Quantitative Imprint Mass Spectrometry Imaging of Endogenous Ceramides in Rat Brain Tissue with Kinetic Calibration
- 7 April 2020
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 92 (9), 6613-6621
- https://doi.org/10.1021/acs.analchem.0c00392
Abstract
Quantitative mass spectrometry imaging (MSI) is an effective technique for determining the spatial distribution of molecules in a variety of sample types; however, the quality of the ion signals is related to the chemical and morphological properties of the tissue and the targeted analyte(s). Issues may arise with the incorporation of standards into the tissue at repeatable, welldefined concentrations, as well as with the extraction and incorporation of endogenous analytes versus standards from tissue into the matrix To address these concerns, we combine imprint MSI (iMSI) with kinetic calibration and use it to quantify lipids in rat brain tissue samples. Briefly, tissues were imprinted on slides coated with a dopamine-modified TiO, monolith pretreated with analyte standards, resulting in the adsorption of endogenous analytes onto the coating and desorption of standards into the tissue. The incorporation of standards into the tissue enabled quantification of the measured analytes using kinetic calibration. Moreover, matrix effects were reduced, and the intensities of analyte standard signals became more uniform. The symmetry of the adsorption of endogenous ceramides and the desorption of ceramide standards suggest that the content of adsorbed endogenous ceramide can be determined by measuring the content of desorbed ceramide standard. Using kinetic calibration, endogenous ceramide concentrations were calculated for a range of dry and wet tissue imprinting conditions and compared to quantitative MSI using a standard spiking approach. We validated the relative quantitative values from iMSI using liquid chromatography tandem mass spectrometry (LC-MS/MS) and found that the ratios from iMSI as compared to LC-MS/MS were in the range of 70-200% over the concentration range of endogenous ceramides; the correlation coefficient between iMSI and LC-MS/MS was over 0.9 (Pearson's r), while the relative recoveries via traditional standard spiking were between 200% and 5000% depending on the brain region and sample preparation conditions.Funding Information
- National Institute on Drug Abuse (P30DA018310)
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