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Published on: March 23, 2022
Quantitation of cardiolipin molecular species in spontaneously hypertensive heart failure rats using electrospray
Genevieve C Sparagna1, Chris A Johnson, Sylvia A McCune
1Department of Integrative Physiology, University of Colorado Cardiovascular Institute, University of Colorado at Boulder, Boulder, CO 80309-0354, USA. genevieve.sparagna@colorado.edu
Insights
Quantifying cardiolipin (CL) molecular species in heart failure is crucial. This study developed a method using mass spectrometry and an internal standard to accurately measure CL changes in failing rat hearts.
Area of Science:
- Biochemistry
- Cardiovascular Science
- Analytical Chemistry
Background:
- Cardiolipin (CL) is vital for mitochondrial function and cardiac health.
- Previous studies primarily used mass spectrometry for qualitative CL analysis.
- Quantitative analysis of CL molecular species in heart failure is lacking.
Purpose of the Study:
- To develop and validate a quantitative method for analyzing cardiac cardiolipin (CL) molecular species.
- To investigate changes in individual CL molecular species in a rat model of heart failure.
- To assess the feasibility of CL quantification in both mitochondrial and tissue samples.
Main Methods:
- Utilized electrospray ionization mass spectrometry (ESI-MS) for CL analysis.
- Employed 1,1',2,2'-tetramyristoyl cardiolipin as an internal standard for quantification.
- Applied tandem mass spectrometry (MS/MS) to identify acyl chain composition of CL species.
Main Results:
- Established a linear relationship for quantitative CL measurement using the internal standard.
- Demonstrated successful extraction and quantification of CL from cardiac mitochondria and left ventricular tissue.
- Observed a significant decrease in the major tetralinoloyl CL species and a significant increase in minor CL species in heart failure models.
Conclusions:
- The developed ESI-MS method with an internal standard enables accurate quantification of cardiac CL molecular species.
- This quantitative approach reveals specific alterations in CL composition during heart failure.
- Findings provide insights into the role of CL remodeling in cardiac dysfunction.
Abstract:
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.
