Document Detail

Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization.
MedLine Citation:
PMID:  19124277     Owner:  NLM     Status:  MEDLINE    
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%.
Heather L Lord; Jack Rosenfeld; Vitaly Volovich; Dinesh Kumbhare; Bill Parkinson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-12-24
Journal Detail:
Title:  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences     Volume:  877     ISSN:  1873-376X     ISO Abbreviation:  J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-04-13     Completed Date:  2009-05-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101139554     Medline TA:  J Chromatogr B Analyt Technol Biomed Life Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1292-8     Citation Subset:  IM    
Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.
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MeSH Terms
Biological Markers / blood*
Chromatography, Liquid / methods*
Malondialdehyde / blood*
Mass Spectrometry / methods*
Oxidative Stress
Spectrometry, Fluorescence
Spectrophotometry, Ultraviolet
Reg. No./Substance:
0/Biological Markers; 542-78-9/Malondialdehyde

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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