Illawara Health and Medical Research Institute and School of Health Sciences, University of Wollongong, Wollongong, New South Wales, Australia.
Invest Ophthalmol Vis Sci. 2013 Nov 11;54(12):7417-24. doi: 10.1167/iovs.13-12916.
To quantify the molecular lipid composition of patient-matched tear and meibum samples and compare tear and meibum lipid molecular profiles.
Lipids were extracted from tears and meibum by bi-phasic methods using 10:3 tert-butyl methyl ether:methanol, washed with aqueous ammonium acetate, and analyzed by chip-based nanoelectrospray ionization tandem mass spectrometry. Targeted precursor ion and neutral loss scans identified individual molecular lipids and quantification was obtained by comparison to internal standards in each lipid class.
Two hundred and thirty-six lipid species were identified and quantified from nine lipid classes comprised of cholesterol esters, wax esters, (O-acyl)-ω-hydroxy fatty acids, triacylglycerols, phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, sphingomyelin, and phosphatidylserine. With the exception of phospholipids, lipid molecular profiles were strikingly similar between tears and meibum.
Comparisons between tears and meibum indicate that meibum is likely to supply the majority of lipids in the tear film lipid layer. However, the observed higher mole ratio of phospholipid in tears shows that analysis of meibum alone does not provide a complete understanding of the tear film lipid composition.
定量分析患者匹配的泪液和睑脂样本中的分子脂质组成,并比较泪液和睑脂的脂质分子谱。
采用 10:3 叔丁基甲基醚-甲醇双相法从泪液和睑脂中提取脂质,用乙酸铵水溶液洗涤,并用基于芯片的纳喷雾电离串联质谱分析。靶向前体离子和中性丢失扫描鉴定了各个分子脂质,并通过与每个脂质类别的内标进行比较来定量。
从胆固醇酯、蜡酯、(O-酰基)-ω-羟基脂肪酸、三酰甘油、磷脂酰胆碱、溶血磷脂酰胆碱、磷脂酰乙醇胺、鞘磷脂和磷脂酰丝氨酸等 9 个脂质类中鉴定并定量了 236 种脂质。除了磷脂外,泪液和睑脂的脂质分子谱非常相似。
泪液和睑脂的比较表明,睑脂可能为泪膜脂质层提供了大部分脂质。然而,观察到泪液中磷脂的摩尔比更高,表明仅分析睑脂并不能全面了解泪膜脂质的组成。