高效液相色谱-四极杆/静电场轨道阱高分辨质谱法测定血清中胆固醇及其6种代谢标志物  

Determination of cholesterol and its six metabolism markers in serum by high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry

作　　者：黄雯雯[1] 勾新磊[1] 胡铁靖 胡光辉[1] 赵新颖[1] HUANG Wenwen;GOU Xinlei;HU Tiejing;HU Guanghui;ZHAO Xinying(Beijing Center for Physical and Chemical Analysis,Beijing Key Laboratory of Organic Materials Testing Technology ＆ Quality Evaluation,Beijing 100094,China)

机构地区：[1]北京市理化分析测试中心有机材料检测技术与质量评价北京市重点实验室,北京100094

出　　处：《色谱》2018年第10期985-990,共6页Chinese Journal of Chromatography

基　　金：北京市科学技术研究院萌芽计划(2017-01);北京市自然科学基金-北京市科学技术研究院联合资助项目(L160005)~~

摘　　要：利用高效液相色谱-四极杆/静电场轨道阱高分辨质谱(HPLC-Q/OrbitrapHRMS)测定血清中的胆固醇及其6种代谢标志物(2,4-脱氢胆甾醇、7-烯胆甾醇、菜油固醇、豆固醇、β-谷固醇和角鲨烯)。以乙腈作为提取溶剂,超声提取,采用Acquity UPLC BEH C18色谱柱(100 mm×2. 1 mm,1. 7μm),以95%(v/v)的甲醇-乙腈(2∶8,v/v)-5%(v/v)水做流动相,等度洗脱15 min,流速0. 4 mL/min。在大气压化学电离源(APCI)正离子扫描模式下,质谱采用全扫描/数据依赖二级扫描(Full MS/dd MS2)监测模式,同时获得定性和定量结果,一级分辨率为70 000FWHM,二级分辨率为17 500 FWHM。7种目标物质在其线性范围内的线性相关系数(r2)均不小于0. 992,检出限为0. 8~62. 1μg/L,3个加标水平下的回收率为82. 1%~97. 5%,相对标准偏差(RSD)为1. 6%~7. 4%。方法准确、简单、快捷,可以作为血清中胆固醇及其6种代谢标志物的检测方法。A method for the determination of cholesterol and its six metabolism markers （desmosterol, lathosterol, campesterol, stigmasterol, β-sitosterol, squalene） in serum by high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （HPLC-Q/Orbitrap HRMS） was developed. The sample was ultrasonically extracted with acetonitrile. The separation was performed by using 95%（v/v） methanol-acetonitrile （2：8, v/v）-5%（v/v） water as the mobile phase. Isocratic elution was performed at a flow rate of 0.4 mL/min on an Acquity UPLC BEH C18 column （100 mm×2.1 mm, 1.7 μm） in 15 min. The atmospheric pressure chemical ionization （APCI）/positive mode was used in the MS detection. Cholesterol and its six metabolism markers were qualitatively and quantitatively determined by using the full MS scan/date dependent MS2 （Full MS/dd MS2） mode. The resolution of the precursor mass was 70000 FWHM, while the resolution of the product mass was 17500 FWHM. The correlation coefficients （r2） of all the seven target compounds were no less than 0.992 in the linear range. The limits of detection were between 0.8 μg/L and 62.1 μg/L, the recoveries were in the range of 82.1%-97.5% with the relative standard deviations in range of 1.6%-7.4%. The method is accurate, simple, and rapid, and can be used for the determination of cholesterol and its six metabolism markers in serum.

关 键 词：高效液相色谱 四极杆/静电场轨道阱高分辨质谱 胆固醇 胆固醇代谢标志物 血清 

分 类 号：O658[理学—分析化学]