Enhanced imaging of endogenous metabolites by negative ammonia assisted DESI/PI mass spectrometry.

In this work, endogenous metabolites in mouse brain tissue were imaged by negative desorption electrospray ionization (DESI), ammonia assisted DESI (aa-DESI), DESI/post-photoionization (DESI/PI), and ammonia assisted DESI/PI (aa-DESI/PI) mass spectrometry imaging (MSI) strategies. The combined effect of ammonia additive and post-photoionization was found to play an important role in the enhancement of sensitivity and coverage for endogenous analytes under ambient conditions. Compared with DESI, aa-DESI/PI can provide increased signal intensities for metabolites up to 37.1-fold, as well as the imaging of nine more small metabolites (m/z < 350) (26 for aa-DESI/PI and 17 for DESI) in mouse brain tissue. The results of Pearson correlation analysis and KEGG pathway analysis showed that the enhanced imaging strategy of aa-DESI/PI can facilitate the study of endogenous metabolic pathways and molecular networks. Moreover, the imaging results of mouse tumor tissue demonstrated the promising application of aa-DESI/PI in tumor research.

Enhanced Coverage and Sensitivity of Imprint DESI Mass Spectrometry Imaging for Plant Leaf Metabolites by Post-photoionization.

Plant metabolites exhibit a variety of different chemical properties, physiological activities, and biological functions. However, untargeted imaging of highly diverse metabolic profiles is still a great challenge. Here, metabolites in plant leaves were imaged via imprint, followed by desorption electrospray ionization/post-photoionization (imprint DESI/PI) mass spectrometry imaging. In contrast to the traditional imprint DESI method, quite a few metabolites, such as terpenoids, flavonoids, glycosides, alkylphenols, amino acids, phenolic acids, tannins, and lipids, in fresh sage leaves, ginkgo leaves, and tea leaves were well detected and imaged by imprint DESI/PI. More than 80 metabolites were additionally identified, and more than 1 order of magnitude higher signal intensities were obtained for most metabolites in the negative ion mode. By virtue of the significant improvement of coverage and sensitivity of PI, the catechin biosynthesis network in fresh tea leaves could be clearly illustrated, indicating the potential applicability of imprint DESI/PI in exploring the sites and pathways of plant metabolic conversion.

Effects of dopants in the imaging of mouse brain by desorption electrospray ionization/post-photoionization mass spectrometry.

Desorption electrospray ionization/post-photoionization (DESI/PI) is a newly developed ionization method by the combination of DESI and post-photoionization for the simultaneous imaging of polar and nonpolar compounds in biological tissues. Dopants are of great importance in DESI/PI for the enhancement of signal intensities through ion-molecule reactions. In this work, to evaluate the performance of dopants in DESI/PI, an efficient homogenate model was developed, and four kinds of dopants (toluene, chlorobenzene, bromobenzene, and anisole) were tested using homogenate of mouse brain tissue as target sample. The influences of the dopants on the signal enhancements of different compounds were explained reasonably by the ionization mechanism. Then, the dopants with their optimum volume contents were applied to the mass spectrometry imaging (MSI). For a comprehensive imaging of various compounds with different polarities, methanol/toluene/formic acid (7:3:0.1) was chosen as the best choice. Finally, the stronger quantitative ability of DESI/PI with toluene as dopant for a few compounds in mouse brain tissue was demonstrated.

Recent Advances of Ambient Mass Spectrometry Imaging and Its Applications in Lipid and Metabolite Analysis.

Ambient mass spectrometry imaging (AMSI) has attracted much attention in recent years. As a kind of unlabeled molecular imaging technique, AMSI can enable in situ visualization of a large number of compounds in biological tissue sections in ambient conditions. In this review, the developments of various AMSI techniques are discussed according to one-step and two-step ionization strategies. In addition, recent applications of AMSI for lipid and metabolite analysis (from 2016 to 2021) in disease diagnosis, animal model research, plant science, drug metabolism and toxicology research, etc., are summarized. Finally, further perspectives of AMSI in spatial resolution, sensitivity, quantitative ability, convenience and software development are proposed.

Online Monitoring the Key Intermediates and Volatile Compounds Evolved from Green Tea Roasting by Synchrotron Radiation Photoionization Mass Spectrometry.

Online monitoring of the volatile compounds during the tea roasting process is crucial to find the optimum roasting conditions and improve the quality of green tea. In this work, synchrotron radiation photoionization mass spectrometry (SR-PIMS) was utilized to online monitor the evolved gaseous compounds during the tea roasting process. By virtue of "soft" ionization and fast data acquisition characteristics of SR-PIMS, dozens of aroma compounds including alcohols, aldehydes, furans, and nitrogen- and sulfur-containing species were detected and identified in real time. Moreover, 5-hydroxymethylfurfural (5-HMF), the key intermediate of Maillard reactions, was found with high sensitivity. Evolution processes of all the products could be observed via the time- and temperature-resolved profiles in N2 and the air. Dehydration was found to be the first step during roasting. Oxygen in the air was found to accelerate the formation rate of various stable species and intermediates in the course of the thermal treatment of fresh green tea. The formation mechanisms of evolved compounds such as three sulfur-containing compounds, i.e., dimethyl sulfide, hydrogen sulfide, and methanethiol, could be proposed according to the step-by-step formation process. The time-resolved results were demonstrated to be applicable in the evaluation of different roasting processes by statistical analysis. The optimum tea roasting temperature and duration are proposed to be around 200 °C and 1000 s.

Cholesterol was identified as a biomarker in human melanocytic nevi using DESI and DESI/PI mass spectrometry imaging.

The rapid differentiation between diseased tissue and healthy normal tissue is of great importance for the intraoperative diagnosis. Herein, desorption electrospray ionization (DESI) and DESI/post-photoionization (DESI/PI) mass spectrometry imaging were combined to in situ visualize the distribution of biochemicals within the tissue regions of human melanocytic nevi under the ambient condition with a spatial resolution of around 200 µm. Plenty of polar and nonpolar lipids were found to be specifically distributed in melanocytic nevi with statistical significance and could be used to differentiate the healthy normal tissue and melanocytic nevi. Cholesterol was further confirmed to be a potential biomarker for melanocytic nevi diagnosis by multivariate statistical analysis and immunohistochemistry of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and translocator protein (TSPO) enzymes. This work provides a visual way for the diagnosis of human melanocytic nevi by lipid profiling, which benefits the understanding of the pathological mechanism of melanocytic nevi and provides a new insight to control melanin growth from the synthesis, transport, and metabolism of cholesterol.