检测细胞中1-磷酸鞘氨醇含量的LC-MS/MS分析方法的建立  

Development of aLC-MS/MS method for determination ofsphingosine 1-phosphate levels in biological samples

作　　者：兰天[1] 吴腾[1] 勾红菊[1] 吴平香[1] 常秀亭[2] 王丽京[1] 黄河清[2] 

机构地区：[1]广东药学院血管生物学研究所,广东广州510006 [2]中山大学药学院,广东广州510006

出　　处：《中国医药科学》2013年第9期9-11,共3页China Medicine And Pharmacy

基　　金：国家自然科学基金(81170676;81200308;31271455);广东省自然科学基金(S2012020010991)

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-S1P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-0.1%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-α处理后S1P含量显著增加;而经DMS处理后S1P含量显著下降。结果 S1P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S1P的含量。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.

关 键 词：含量测定 1-磷酸鞘氨醇 液质联用 

分 类 号：R735.205[医药卫生—肿瘤]