Off-line mixed-mode liquid chromatography coupled with reversed phase high performance liquid chromatography-high resolution mass spectrometry to improve coverage in lipidomics analysis.
Off-line mixed-mode liquid chromatography coupled with reversed phase high performance liquid chromatography-high resolution mass spectrometry to improve coverage in lipidomics analysis.
复制标题
离线混合模式液相色谱与反向相位高性能液相色谱高分辨率质谱法相反，以改善脂质分析中的覆盖率。
DOI：
10.1016/j.aca.2016.12.003
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发表时间：
2017-02-15
影响因子：
6.2
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Narváez-Rivas M;Vu N;Chen GY;Zhang Q
可靠的识别和深入分析分子脂质种类仍然是脂质组学分析中的一个挑战。本研究建立了一种离线二维混合模式反相液相色谱（RPLC）与高场四极杆轨道阱质谱仪（Q Exactive HF）联用的方法，用于复杂生物样品中脂质的分析。在第一维中，22种不同的脂质类在整体硅胶柱上分离，洗脱顺序从中性到极性脂质。收集总共13个级分，并在RPLC C30柱上以第二维运行，以基于其疏水性进一步分离脂质分子种类，洗脱顺序由脂肪酰基链中的长度和不饱和度确定。应用该方法对大鼠血浆和肝脏中的脂质进行了分析。通过气相色谱-质谱法进行脂肪酸甲酯分析，以确定总脂质提取物中的脂肪酰基，这提供了一个更可靠的识别这些样品中存在的脂质种类。在每个样品中鉴定了800多种脂质，并使用串联质谱法（MS/MS）机密地确认了它们的分子结构。通过这种离线二维方法在大鼠血浆和大鼠肝脏中鉴定的脂质分子种类的数量约为采用C30柱的一维RPLC-MS/MS的两倍。这种离线二维混合模式LC-RPLC-MS/MS方法是一种很有前途的技术，用于复杂生物基质中的全面脂质分析。
The confident identification and in-depth profiling of molecular lipid species remain to be a challenge in lipidomics analysis. In this work, an off-line two-dimensional mixed-mode and reversed-phase liquid chromatography (RPLC) method combined with high-field quadrupole orbitrap mass spectrometer (Q Exactive HF) was developed to profile lipids from complex biological samples. In the first dimension, 22 different lipid classes were separated on a monolithic silica column with elution order from neutral to polar lipids. A total of 13 fractions were collected and run on a RPLC C30 column in the second dimension for further separation of the lipid molecular species based on their hydrophobicity, with the elution order being determined by both the length and degree of unsaturation in the fatty-acyl chain. The method was applied to analyze lipids extracted from rat plasma and rat liver. Fatty acid methyl ester analysis by gas chromatography-mass spectrometry was used to identify the fatty acyls from total lipid extracts, which provided a more confident identification of the lipid species present in these samples. More than 800 lipids were identified in each sample and their molecular structures were confidentially confirmed using tandem mass spectrometry (MS/MS). The number of lipid molecular species identified in both rat plasma and rat liver by this off-line two-dimensional method is approximately twice of that by one-dimensional RPLC-MS/MS employing a C30 column. This off-line two-dimensional mixed-mode LC-RPLC-MS/MS method is a promising technique for comprehensive lipid profiling in complex biological matrices.