Comprehensive untargeted lipidomic analysis using core-shell C30 particle column and high field orbitrap mass spectrometer.
Comprehensive untargeted lipidomic analysis using core-shell C30 particle column and high field orbitrap mass spectrometer.
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DOI：
10.1016/j.chroma.2016.02.054
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发表时间：
2016-04-01
期刊：
影响因子：
--
通讯作者：
Zhang Q
中科院分区：
文献类型：
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作者：
Narváez-Rivas M;Zhang Q
非靶向脂质组学的目标是高通量、全面和明确地识别和定量脂类。为了实现这一目标，新的LC分离固定相和新的具有高质量分辨率和更快扫描速度的质谱仪是必不可少的。用4根反相色谱柱与高场四极轨道质谱仪(Q Exactive HF)联用，用复杂的脂质标准混合物和大鼠血浆和肝脏样品进行了全面的比较。在24分钟的梯度条件下，所有的脂类都得到了很好的分离。比较的柱包括C30和C18在核壳或全孔二氧化硅颗粒上的官能化，尺寸在1.7~2.6μm之间。Accucore C30柱具有最窄的峰和最高的理论板数，良好的峰容量和保留时间重现性(<；1%标准偏差)。结果，它以最高的置信度从大鼠血浆和大鼠肝脏样本中鉴定出430种脂类。RPLC提供的高分辨率可以区分顺/反异构体的脂类，而高场Orbitrap质谱仪在全扫描MS中可以清楚地区分等压脂类，在MS/MS中可以明确地指定溶血磷脂的sn位置异构体。
The goal of untargeted lipidomics is to have high throughput, yet comprehensive and unambiguous identification and quantification of lipids. Novel stationary phases in LC separation and new mass spectrometric instruments capable of high mass resolving power and faster scanning rate are essential to achieving this goal. In this work, 4 reversed phase LC columns coupled with a high field quadrupole orbitrap mass spectrometer (Q Exactive HF) were thoroughly compared using complex lipid standard mixture and rat plasma and liver samples. A good separation of all lipids was achieved in 24 min of gradient. The columns compared include C30 and C18 functionalization on either core–shell or totally porous silica particles, with size ranging from 1.7 to 2.6 μm. Accucore C30 column showed the narrowest peaks and highest theoretical plate number, and excellent peak capacity and retention time reproducibility (<1% standard deviation). As a result, it resulted in 430 lipid species identified from rat plasma and rat liver samples with highest confidence. The high resolution offered by the up-front RPLC allowed discrimination of cis/trans isomeric lipid species, and the high field orbitrap mass spectrometer afforded the clear distinction of isobaric lipid species in full scan MS and the unambiguous assignment of sn-positional isomers for lysophospholipids in MS/MS. Taken together, the high efficiency LC separation and high mass resolving MS analysis are very promising tools for untargeted lipidomics analysis.