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Journal of Lipid Research, Vol. 46, 1196-1204, June 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Quantitation of cardiolipin molecular species in spontaneously hypertensive heart failure rats using electrospray ionization mass spectrometry

Genevieve C. Sparagna^1,*, Chris A. Johnson, Sylvia A. McCune^*, Russell L. Moore^* and Robert C. Murphy

^* Department of Integrative Physiology, University of Colorado Cardiovascular Institute, University of Colorado at Boulder, Boulder, CO 80309-0354
Department of Pharmacology, University of Colorado Health Sciences Center, Aurora, CO 80045-0511

Published, JLR Papers in Press, March 16, 2005. DOI 10.1194/jlr.M500031-JLR200

¹ To whom correspondence should be addressed. e-mail: genevieve.sparagna{at}colorado.edu

Electrospray ionization mass spectrometry has previously been used to probe qualitative changes in the phospholipid cardiolipin (CL), but it has rarely been used in a quantitative manner. We assessed changes in the amount of individual molecular species of cardiac CL present in a model of congestive heart failure using 1,1',2,2'-tetramyristoyl cardiolipin as an internal standard. There was a linear relationship between the ratio of the negative molecular ion ([M-H]^–) current from four different CL reference standards and the [M-H]^– from the internal standard, as a function of the concentration of CL molecular species. Therefore, this internal standard can be used to quantitate many naturally occurring CL molecular species over a wide range of CL concentrations. Using this method, changes to individual molecular species of CL in failing hearts from male spontaneously hypertensive heart failure rats were examined. CL isolated from cardiac mitochondria was compared with left ventricular tissue to demonstrate the feasibility of extracting and quantitating CL from either mitochondrial or tissue samples.

The acyl chain composition of individual CL molecular species was identified using tandem mass spectrometry. In animals with heart failure, the major cardiac CL species (tetralinoloyl) decreased significantly, whereas other minor CL species were significantly increased.

Abbreviations: CL, cardiolipin; ESI, electrospray ionization; LC-MS, liquid chromatography-mass spectrometry; [M-2H]^2–, doubly charged negative molecular ion; [M-H]^–, negative molecular ion; M₄CL, 1,1', 2,2'-tetramyristoyl cardiolipin; MS/MS, tandem collisional mass spectrometry; O₄CL, cardiolipin with four oleates; SHHF, spontaneously hypertensive heart failure

Supplementary key words mitochondria • cardiac • internal standard • tandem mass spectrometry

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