Covalent Organic Framework Nanofilm-Based Laser Desorption/Ionization Mass Spectrometry for 5-Fluorouracil Analysis and Tissue Imaging

Although matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has become a ubiquitous and effective tool for macromolecules, direct analysis of small molecules by MALDI-MS using conventional organic matrices poses a challenge. Herein, a large-area, uniform, and stable covalent organic framework (COF) nanofilm prepared directly on indium-tin oxide (ITO) glass was first introduced as a substrate for LDI-MS, which showed enhanced sensitivity, no background interference, and high reproducibility in the analysis of diverse small molecules. Taking into account all these merits, an attractive approach of COF nanofilm-based LDI-MS was developed to quantitatively evaluate the pharmacokinetics of 5-fluorouracil (5-FU) in mouse plasma. A good linear relationship (10-20,000 ng/mL) and a low limit of detection (LOD) for 5-FU (∼100 pg/mL) were achieved. In view of the fact that the COF nanofilm was uniform and without the requirement of additional matrix spraying, it was further extended for LDI-MS imaging (LDI-MSI) to visualize the spatial distribution of 5-FU in mouse liver at different interval times after intravenous and intragastric administrations. The results indicated that the decay of 5-FU in mouse liver obtained with the COF nanofilm-based LDI-MSI was consistent with the tendency of 5-FU pharmacokinetics. This work not only offers an alternative solution for LDI-MS/MSI analysis of small molecules but also extends the application fields of COF nanofilm in MS research.