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.
Se Pu. 2018 Oct 8;36(10):985-990. doi: 10.3724/SP.J.1123.2018.05029.
A method for the determination of cholesterol and its six metabolism markers (desmosterol, lathosterol, campesterol, stigmasterol, -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 C 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 MS (Full MS/dd MS) mode. The resolution of the precursor mass was 70000 FWHM, while the resolution of the product mass was 17500 FWHM. The correlation coefficients () 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.
建立了一种采用高效液相色谱-四极杆/静电场轨道阱高分辨质谱（HPLC-Q/Orbitrap HRMS）测定血清中胆固醇及其六种代谢标志物（羊毛甾醇、胆甾烷醇、菜油甾醇、豆甾醇、β-谷甾醇、角鲨烯）的方法。样品用乙腈超声提取。以95%（v/v）甲醇-乙腈（2:8，v/v）-5%（v/v）水为流动相进行分离。在Acquity UPLC BEH C柱（100 mm×2.1 mm，1.7 μm）上以0.4 mL/min的流速进行等度洗脱，洗脱时间为15 min。质谱检测采用大气压化学电离（APCI）/正模式。采用全扫描质谱/数据依赖型质谱（Full MS/dd MS）模式对胆固醇及其六种代谢标志物进行定性和定量测定。母离子质量分辨率为70000 FWHM（半高宽），子离子质量分辨率为17500 FWHM。七种目标化合物在其线性范围内的相关系数均不小于0.992。检测限在0.8 μg/L至62.1 μg/L之间，回收率在82.1%至97.5%之间，相对标准偏差在1.6%至7.4%之间。该方法准确、简便、快速，可用于血清中胆固醇及其六种代谢标志物的测定。
