Abstract
The application of desorption electrospray ionization mass spectrometry (DESI-MS) and dried blood spot (DBS) sampling has been successfully implemented several times. However, the difficulty of combining DBS sampling with DESI-MS is still the carrier material used for the blood samples. In this study, a new, easily obtained, and cost-effective carrier substrate for dried plasma spot (DPS) sampling and DESI-MS analysis and its application in phospholipidomics studies was described. First, the effects of several carrier materials, including cellulose-based materials (31 ET paper and filter paper) and non-cellulose-based materials (PARAFILM and its shape-modified material, PTFE-printed glass slide and polyvinylidene fluoride film), were tested. Second, a method combining DPS sampling with DESI-MS for phospholipidomics analysis was established, and parameters affecting compound signal intensities, such as sample volume and sprayer solvent system, were optimized. In conclusion, the total signal intensity obtained from shape-modified PARAFILM was the strongest. The suitable plasma sample volume deposited on PARAFILM carriers was 5 μl, and acetonitrile (ACN) was recommended as the optimal spray solvent for phospholipid (PL) profiling. Repeatability (87.5% of compounds with CV < 30%) and stability for data acquisition (48 h) were confirmed. Finally, the developed method was applied in phospholipidomics analysis of schistosomiasis, and a distinguished classification between control mice and infected mice was observed by using multivariate pattern recognition analysis, confirming the practical application of this new carrier material for DPS sampling and DESI-MS analysis. Compared with a previously reported method, the rapid metabolomics screening approach based on the implementation of DPS sampling coupled with the DESI-MS instrument developed in this study has increased analyte sensitivity, which may promote its further application in clinical studies.
Highlights
Phospholipids (PLs), as the major components of the cell membrane, exist in both eukaryotic and prokaryotic organisms (Hanahan, 1997); PLs play important roles in controlling and regulating cellular function and participate in many cellular biological processes (Subramaniam et al, 2011)
As the mass spectra acquired from the strongest scan point on total ion chromatography of each material showed in Figure 1A, we observed that the whole signal intensities of compounds in non-cellulose-based carrier materials were commonly higher than those in cellulosed-based carrier materials
We modified the shape of PARAFILM due to its extensibility; that is, we imprinted a certain number of circular grooves with 0.1 mm in depth and 4 mm in diameter in PARAFILM, on which dried plasma spot (DPS) could be fixed firmly in the course of desorption electrospray ionization mass spectrometry (DESI-mass spectrometry (MS)) analysis
Summary
Phospholipids (PLs), as the major components of the cell membrane, exist in both eukaryotic and prokaryotic organisms (Hanahan, 1997); PLs play important roles in controlling and regulating cellular function and participate in many cellular biological processes (Subramaniam et al, 2011). Over the past few decades, PLs have generally been obtained through liquid-liquid extraction methods (Bligh and Dyer, 1959) or protein precipitation methods (Sarafian et al, 2014) to detect the metabolic fluctuation in PLs in vivo. These methods both include deproteinization, solvent extraction, centrifugation, and other preprocessing steps, followed by time-consuming chromatographic separation. These procedures limit sample throughput and inevitably lead to the loss of metabolites. It is necessary to develop a high-throughput phospholipidomics method to profile the changes in PLs in organisms
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