以 LC-ESI-MS 分析三種不同疾病其腦脊液中 glycerophospholipids 和 sphingolipid 的特性   全文下載

Characterization of Glycerophospholipids and a Sphingolipid in the Cerebrospinal Fluid of Patients with Different Illnesses by LC-ESI-MS

一個用來測量glycerophospholipids和sphingolipid的靈敏且準確的逆相液相層析連結電灑游離質譜儀（LC-ESI-MS）分析方法已被發展和確效；並且應用在三種不同疾病的病人其腦脊液中脂質的分析。液相層析電灑游離法質譜儀分析glycerophospholipids和sphingolipid的最適宜條件為使用Inertsil 6 ODS-3（4.6 × 150 mm）層析管柱，移動相包含了乙?／甲醇／三乙胺，比例分別為550/1000/25（w/w/w），固定流速為1.0 mL/min。結果證實phosphatidylcholine（PC）、phospatidylethanolamine（PE）、phosphatidylino-sitol（PI）、phosphatidylsetine（PS）以及sphingomyelin（SM）在同日間及異日間的準確度及精密度皆在可接受的範圍。臨床應用顯示在三種不同疾病其腦脊液的種類及含量比例皆不相同。在背脊上皮瘤（chordoma）、三叉神經痛（trigeminal neuralgia）以及帕金森氏症（Parkinson’s disease）三種病人其脂質分別以SM（848 ± 75.5 ng/mL, 40.7% ± 2.2%）、PC（8042.5 ± 1214.4 ng/mL, 53.8% ± 4.2%）及PS（2124.2 ± 152.0 ng/mL, 43.3% ± 1.6%）含量最多。總而言之，此分析方法可以直接檢測glycerophospholipids和sphingolipid，並且應用到偵測在人體上的生理變化以發展預防性或治療性醫學以及新藥發展。"

A sensitive and accurate reversed-phase liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MS) method for determining glycerophospholipids and a sphingolipid was developed and validated in order to profile and quantify the classes of lipids in the cerebrospinal fluids of patients with three different illnesses. The LC-ESI-MS method was optimized for the analysis, and an Inertsil 6 ODS-3 (4.6 × 150 mm) column was utilized with a mobile phase composed of acetonitrile/methanol/triethyl-amine in the ratio 550/1000/25 (w/w/w) eluted isocratically at a flow rate of 1.0 mL/min. The results demonstrated that both the accuracy and precision of the intra- and inter-day assays of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and sphingomyelin (SM) were within acceptable criteria. Clinical application demonstrated that the percentage and content of each class differed in the cerebrospinal fluids of patients with three different illnesses, with the richest lipid components being SM (848 ± 75.5 ng/mL, 40.7% ± 2.2%), PC (8042.5 ± 1214.4 ng/mL, 53.8% ± 4.2%) and PS (2124.2 ± 152.0 ng/mL, 43.3% ± 1.6%), respectively for patients with chordoma, trigeminal neuralgia and Parkinson’s disease. It was concluded that this improved method can be used to directly detect glycerophospholipids and a sphingolipid, and applied to detect physiological changes in humans to discover preventive medicines or therapeutic medicines, or develop new drugs.