Lithium Hydroxide Hydrolysis Combined with MALDI TOF Mass Spectrometry for Rapid Sphingolipid Detection.
Anh TranLiting WanZhenbo XuJanette M HaroBing LiJace W JonesPublished in: Journal of the American Society for Mass Spectrometry (2020)
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.
Keyphrases
- mass spectrometry
- liquid chromatography
- loop mediated isothermal amplification
- gas chromatography
- ms ms
- high throughput
- high performance liquid chromatography
- solid state
- high resolution
- capillary electrophoresis
- high resolution mass spectrometry
- mouse model
- label free
- real time pcr
- tandem mass spectrometry
- randomized controlled trial
- type diabetes
- metabolic syndrome
- magnetic resonance
- systematic review
- insulin resistance
- computed tomography
- body mass index
- ultra high performance liquid chromatography
- physical activity
- clinical practice
- liquid chromatography tandem mass spectrometry
- skeletal muscle
- sensitive detection
- adipose tissue
- subarachnoid hemorrhage
- white matter
- artificial intelligence