液-质联用法对生物样本中鞘氨醇1-磷酸的微量测定  

Microdetermination of Sphingosine 1-phosphate from Biological Samples by Liquid Chromatography Mass Spectrometry

作　　者：郭守东 于杨[2] 冯蕾[1] 刘帅[1,2] 李和楼[3] 崔英杰[1] 秦树存[1,2] 

机构地区：[1]山东省高校动脉粥样硬化重点实验室脂质分析实验室,泰山医学院动脉粥样硬化研究所 [2]山东省高校动脉粥样硬化重点实验室基因功能实验室,泰山医学院动脉粥样硬化研究所 [3]山东省泰山医学院附属医院,山东省泰安市271000

出　　处：《中国动脉硬化杂志》2013年第4期354-358,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(30971098;81070247);山东省泰山学者岗专项基金(ZD056;ZD057);2012年泰山医学院高层次课题培养计划项目(330033)

摘　　要：目的本文旨在建立一种简便的鞘氨醇1-磷酸分离提取方法,并结合液-质联用法快速检测生物样本中纳克水平的鞘氨醇1-磷酸。方法采用甲醇沉淀结合超声萃取的方法分离鞘氨醇1-磷酸,高速离心去除大颗粒后的上清液采用液-质联用仪在选择性离子监测模式下进行定量分析。结果该分离提取鞘氨醇1-磷酸的方法简便快捷,鞘氨醇1-磷酸在选择性离子监测模式下的定量限为159.7 pg,其在人血浆中的含量为128.8±33.3μg/L;在C57 BL/6小鼠血浆、心、肝、肺、肾和脑中的含量分别为181.8±21.1μg/L、37.6±2.8 ng/g、54.9±4.9 ng/g、230.1±60.5 ng/g、21.5±6.7 ng/g、102.2±23.8 ng/g。结论甲醇提取分离法可简便快捷地获得血浆及脏器中的鞘氨醇1-磷酸,结合液-质联用仪可对血浆、组织脏器中的鞘氨醇1-磷酸进行定量检测具有高效、重现性好等优点,特别适用于大量生物样本的快速处理与分析。Aim To establish a simplified extraction method from biological sample, and quantify sphingosine 1- phosphate （SIP） at the level of nanogram. Methods The methanol precipitation combing with ultrasonic extraction method was used to extract and separate S1 P, then the supernatant was analyzed by liquid chromatography mass spectrome- try （LC-MS） under the selective ion monitoring mode. Results The extraction method was effective and time saving, the limitation of quantification of S1P was 159. 7 pg; The S1P content in human plasma was 128. 8 ＋ 33.3μg/L, and the SIP content in plasma, heart, liver, lung, kidney and brain of the C57 BL/6 mouse was 181.8 ± 21.1 μg/L, 37. 6± 2. 8 ng/g, 54.9±4. 9 ng,/g, 230. 1±60. 5 ng./g, 21.5 ±6. 7 ng/g and 102. 2± 23.8 ng/g, respectively. Conelusion Methanol precipitation combing with ultrasonic extraction could effectively separate S1P from plasma and organ, and the extract could be quantified by LC-MS directly. This method is effective with good reproducibility, and is suitable for quick quantitative analysis of a great deal of biological sample.

关 键 词：鞘氨醇1-磷酸 液-质联用仪 生物样本 

分 类 号：R313[医药卫生—基础医学]