中文摘要：

目的建立并确证可以检测1-磷酸鞘氨醇（S1P）快速、灵敏、特异的LC—MS／MS定量分析方法。方法采用C17-SIP作为内参，甲醇一步法进行蛋白沉淀，随后进行正相电喷雾离子化LC—MS／MS分析。流动相为甲醇-a％甲酸水（95：5，v／v），流速0．2mL／min，每个样品的分析时间为4min。细胞经TNF—a处理后SIP含量显著增加；而经DMs处理后S1P含量显著下降。结果SIP的标准曲线线性范围为0．1。10ng／mL。相关系数r2均大于0．989。结论该方法可以快速，灵敏，特异性地同时检测生物样品中SIP的含量。

英文摘要：

Objective In the present study,we developed a rapid, sensitive and specific LC-MS/MS method to determine the levels of S1P in biological samples. Methods C17-S1P were used as internal standards. With one step of methanol-induced protein precipitation, each sample was subjected to LC-MS/MS analysis using positive electrospray ionization under selected reaction monitoring mode. The running time was within 4 rain with a simple mobile phase consisting of methanol-0.1% formic acid （95 ： 5,v/v） at a flow rate of 0.2 mL/min. The SIP levels increased in cells treated with TNF- a whereas decreased in cells treated with DMS. Results Standard curves were linear over ranges of 0.1 - 10 ng/mL for S1P with r2 greater than 0.989. Conclusion These results indicated that this optimized LC-MS/MS method was rapid, sensitive, specific and reliable to quantify S1P levels in biological samples.