Lithium Hydroxide Hydrolysis Combined with MALDI TOF Mass Spectrometry for Rapid Sphingolipid Detection.
Lithium Hydroxide Hydrolysis Combined with MALDI TOF Mass Spectrometry for Rapid Sphingolipid Detection.
复制标题
DOI：
10.1021/jasms.0c00322
复制
发表时间：
2021-01-06
影响因子：
3.2
通讯作者：
Jones JW
中科院分区：
文献类型：
--
作者：
Tran A;Wan L;Xu Z;Haro JM;Li B;Jones JW
鞘脂具有多样的结构和生物活性功能，在许多关键的生物学过程中发挥重要作用。相对含量低、结构多样以及浓度动态范围等因素为鞘脂检测带来了困难的分析挑战。为了进一步改进基于质谱的鞘脂分析，实施了锂加合物整合以降低光谱复杂性并合并信号强度，从而提高了特异性和灵敏度。我们报道在常规水解过程中使用氢氧化锂作为碱，以便有效去除常见的电离抑制剂（如糖脂和甘油磷脂）并向样品中引入锂源。同时，使用一种优化的基质辅助激光解吸/电离（MALDI）基质系统，其以2′,4′,6′ - 三羟基苯乙酮（THAP）为特色，用于在MALDI过程中促进锂加合物整合。其结果是一种稳健且高通量的鞘脂检测方案，特别是针对低丰度的神经酰胺。我们所开发的工作流程的应用包括检测高脂肪诱导的肥胖小鼠模型中肝脏鞘脂谱的差异表达。我们还证明了该方法在检测大脑和血浆基质中各种鞘脂的有效性。这些结果通过超高效液相色谱 - 高分辨质谱/质谱（UHPLC HR MS/MS）和基质辅助激光解吸/电离 - 傅里叶变换离子回旋共振（MALDI FT - ICR）的数据得到了证实，验证了该方法应用的有效性。总体而言，我们展示了一种通过使用基质辅助激光解吸/电离飞行时间质谱（MALDI TOF）和锂加合物整合在各种生物基质中进行鞘脂分析的高通量工作流程。
Sphingolipids have diverse structural and bioactive functions that play important roles in many key biological processes. Factors such as low relative abundance, varied structures, and a dynamic concentration range provide a difficult analytical challenge for sphingolipid detection. To further improve mass-spectrometry-based sphingolipid analysis, lithium adduct consolidation was implemented to decrease spectral complexity and combine signal intensities, leading to increased specificity and sensitivity. We report the use of lithium hydroxide as a base in a routine hydrolysis procedure in order to effectively remove common ionization suppressants (such as glycolipids and glycerophospholipids) and introduce a source of lithium into the sample. In conjunction, an optimized MALDI matrix system, featuring 2′,4′,6′-trihydroxyacetophenone (THAP) is used to facilitate lithium adduct consolidation during the MALDI process. The result is a robust and high-throughput sphingolipid detection scheme, particularly of low-abundance ceramides. Application of our developed workflow includes the detection of differentially expressed liver sphingolipid profiles from a high-fat-induced obesity mouse model. We also demonstrate the method’s effectiveness in detecting various sphingolipids in brain and plasma matrices. These results were corroborated with data from UHPLC HR MS/MS and MALDI FT-ICR, verifying the efficacy of the method application. Overall, we demonstrate a high-throughput workflow for sphingolipid analysis in various biological matrices by the use of MALDI TOF and lithium adduct consolidation.
登录
查看更多内容
DOI：
10.1016/j.jchromb.2008.12.057
发表时间：
2009-09-15
期刊：
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
影响因子：
--
作者：
Haynes CA;Allegood JC;Park H;Sullards MC
通讯作者：
Sullards MC
DOI：
10.1038/nrm.2017.107
发表时间：
2018-03
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Hannun YA;Obeid LM
通讯作者：
Obeid LM
影响因子：
2.9
作者：
Jiang, Xuntian;Cheng, Hua;Han, Xianlin
通讯作者：
Han, Xianlin
DOI：
10.1016/j.jchromb.2008.05.013
发表时间：
2008-07-15
影响因子：
3
作者：
Fujiwaki, Takehisa;Tasaka, Masaru;Yamaguchi, Seiji
通讯作者：
Yamaguchi, Seiji
影响因子：
2
作者：
Griffiths, Rian L.;Bunch, Josephine
通讯作者：
Bunch, Josephine