Lipidomics of intact mitochondria by MALDI-TOF/MS

Abstract

onstrated for the analysis of lipid extracts from different biological materials ( 14-18 ), but the most promising advantage of the MALDI-TOF/MS technique turned out to be the possibility to perform lipid analysis avoiding extraction and/or separation steps.Recent advances in MALDI-TOF/MS techniques of lipid analysis have led to the direct coupling of , as well as to the direct analysis of tissue slices with the MALDI-MSI ( 23-25 ). In both cases, a plate containing the sample is coated with the matrix solution (silica plate or tissue on microscope glass plate, respectively), attached onto the MALDI target, and directly scanned with the laser to obtain a fast MS analysis of the lipids on the plate. Both these methods allow a fast and convenient lipid analysis, skipping extraction steps that are usually required for these kinds of analyses.In 2010 we developed a protocol that allows the direct lipid analysis by MALDI mass spectrometry in intact archaeal membranes without prior extraction/separation steps: lyophilized archaeal membranes were ground with 9-aminoacridine (9-AA) as dry matrix, and the powder mixture was crushed in a mechanical die press to form a thin pellet; small pieces of the pellet were then attached onto the MALDI target, and the archaeal membrane lipid profi le was directly achieved ( 26 ).Another new protocol for the direct lipid fi ngerprinting with MALDI-TOF/MS was published by Ferreira et al. in 2010 ( 27 ) Historically lipid profi ling of biological samples has been achieved with preliminary isolation of tissues, cells, membranes or organelles, followed by lipid extraction and purifi cation with chromatographic techniques, such as HPLC or (HP)TLC ( 1, 2 ), and by lipid characterization and/or quantitation with gas chromatography (GC) ( 3 ), GC-mass spectrometry (MS) ( 4 ), fast atom bombardment (FAB)-MS ( 5-7 ), , and/or NMR analyses ( 13 ).In the last decade, the capabilities of MALDI time-offl ight (TOF)/MS in lipid analysis have been largely dem-

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2011–2012

“…The results compare favorably with those from previous experiments involving TLC separations and has also been applied to intact membranes from the bacterium Paracoccus denitrificans (Angelini et al, 2012b).…”

Cardiolipin fingerprinting of leukocytes by MALDI-TOF/MS as a screening tool for Barth syndrome

“…Importantly, we found that the phosphatidylcholine-tolysophosphatidylcholine ratio could be easily determined by MALDI-TOF/MS analyses of native blood serum and followed as a clinical parameter in various diseases ( 32 ). Relevantly, we also reported that CL could be easily detected in direct MALDI-TOF/MS total lipid profi les, by using 9-aminoacridine (9-AA) as matrix ( 31 ), even in the lipid fi ngerprint of whole cells.…”

“…Indeed MALDI-TOF/ MS allows the direct analysis of native samples, skipping timeconsuming steps of standard lipid extraction and separation procedures. The success of our experimental approach is due to the use of 9-AA as matrix, which is particularly suitable for CLs detection ( 31 ), even when these species are only minor components in complex lipid profi les.…”

“…It has been shown that this emerging analytical technique can be used to directly acquire lipid profi les of biological samples, thus skipping extraction and separation steps (30)(31)(32)(33). We successfully adopted this experimental approach to describe the lipid composition of bacterial and mitochondrial membranes ( 30,31,33 ) and to detect lipid markers in blood serum ( 32 ). Importantly, we found that the phosphatidylcholine-tolysophosphatidylcholine ratio could be easily determined by MALDI-TOF/MS analyses of native blood serum and followed as a clinical parameter in various diseases ( 32 ).…”

Comprehensive approach to the quantitative analysis of mitochondrial phospholipids by HPLC–MS

“…Therefore, analysis of CL, PG, and MLCL will provide mechanistic information concerning the biosynthesis of CL. Recently, MALDI-TOF analysis of phospholipids in intact mitochondria was reported [12]. The method provides a quick and simple procedure and detection of various lipids classes, including these low abundance phospholipids.…”