A rapid evaporative ionization mass spectrometry (REIMS) method was developed for lipidomics tracking of Ctenopharyngodon idellus during in vitro multiple-stage digestion. The REIMS conditions were optimized such that the temperature of the heating probe was 500 °C, sample amount was 30 mg, and the flow rate of auxiliary solvent was 100 μL min. The results showed that the phospholipids were detected with variety and quantity in the crude and multiple-stage digested samples. The enzymatic effect on the phospholipids is varied depending on the phospholipid classes, and the hydrolysis rate of phospholipids increased as the degree of unsaturation of the acyl chain increased. The principal component analysis (PCA) indicated that the ions at m/ z 809.61, 811.63, and 857.52 were the most noticeable species digested during the process. This method exhibited great potential in fast lipidomics profiling for inspecting the characteristics of nutritional lipid absorption digestion in human gastrointestin.
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