Faculty of Health Sciences, Medical Laboratory Sciences, Hokkaido University, Kita-12, Nishi-5, Sapporo, 060-0812, Japan.
Anal Bioanal Chem. 2011 Jun;400(7):1923-31. doi: 10.1007/s00216-011-4921-7. Epub 2011 Apr 2.
A new liquid chromatography-mass spectrometry (LC/MS) method has been developed for the quantitative analysis of plasmalogens in human plasma using a nonendogenous plasmalogen (1-O-1'-(Z)-tricosenyl-2-oleoyl-rac-glycero-3-phosphocholine, PLS 23:0/18:1) as an internal standard. 1-O-1'-(Z)-Tricosenyl glyceryl ether was prepared by reacting lithioalkoxyallyl with 1-iodoeicosane as the key intermediate in the formation of PLS 23:0/18:1. In LC/MS analyses, PLS 23:0/18:1 generated significant fragment ions in positive and negative modes. In positive ion mode, the M+H of PLS 23:0/18:1 yielded unique fragments with cleavages at the sn-1 and sn-2 positions of the glycerol backbone. In negative ion mode, the M+CH(3)COO of PLS 23:0/18:1 resulted in characteristic fragmentation at the sn-2 and sn-3 positions. 1-O-1'-(Z)-Hexadecenyl-2-linoleoyl-rac-glycero-3-phosphocholine (PLS 16:0/18:2) and 2-arachidonoyl-O-1'-(Z)-hexadecenyl-rac-glycero-3-phosphocholine (PLS 16:0/20:4) were chemically synthesized as PLS 23:0/18:1. The calibration curves obtained for PLS 16:0/18:2 and PLS 16:0/20:4 were linear throughout the calibration range (0.04-1.60 pmol). The LOD (S/N = 5:1) was 0.008 pmol and the LOQ (S/N = 6:1) was 0.01 pmol for both PLS 16:0/18:2 and PLS 16:0/20:4. Plasma concentrations of PLS 16:0/18:2 and PLS 16:0/20:4 were 4.0 ± 1.3 μM and 3.5 ± 1.2 μM (mean ± SD), respectively, in five healthy volunteers.
一种新的液相色谱-质谱（LC/MS）方法已经被开发出来，用于定量分析人血浆中的血浆类脂，使用非内源性的血浆类脂（1-O-1'-(Z)-三烯基-2-油酰基-rac-甘油-3-磷酸胆碱，PLS 23:0/18:1）作为内标。1-O-1'-(Z)-三烯基甘油醚是通过反应锂烷氧基烯丙基与 1-碘二十烷作为关键中间体来制备的，用于形成 PLS 23:0/18:1。在 LC/MS 分析中，PLS 23:0/18:1 在正离子和负离子模式下均产生显著的碎片离子。在正离子模式下，PLS 23:0/18:1 的M+H生成独特的片段，在甘油骨架的 sn-1 和 sn-2 位置发生断裂。在负离子模式下，PLS 23:0/18:1 的M+CH(3)COO在 sn-2 和 sn-3 位置产生特征的碎片。1-O-1'-(Z)-十六烯基-2-亚油酰基-rac-甘油-3-磷酸胆碱（PLS 16:0/18:2）和 2-花生四烯酸-O-1'-(Z)-十六烯基-rac-甘油-3-磷酸胆碱（PLS 16:0/20:4）被化学合成作为 PLS 23:0/18:1。PLS 16:0/18:2 和 PLS 16:0/20:4 的校准曲线在整个校准范围内呈线性（0.04-1.60 pmol）。PLS 16:0/18:2 和 PLS 16:0/20:4 的检出限（S/N = 5:1）为 0.008 pmol，定量限（S/N = 6:1）为 0.01 pmol。在五名健康志愿者中，PLS 16:0/18:2 和 PLS 16:0/20:4 的血浆浓度分别为 4.0 ± 1.3 μM 和 3.5 ± 1.2 μM（平均值±SD）。
