固相萃取-超高效液相色谱-静电场轨道阱高分辨率质谱法测定血浆中神经酰胺  

Determination of Ceramides in Plasma by Solid Phase Extraction-Ultra High Performance Liquid Chromatography-Tandem Orbitrap High Resolution Mass Spectrometry

作　　者：吴思非 张雅 刘英杰 孙玉洁 张晓光[3] WU Sifei;ZHANG Ya;LIU Yingjie;SUN Yujie;ZHANG Xiaoguang(School of Parmacy,Hebei Medical University,Shijiazhuang 050017;School of Nursing,Hebei Medical University,Shijiazhuang 050035;Core Facilities and Centers,Hebei Medical University,Shijiazhuang 050017)

机构地区：[1]河北医科大学药学院,河北石家庄050017 [2]河北医科大学护理学院,河北石家庄050035 [3]河北医科大学大型科研仪器设备共享服务平台,河北石家庄050017

出　　处：《分析科学学报》2024年第6期689-695,共7页Journal of Analytical Science

基　　金：河北省高等学校科学技术研究项目(产学研合作)(CXY2024051);河北省卫生健康委员会医学科学研究课题项目(20200038)。

摘　　要：建立了固相萃取-超高效液相色谱-静电场轨道阱高分辨率质谱测定血浆中10种神经酰胺的方法。样品经甲醇提取、固相萃取柱净化,采用C30色谱柱进行色谱分离;以母离子精确质量数提取峰面积进行定量,以相对应的子离子进行定性,用同位素内标法建立标准曲线。实验对固相萃取柱、色谱柱,以及固相萃取洗脱条件、色谱条件以及质谱条件进行优化,并考察了不同净化方法的基质效应。10种神经酰胺在一定的线性范围内拟合良好(相关系数r^(2)>0.99),检出限范围为0.5~1.2 ng/mL,定量限范围为1.5~3.5 ng/mL。方法的回收率在73.1%~106.9%之间,相对标准偏差在1.0%~13.1%之间。方法操作方便,分析时间短,准确性高,可为神经酰胺的测定以及相关研究提供新的技术支持。A method was established for the simultaneous detection of 10 ceramides in serum by solid phase extraction-ultra high performance liquid chromatography-orbitrap high resolution mass spectrometry.The sample was purified by a solid phase extraction column and separated by C30 chromatographic column after methanol extraction.The peak area was extracted by the exact mass of the parent ion for quantification,and the corresponding product ion for qualitative analysis.The standard curve was established by an isotope internal standard method.The solid phase extraction column and chromatographic column were selected,and the elution conditions of solid phase extraction,chromatographic conditions and mass spectrometry conditions were optimized.The matrix effects of different purification methods were compared.The 10 analytes showed good linearity within the specified range(r^(2)>0.99).The limit of detection(LOD)ranged from 0.5 ng/mL to 1.2 ng/mL while the limit of quantification(LOQ)ranged from 1.5 ng/mL to 3.5 ng/mL.The recovery ranged from 73.1% to 106.9%,and the relative standard deviation(RSD)ranged from 1.0% to 13.1%.This method has the advantages of easy operation,short operation time and high accuracy,which provides a novel technical support for the determination of ceramide and related research.

关 键 词：血浆 神经酰胺 固相萃取 高分辨率质谱 

分 类 号：O657.63[理学—分析化学]