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Archana A. Gupta, Katyayini Aribindi, Anna K. Junk, Sanjoy K. Bhattacharya; Mass Spectrometric Lipidomic Analyses Of Psychosine And Glycolipids Of Trabecular Meshwork. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3831.
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© ARVO (1962-2015); The Authors (2016-present)
To identify the endogenous Psychosine and glycolipids of the trabecular meshwork (TM) by mass spectrometric shotgun lipidomics.
The human TM samples were collected adhering to tenets of declaration of Helsinki under IRB approved protocols. The TM were obtained from Caucasian donors with mean age of 60 ± 6 years (n=8 each). Control TM samples were from cadavers and left over rim tissues of penetrating keratoplasty surgery respectively. Lipids were extracted using the Bligh and Dyer method, and re-suspended in Isopropyl, Acetonitrile solution. For lipid analyses we have used positive ion mode neutral loss scan (NLS) for psychosine (neutral product loss of m/z 180) using a collision energy of 24 and spray voltage of 4000V with a TSQ Quantum Access Max instrument. The samples were infused and scanned for one minute between 200 m/z to 1000 m/z. Ratiometric quantification was performed using quantitative galactosyl sphingosine D-galactosyl-ß1-1'-D-erythro-sphingosine (parent mass 461.64).
We found several lipid species in the human TM that are identified by NLS scan for neutral loss of m/z 180. We found a number of glycolipids (sugar moiety: neutral loss of m/z 180) that were identified in scan of m/z 180 neutral loss. Unique glycolipids in controls that are absent in glaucomatous TM are being subjected to further characterization using high-resolution mass spectrometry.
We found unique glycolipids in control TM that are either absent or present in very low (sub picomole) levels in glaucomatous samples.
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