Two-dimensional liquid chromatography-mass spectrometry for lipidomics using off-line coupling of hydrophilic interaction liquid chromatography with 50 cm long reversed phase capillary columns
Two-dimensional liquid chromatography-mass spectrometry for lipidomics using off-line coupling of hydrophilic interaction liquid chromatography with 50 cm long reversed phase capillary columns
复制标题
使用亲水相互作用液相色谱与 50 cm 长反相毛细管柱的离线耦合进行脂质组学二维液相色谱-质谱分析
DOI：
10.1016/j.chroma.2022.463707
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发表时间：
2023
影响因子：
4.1
通讯作者：
Kennedy, Robert T.
中科院分区：
文献类型：
--
作者：
Sorensen, Matthew J.;Miller, Kelsey E.;Jorgenson, James W.;Kennedy, Robert T.
脂质组的全面表征仍然是一个挑战，需要开发新的分析方法来扩大复杂样品中的脂质覆盖范围。本研究采用离线二维液相色谱-质谱联用技术对人血浆中的脂质组学进行了研究。亲水相互作用液相色谱法在第一维实施，以分离脂质类。收集9个馏分，并使用填充有1.7 µm C18颗粒的50 cm毛细管柱在35 kpsi的定制仪器上进行二维分离。在线耦合飞行时间质谱法允许推定的脂质识别从基于质谱的库搜索。该方法具有良好的正交性（分数覆盖率≥ 40%），在600 min内达到约1900的峰容量，并从5 µL人血浆提取物进样中检测到超过1000种脂质，同时消耗不到3 mL溶剂。结果表明，采用长柱和二维分离时，峰容量的预期收益，并说明了实用的方法，用于改善复杂的生物样品的脂质体覆盖。
Comprehensive characterization of the lipidome remains a challenge requiring development of new analytical approaches to expand lipid coverage in complex samples. In this work, offline two-dimensional liquid chromatography-mass spectrometry was investigated for lipidomics from human plasma. Hydrophilic interaction liquid chromatography was implemented in the first dimension to fractionate lipid classes. Nine fractions were collected and subjected to a second-dimension separation utilizing 50 cm capillary columns packed with 1.7 µm C18 particles operated on custom-built instrumentation at 35 kpsi. Online coupling with time-of-flight mass spectrometry allowed putative lipid identification from precursor-mass based library searching. The method had good orthogonality (fractional coverage of ∼40%), achieved a peak capacity of approximately 1900 in 600 min, and detected over 1000 lipids from a 5 µL injection of a human plasma extract while consuming less than 3 mL of solvent. The results demonstrate the expected gains in peak capacity when employing long columns and two-dimensional separations and illustrate practical approaches for improving lipidome coverage from complex biological samples.