title: Development and evaluation of a liquid chromatography-mass spectrometry method for rapid, accurate quantitation of malondialdehyde in human plasma.
abstract: Malondialdhyde (MDA) is a commonly used marker of lipid peroxidation in oxidative stress. To provide a sensitive analytical method that is compatible with high throughput, we developed a multiple reaction monitoring-mass spectrometry (MRM-MS) approach using 3-nitrophenylhydrazine chemical derivatization, isotope-labeling, and liquid chromatography (LC) with electrospray ionization (ESI)-tandem mass spectrometry assay to accurately quantify MDA in human plasma. A stable isotope-labeled internal standard was used to compensate for ESI matrix effects. The assay is linear (R(2)=0.9999) over a 20,000-fold concentration range with a lower limit of quantitation of 30fmol (on-column). Intra- and inter-run coefficients of variation (CVs) were <2% and ∼10% respectively. The derivative was stable for >36h at 5°C. Standards spiked into plasma had recoveries of 92-98%. When compared to a common LC-UV method, the LC-MS method found near-identical MDA concentrations. A pilot project to quantify MDA in patient plasma samples (n=26) in a study of major depressive disorder with winter-type seasonal pattern (MDD-s) confirmed known associations between MDA concentrations and obesity (p<0.02). The LC-MS method provides high sensitivity and high reproducibility for quantifying MDA in human plasma. The simple sample preparation and rapid analysis time (5x faster than LC-UV) offers high throughput for large-scale clinical applications.
authors.affiliation: University of Victoria-Genome BC Proteomics Centre, Vancouver Island Technology Park, 3101-4464 Markham Street, Victoria, BC V8Z 7X8, Canada; Department of Biochemistry and Microbiology, University of Victoria, Petch Building Room 207, 3800 Finnerty Road, Victoria, BC V8P 5C2, Canada.
authors.forename: Constance A
authors.initials: CA
authors.lastname: Sobsey
authors.affiliation: University of Victoria-Genome BC Proteomics Centre, Vancouver Island Technology Park, 3101-4464 Markham Street, Victoria, BC V8Z 7X8, Canada.
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.affiliation: University of Victoria-Genome BC Proteomics Centre, Vancouver Island Technology Park, 3101-4464 Markham Street, Victoria, BC V8Z 7X8, Canada.
authors.forename: Karen
authors.initials: K
authors.lastname: Lin
authors.affiliation: Pharmacology and Toxicology Department, Sunnybrook Health Sciences Centre, 2075 Bayview Ave., Toronto, ON M4N 3M5, Canada.
authors.forename: Walter
authors.initials: W
authors.lastname: Swardfager
authors.affiliation: Department of Psychiatry, Sunnybrook Health Sciences Centre, 2075 Bayview Ave., Toronto, ON M4N 3M5, Canada.
authors.forename: Anthony
authors.initials: A
authors.lastname: Levitt
authors.affiliation: University of Victoria-Genome BC Proteomics Centre, Vancouver Island Technology Park, 3101-4464 Markham Street, Victoria, BC V8Z 7X8, Canada; Department of Biochemistry and Microbiology, University of Victoria, Petch Building Room 207, 3800 Finnerty Road, Victoria, BC V8P 5C2, Canada; Segal Cancer Centre, Jewish General Hospital, McGill University, 3755 Cote Ste-Catherine Road, Montreal, QC H3T 1E2, Canada. Electronic address: christoph@proteincentre.com.
authors.forename: Christoph H
authors.initials: CH
authors.lastname: Borchers
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003865:Major Depressive Disorder / Q000097:blood; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008315:Malondialdehyde / Q000097:blood*; D010659:Phenylhydrazines / Q000737:chemistry; D010865:Pilot Projects; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: C523491:3-nitrophenylhydrazine; D010659:Phenylhydrazines; D008315:Malondialdehyde
keywords: 3-Nitrophenylhydrazine; Derivatization; Isotope-labeling; LC/MRM–MS; Malondialdehyde; Plasma