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. 2011 Jan 15;408(2):289-96.
doi: 10.1016/j.ab.2010.09.017. Epub 2010 Sep 22.

Analysis of lipids from crude lung tissue extracts by desorption electrospray ionization mass spectrometry and pattern recognition

Franco Basile  1 , Tamara Sibray, John T Belisle, Richard A Bowen
Affiliations

Analysis of lipids from crude lung tissue extracts by desorption electrospray ionization mass spectrometry and pattern recognition

Franco Basile et al. Anal Biochem. .

Abstract

A method is described using desorption electrospray ionization (DESI) mass spectrometry (MS) to obtain phospholipid mass spectral profiles from crude lung tissue extracts. The measured DESI mass spectral lipid fingerprints were then analyzed by unsupervised learning principal components analysis (PCA). This combined approach was used to differentiate the effect(s) of two vaccination routes on lipid composition in mouse lungs. Specifically, the two vaccination routes compared were intranasal (i.n.) and intradermal (i.d.) inoculation of the Francisella tularensis live vaccine strain (Ft-LVS). Lung samples of control and LVS-inoculated mice were quickly extracted with a methanol/chloroform solution, and the crude extract was directly analyzed by DESI-MS, with a total turnaround time of less than 10 min/sample. All of the measured DESI mass spectra (in both positive and negative ion mode) were compared via PCA, resulting in clear differentiation of mass spectral profiles of i.n.-inoculated mouse lung tissues from those of i.d.-inoculated and control mouse lung tissues. Lipid biomarkers responsible for sample differentiation were identified via tandem MS (MS/MS) measurements or by comparison with mass spectra of lipid standards. The DESI-MS approach described here provided a practical and rapid means to analyze tissue samples without extensive extractions and solvent changes.

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Figures

Figure. 1
Figure. 1
Mouse lung sample preparation procedure for DESI-MS and time comparison with infusion ESI-MS analysis.
Figure. 2
Figure. 2
Direct analysis of mouse lung tissue lipid extract (i.n. LVS inoculated) with (a) positive ion DESI-MS and (b) direct infusion positive ion ESI-MS. For the ESI-MS analysis, the sample solvent was changed from chloroform:methanol to 50% methanol water (1 % acetic acid) in order to obtain comparable results as in the DESI-MS analysis.
Figure. 3
Figure. 3
Negative ion mode DESI-mass spectra of crude extract of mouse lung tissue (a) i.d. LVS (b) i.n. LVS inoculated mouse lungs (tentative assignments of sn1 and sn2 groups were based on MS/MS analyses).
Figure. 4
Figure. 4
Positive ion DESI mass spectra of crude extracts from (a) i.d. LVS and (b) i.n. LVS inoculated mouse lungs (tentative assignments of GPCho sn-1 and sn-2 groups were based on MS/MS analyses).
Figure. 5
Figure. 5
Principal component analysis of negative ion DESI-mass spectra of mice crude lung tissue (a) Score plot and (b) Loading plot
Figure. 6
Figure. 6
Principal component analysis of positive ion DESI-mass spectra of mice crude lung tissue (a) Scores plot (b) Loading plot.
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