analytical derivatization (AD) increases the sensitivity of analysis by one to three orders of magnitude, stabilizes labile analytes and converts them into readily extractable products. Using a variant of this technique, we applied solid phase analytical derivatization (SPAD) to fully automate extraction, derivatization and liquid chromatography. The resulting device (AutoSPAD) determined malonyldialdehyde (MDA) from biological fluids. This biomarker of oxidative stress is highly water-soluble (500 g/L at pH 7), chemically labile and lacks any functionality that enables detection at high sensitivity. AutoSPAD utilizes column-switching technology to load DANSYL hydrazine onto the solid phase, pass the biological sample over the resulting reactor bed for derivatization on the surface to form a hydrophobic derivative suitable for increasing sensitivity of any other LC technique including LC-MS/MS. The hydrophobic solid phase retains the derivative during washing steps, following which AutoSPAD transfers the derivatized extract to the analytical column for separation and detection by fluorescence. In plasma, however, MDA exists both in free form and covalently bound to protein. Measuring MDA from plasma, therefore, required identification of appropriate protein precipitation and hydrolysis conditions. Under these conditions, the DANSYL derivative formed at only one aldehydic position but did not cyclize as reported for other reactions between hydrazine reagents and MDA. The calibration curve using approximately 7 microL of plasma was linear (r(2)=0.999) in the physiological range (0.1-3 microg/mL) and the relative standard deviation of replicate determinations at 1 microg/mL was less than 5%.
DOI: 10.1016/j.jchromb.2008.12.035
PubMed: 19124277
Affiliations:
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Lord</name><affiliation wicri:level="4"><nlm:affiliation>Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON</wicri:regionArea><orgName type="university">Université McMaster</orgName><placeName><settlement type="city">Hamilton (Ontario)</settlement><region type="state">Ontario</region></placeName></affiliation></author><author><name sortKey="Rosenfeld, Jack" sort="Rosenfeld, Jack" uniqKey="Rosenfeld J" first="Jack" last="Rosenfeld">Jack Rosenfeld</name></author><author><name sortKey="Volovich, Vitaly" sort="Volovich, Vitaly" uniqKey="Volovich V" first="Vitaly" last="Volovich">Vitaly Volovich</name></author><author><name sortKey="Kumbhare, Dinesh" sort="Kumbhare, Dinesh" uniqKey="Kumbhare D" first="Dinesh" last="Kumbhare">Dinesh Kumbhare</name></author><author><name sortKey="Parkinson, Bill" sort="Parkinson, Bill" uniqKey="Parkinson B" first="Bill" last="Parkinson">Bill Parkinson</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19124277</idno><idno type="pmid">19124277</idno><idno type="doi">10.1016/j.jchromb.2008.12.035</idno><idno type="wicri:Area/PubMed/Corpus">000E77</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000E77</idno><idno type="wicri:Area/PubMed/Curation">000E77</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000E77</idno><idno type="wicri:Area/PubMed/Checkpoint">000E77</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000E77</idno><idno type="wicri:Area/Ncbi/Merge">000A08</idno><idno type="wicri:Area/Ncbi/Curation">000A08</idno><idno type="wicri:Area/Ncbi/Checkpoint">000A08</idno><idno type="wicri:Area/Main/Merge">002015</idno><idno type="wicri:Area/Main/Curation">001E62</idno><idno type="wicri:Area/Main/Exploration">001E62</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization.</title><author><name sortKey="Lord, Heather L" sort="Lord, Heather L" uniqKey="Lord H" first="Heather L" last="Lord">Heather L. Lord</name><affiliation wicri:level="4"><nlm:affiliation>Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON</wicri:regionArea><orgName type="university">Université McMaster</orgName><placeName><settlement type="city">Hamilton (Ontario)</settlement><region type="state">Ontario</region></placeName></affiliation></author><author><name sortKey="Rosenfeld, Jack" sort="Rosenfeld, Jack" uniqKey="Rosenfeld J" first="Jack" last="Rosenfeld">Jack Rosenfeld</name></author><author><name sortKey="Volovich, Vitaly" sort="Volovich, Vitaly" uniqKey="Volovich V" first="Vitaly" last="Volovich">Vitaly Volovich</name></author><author><name sortKey="Kumbhare, Dinesh" sort="Kumbhare, Dinesh" uniqKey="Kumbhare D" first="Dinesh" last="Kumbhare">Dinesh Kumbhare</name></author><author><name sortKey="Parkinson, Bill" sort="Parkinson, Bill" uniqKey="Parkinson B" first="Bill" last="Parkinson">Bill Parkinson</name></author></analytic><series><title level="j">Journal of chromatography. B, Analytical technologies in the biomedical and life sciences</title><idno type="eISSN">1873-376X</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Automation</term><term>Biomarkers (blood)</term><term>Chromatography, Liquid (methods)</term><term>Humans</term><term>Malondialdehyde (blood)</term><term>Mass Spectrometry (methods)</term><term>Oxidative Stress</term><term>Spectrometry, Fluorescence</term><term>Spectrophotometry, Ultraviolet</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Biomarkers</term><term>Malondialdehyde</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Chromatography, Liquid</term><term>Mass Spectrometry</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Automation</term><term>Humans</term><term>Oxidative Stress</term><term>Spectrometry, Fluorescence</term><term>Spectrophotometry, Ultraviolet</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Analytical derivatization (AD) increases the sensitivity of analysis by one to three orders of magnitude, stabilizes labile analytes and converts them into readily extractable products. Using a variant of this technique, we applied solid phase analytical derivatization (SPAD) to fully automate extraction, derivatization and liquid chromatography. The resulting device (AutoSPAD) determined malonyldialdehyde (MDA) from biological fluids. This biomarker of oxidative stress is highly water-soluble (500 g/L at pH 7), chemically labile and lacks any functionality that enables detection at high sensitivity. AutoSPAD utilizes column-switching technology to load DANSYL hydrazine onto the solid phase, pass the biological sample over the resulting reactor bed for derivatization on the surface to form a hydrophobic derivative suitable for increasing sensitivity of any other LC technique including LC-MS/MS. The hydrophobic solid phase retains the derivative during washing steps, following which AutoSPAD transfers the derivatized extract to the analytical column for separation and detection by fluorescence. In plasma, however, MDA exists both in free form and covalently bound to protein. Measuring MDA from plasma, therefore, required identification of appropriate protein precipitation and hydrolysis conditions. Under these conditions, the DANSYL derivative formed at only one aldehydic position but did not cyclize as reported for other reactions between hydrazine reagents and MDA. The calibration curve using approximately 7 microL of plasma was linear (r(2)=0.999) in the physiological range (0.1-3 microg/mL) and the relative standard deviation of replicate determinations at 1 microg/mL was less than 5%.</div></front></TEI><affiliations><list><country><li>Canada</li></country><region><li>Ontario</li></region><settlement><li>Hamilton (Ontario)</li></settlement><orgName><li>Université McMaster</li></orgName></list><tree><noCountry><name sortKey="Kumbhare, Dinesh" sort="Kumbhare, Dinesh" uniqKey="Kumbhare D" first="Dinesh" last="Kumbhare">Dinesh Kumbhare</name><name sortKey="Parkinson, Bill" sort="Parkinson, Bill" uniqKey="Parkinson B" first="Bill" last="Parkinson">Bill Parkinson</name><name sortKey="Rosenfeld, Jack" sort="Rosenfeld, Jack" uniqKey="Rosenfeld J" first="Jack" last="Rosenfeld">Jack Rosenfeld</name><name sortKey="Volovich, Vitaly" sort="Volovich, Vitaly" uniqKey="Volovich V" first="Vitaly" last="Volovich">Vitaly Volovich</name></noCountry><country name="Canada"><region name="Ontario"><name sortKey="Lord, Heather L" sort="Lord, Heather L" uniqKey="Lord H" first="Heather L" last="Lord">Heather L. 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