Abstract
Small-molecule (e.g., metabolite) and low-abundance neuropeptide analyses by mass spectrometry (MS) represent important research directions and have witnessed tremendous growth and developments during past decades. With innate advantages of MS and gentle nature of soft ionization techniques including electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI), profiling and visualization of these bioactive metabolites and neuropeptides have undergone technological advancements that can be applied to real biological systems, although numerous challenges still exist. We herein present a rapid and efficient strategy to improve both metabolite and neuropeptide analysis, the nanosecond photochemical reaction (nsPCR)-enabled fast chemical derivatization. Amine-directed chemoselectivity facilitates the rapid tagging on amine-containing metabolites and neuropeptides, resulting in improved detection sensitivity. Additionally, the nsPCR generates a localized pH jump zone and enables localized thermophoresis at nanosecond timescale which benefits on-demand matrix removal during MALDI-MS identification and visualization of low-abundance biomolecules. A step-by-step nsPCR experimental protocol is introduced in detail herein for both spot analysis and imaging analysis, followed by suggestions for data analysis to ensure successful application of the nsPCR strategy.
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Acknowledgments
This work was funded in part by NIH (R56MH110215, R01DK071801, and P01CA250972) and NSF (CHE-1710140). The MALDI Orbitrap instrument and Q Exactive Orbitrap instrument were purchased through the support of an NIH shared instrument grant S10RR029531. LL acknowledges a Vilas Distinguished Achievement Professorship and the Charles Melbourne Johnson Distinguished Chair Professorship with funding provided by the Wisconsin Alumni Research Foundation and University of Wisconsin-Madison School of Pharmacy. GL thanks the funding support by the Fundamental Research Funds for the Central Universities (Nankai University, 020/63213057) and by the National Natural Science Foundation of China (22104064). The authors would like to thank Drs. Eugene Moskovets and Vladimir Doroshenko from MassTech Inc. for access to a SubAP MALDI source and technical support.
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Li, G., Liu, Y., Li, L. (2022). Nanosecond Photochemical Reaction (nsPCR) for Enhanced Mass Spectrometric Identification, Quantification, and Visualization of Metabolites and Neuropeptides. In: Lee, YJ. (eds) Mass Spectrometry Imaging of Small Molecules. Methods in Molecular Biology, vol 2437. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2030-4_10
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DOI: https://doi.org/10.1007/978-1-0716-2030-4_10
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