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Analysis of sterols by mass spectrometry is a fundamental technique allowing for both qualitative and quantitative characterization of sterol molecular lipid species. Lipids are isolated from matrix or matrices by homogenization and solvent extraction, and converted into species amenable for ionization either by derivatization or adduct formation. Chromatogaphy (either gas or liquid phase) can assist with the resolution of sterols. Tandem mass spectrometry allows the precise identification of sterol lipid species, while comparison to internal standards added during extraction enables accurate quantification.

Excitotoxicity due to excess stimulation of glutamate receptors in neurons is accompanied by increased Ca(2+) influx, stimulation of Ca(2+)-dependent enzymes, ATP depletion, increase in lipid peroxidation products, and loss of glutathione. These changes resemble neurochemical alterations in acute neuronal injury (stroke, spinal cord injury, and traumatic brain injury) and chronic neurodegenerative diseases such as Alzheimer's disease. Intracerebroventricular injection of the potent glutamate analog kainate in rats results in increased cholesterol concentration in the hippocampus at short to medium time intervals, i.e., 3 days-1 week post-injection, as detected by gas chromatography-mass spectrometry in the lesioned hippocampus. This is accompanied by an early increase in levels of cholesterol biosynthetic precursors and increases in both enzymatically derived oxysterols such as 24-hydroxycholesterol and cholesterol oxidation products (COPs) generated by reactive oxygen species, including cholesterol epoxides and 7-ketocholesterol. In contrast to COPs, no change in concentration of the neurosteroid pregnenolone was found after KA injury. Cholesterol and COPs significantly increase exocytosis in cultured PC12 cells and neurons, and both oxysterols and COPs are able to induce cytotoxic and apoptotic injuries in different cell types, including neurons. Together, the findings suggest that increased cholesterol and COPs after KA excitotoxicity could themselves lead to disturbed neuronal ion homeostasis, increased neurotransmitter release, and propagation of excitotoxicity.