A method for determination from high density lipoproteins with TOF-MS of phosphatidylethanol by reversed-phase HPLC detection
A method for determination from high density lipoproteins with TOF-MS of phosphatidylethanol by reversed-phase HPLC detection
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DOI：
10.1016/j.ab.2005.03.001
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发表时间：
2005-06-01
影响因子：
2.9
通讯作者：
Lehto, TM
中科院分区：
文献类型：
--
作者：
Tolonen, A;Lehto, TM;Lehto, TM
磷脂酰乙醇 (PEth) 是一种独特的磷脂，只有在乙醇存在的情况下才会在体内形成。根据一项新的假设，血液中的高密度脂蛋白 (HDL) 颗粒可能充当 PEth 的载体，并介导适量饮酒的部分抗动脉粥样硬化作用。开发了使用反相 C8 柱和负离子模式电喷雾电离质谱检测和飞行时间 (TOF) 仪器的液相色谱方法，用于测定血液 HDL 颗粒上可能存在的极少量 PEth。当前研究中使用的样品是掺有 PEth 和内标磷脂酰丙醇 (PProp) 的人 HDL。反相色谱柱的使用使分析时间缩短为每次进样 19 分钟，仅为早期报道的正相方法的三分之一。由于色谱柱孔径窄（内径 2.1 分钟）和分析时间短，因此溶剂消耗量减少了。 TOF-MS 的检测灵敏度优于以前的方法，进样样品（20 pg ～ 28 fmol PEth）中的检测限低至 1 ng/ml，相当于每毫升未制备的 HDL 中含有约 6.7 ng PEth。在 1-100 ng/ml PEth 范围内获得了良好的检测线性，而精密度和准确度的所有偏差均小于 15%。 © 2005 Elsevier Inc. 保留所有权利。
Phosphatidylethanol (PEth) is a unique phospholipid that is formed in the body only in the presence of ethanol. According to a new hypothesis, blood high-density lipoprotein (HDL) particles may act as carriers of PEth and mediate part of the antiatherogenic effects of moderate alcohol drinking. Liquid chromatographic method using reversed-phase C8 column and negative ion mode electrospray ionization-mass spectrometry detection with time-of-flight (TOF) instrument was developed for the determination of very small amounts of PEth that might be present on blood HDL particles. The samples used in the current study were human HDL spiked with PEth and internal standard phosphatidylpropanol (PProp). The use of reversed-phase column enabled a short analysis time of 19 min/injection, which is only one-third of the earlier normal-phase methods reported. Because of the narrow bore column (2.1 min i.d.) and short analysis time, the solvent consumption was decreased. The sensitivity of detection obtained with TOF-MS was better than that of previous methods, with the detection limit being as low as 1 ng/ml in injected sample (20 pg on-column ∼ 28 fmol PEth), corresponding to approximately 6.7 ng of PEth in milliliter of unprepared HDL. Good linearity of detection was obtained for a range of 1-100 ng/ml of PEth, whereas all of the deviations in precision and accuracy were less than 15%. © 2005 Elsevier Inc. All rights reserved.