Mi Si, Zhao Yuan-Yuan, Dielschneider Rebecca F, Gibson Spencer B, Curtis Jonathan M
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada.
Department of Immunology, University of Manitoba, Winnipeg, Manitoba, Canada; Research Institute of Oncology and Hematology, Cancer Care Manitoba, Winnipeg, Manitoba, Canada.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Sep 15;1031:50-60. doi: 10.1016/j.jchromb.2016.07.028. Epub 2016 Jul 16.
Comprehensive profiling of sphingolipids is of great importance for clinical and pharmaceutical studies. An LC/MS/MS method was established for the simultaneous separation and quantification of individual sphingolipid species including ceramides, dihydroceramides, glucosylceramides, sphingosine, sphingosine-1-phosphate, sphinganine and sphinganine-1-phosphate. All target individual sphingolipid species were separated and quantified in a single chromatographic run of <20min. Method validation results indicated that calibration curves were linear in the range of 2.5-10,000nM for ceramides and glucosylceramides, 10-10,000nM for dihydroceramides, 5-10,000nM for sphingosine, sphingosine-1-phosphate, sphinganine and sphinganine-1-phosphate, respectively. The limits of detection ranged from 0.5nM to 5nM. Accuracies of 92.5-113% with precisions of 0.3-8.0% RSD were obtained for all of the standards over a wide range of concentrations. The application of this method was demonstrated using B cells collected from Chronic Lymphocytic Leukemia patients (n=5) and healthy donors (n=4). 17 sphingolipid species were successfully characterized and quantified in the lipid extract. This is a rapid method that could be readily adapted to lipidomic investigations of sphingolipids in other bio-fluids and tissues.
鞘脂的全面分析对于临床和药学研究具有重要意义。建立了一种液相色谱/串联质谱法，用于同时分离和定量分析包括神经酰胺、二氢神经酰胺、葡萄糖神经酰胺、鞘氨醇、鞘氨醇-1-磷酸、二氢鞘氨醇和二氢鞘氨醇-1-磷酸在内的各鞘脂种类。所有目标鞘脂种类在不到20分钟的单次色谱运行中即可分离和定量。方法验证结果表明，神经酰胺和葡萄糖神经酰胺的校准曲线在2.5 - 10,000 nM范围内呈线性，二氢神经酰胺在10 - 10,000 nM范围内，鞘氨醇、鞘氨醇-1-磷酸、二氢鞘氨醇和二氢鞘氨醇-1-磷酸分别在5 - 10,000 nM范围内。检测限为0.5 nM至5 nM。在广泛的浓度范围内，所有标准品的准确度为92.5 - 113%，精密度为0.3 - 8.0% RSD。使用从慢性淋巴细胞白血病患者（n = 5）和健康供体（n = 4）收集的B细胞证明了该方法的应用。在脂质提取物中成功鉴定并定量了17种鞘脂种类。这是一种快速方法，可很容易地应用于其他生物流体和组织中鞘脂的脂质组学研究。
