Top–down glycolipidomics: fragmentation analysis of ganglioside oligosaccharide core and ceramide moiety by chip‐nanoelectrospray collision‐induced dissociation MS2–MS6

We developed a straightforward approach for high‐throughput top–down glycolipidomics based on fully automated chip‐nanoelectrospray (nanoESI) high‐capacity ion trap (HCT) multistage mass spectrometry (MSⁿ) by collision‐induced dissociation (CID) in the negative ion mode. The method was optimized and tested on a polysialylated ganglioside fraction (GT1b), which was profiled by MS¹ and sequenced in tandem MS up to MS⁶ in the same experiment. Screening of the fraction in the MS¹ mode indicated the occurrence of six [M ? 2H]^2? ions which, according to calculation, support 13 GT1 variants differing in their relative molecular mass due to dissimilar ceramide (Cer) constitutions. By stepwise CID MS²–MS⁵ on the doubly charged ion at m/z 1077.20 corresponding to a ubiquitous GT1b structure, the complete characterization of its oligosaccharide core including the identification of sialylation sites was achieved. Structure of the lipid moiety was further elucidated by CID MS⁶ analysis carried out using the Y₀ fragment ion, detected in MS⁵, as a precursor. MS⁶ fragmentation resulted in a pattern supporting a single ceramide form having the less common (d20 : 1/18 : 0) configuration. The entire top–down experiment was performed in a high‐throughput regime in less than 3 min of measurement, with an analysis sensitivity situated in the subpicomolar range. Copyright © 2009 John Wiley & Sons, Ltd.     

Determination of ganglioside composition and structure in human brain hemangioma by chip-based nanoelectrospray ionization tandem mass spectrometry

We report here on a preliminary investigation of ganglioside composition and structure in human hemangioma, a benign tumor in the frontal cortex (HFC) in comparison to normal frontal cortex (NFC) tissue using for the first time advanced mass spectrometric methods based on fully automated chip-nanoelectrospray (nanoESI) high-capacity ion trap (HCT) and collision-induced dissociation (CID). The high ionization efficiency, sensitivity and reproducibility provided by the chip-nanoESI approach allowed for a reliable MS-based ganglioside comparative assay. Unlike NFC, ganglioside mixture extracted from HFC was found dominated by species of short glycan chains exhibiting lower overall sialic acid content. In HFC, only GT1 (d18:1/20:0), and GT3 (d18:1/25:1) polysialylated species were detected. Interestingly, none of these trisialylated forms was detected in NFC, suggesting that such components might selectively be associated with HFC. Unlike the case of previously investigated high malignancy gliosarcoma, in HFC one modified O-Ac-GD2 and one modified O-Ac-GM4 gangliosides were observed. This aspect suggests that these O-acetylated structures could be associated with cerebral tumors having reduced malignancy grade. Fragmentation analysis by CID in MS² mode using as precursors the ions corresponding to GT1 (d18:1/20:0) and GD1 (d18:1/20:0) provided data corroborating for the first time the presence of the common GT1a and GT1b isomers and the incidence of unusual GT1c and GT1d glycoforms in brain hemangioma tumor.      

Chip electrospray mass spectrometry for carbohydrate analysis

Currently two types of chip systems are used in conjunction with MS: out-of-plane devices, where hundreds of nozzles, nanospray emitters are integrated onto a single silicon substrate from which electrospray is established perpendicular to the substrate, and planar microchips, embedding a microchannel at the end of which electrospray is generated in-plane, on the edge of the microchip. In the last two years, carbohydrate research greatly benefited from the introduction and implementation of the chip-based MS. In two laboratories the advantages of the chip electrospray in terms of ionization efficiency, sensitivity, reproducibility, quality of data in combination with high mass accuracy, and resolution of detection were systematically explored for several carbohydrate classes: O- and N-glycopeptides, oligosaccharides, gangliosides and glycoprotein-derived O- and N-glycans, and glycopeptides. The current state-of-the-art in interfacing the chip electrospray devices to high-performance MS for carbohydrate analysis, and the particular requirements for method optimization in both positive and negative ion modes are reviewed here. The recent applications of these miniaturized devices and their general potential for glycomic-based surveys are highlighted.     

Profiling and sequencing of gangliosides from human caudate nucleus by chip‐nanoelectrospray mass spectrometry

Gangliosides (GGs), sialic acid‐containing glycosphingolipids are involved in many brain functions at the cell and molecular level. Compositional and structural elucidation of GGs in mixtures extracted from human brain is essential for correlating their profile with the specialized function of each brain area in health and disease. As a part of our ongoing study on GG expression and structure in different healthy and diseased brain regions, in this work, a preliminary investigation of GGs in a specimen of human caudate nucleus (CN) was carried out using an advanced mass spectrometry (MS) technique. By chip‐nanoelectrospray MS performed on a NanoMate robot coupled to a high capacity ion trap instrument, 81 GG components were detected in human CN in only 1.5 min of signal acquisition. Although the native GG mixture from CN was found dominated by mono‐, di‐ and trisialylated GGs with a slight dominance of disialylated forms (GD), four tetrasialylated structures (GQ) and two pentasialylated (GP) species were also identified. Additionally, species with unusually long fatty acid chains, exceeding 30 carbon atoms in their ceramide (Cer) composition, and several glycoforms modified by fucosyl (Fuc), O‐acetyl (O‐Ac) and/or lactonization were discovered. By tandem MS (MS²) using collision‐induced dissociation, two atypical mono and disialylated species with long‐chain fatty acids in their Cer could be confirmed and structurally characterized. These results may be a starting point for new GG‐based approaches in the study of CN functions and ethiopathogenesis of CN‐related neurodegenerative disorders. Copyright © 2012 John Wiley & Sons, Ltd.     

Quantum phase transitions in mesoscopic systems

Quantum phase transitions in mesoscopic systems are studied. It is shown that the main features of phase transitions, defined for infinite number of particles, N--> infinity, persist even for moderate N approximately 10. A Landau analysis of first order transitions is done and a "critical" exponent at the spinodal point is defined. Two order parameters are introduced to distinguish first from second order transitions. Applications to atomic nuclei, molecules, atomic clusters, and finite polymers are mentioned. Experimental evidence in atomic nuclei is presented.     

Coulomb excitation of radioactive 132,134,136Te beams and the low B(E2) of 136Te

The B(E2;0(+)-->2+) values for the first 2+ excited states of neutron-rich 132,134,136Te have been measured using Coulomb excitation of radioactive ion beams. The B(E2) values obtained for 132,134Te are in excellent agreement with expectations based on the systematics of heavy stable Te isotopes, while that for 136Te is unexpectedly small. These results are discussed in terms of proton-neutron configuration mixing and shell-model calculations using realistic effective interactions.