Direct analysis of glycolipids on thin-layer plates by matrix-assisted secondary ion mass spectrometry: Application for glycolipid storage disorders

Kushi, Yasunori; Rokukawa, Chiaki; Handa, Shizuo

doi:10.1016/0003-2697(88)90375-2

Cited by 40 publications

(17 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The mass spectra obtained were comparable to those acquired by conventional methods (Kushi et al, 1983). Applications of TLC-SI-MS for GSL analysis of lipid extracts from (1) organs of patients suffering from GSL storage diseases such as Gaucher's, Tay-Sachs, and Fabry's disease and (2) human erythrocytes of blood group A-type have been later reported by Kushi, Handa, and co-workers (Kushi, Rokukawa, & Handa, 1988;Kushi et al, 1990).…”

Section: Early Tlc-ms Experimentsmentioning

confidence: 87%

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Müthing

Distler

2009

Mass Spectrom. Rev.

View full text Add to dashboard Cite

Glycosphingolipids (GSLs), composed of a hydrophilic carbohydrate chain and a lipophilic ceramide anchor, play pivotal roles in countless biological processes, including infectious diseases and the development of cancer. Knowledge of the number and sequence of monosaccharides and their anomeric configuration and linkage type, which make up the principal items of the glyco code of biologically active carbohydrate chains, is essential for exploring the function of GSLs. As part of the investigation of the vertebrate glycome, GSL analysis is undergoing rapid expansion owing to the application of novel biochemical and biophysical technologies. Mass spectrometry (MS) takes part in the network of collaborations to further unravel structural and functional aspects within the fascinating world of GSLs with the ultimate aim to better define their role in human health and disease. However, a single-method analytical MS technique without supporting tools is limited yielding only partial structural information. Because of its superior resolving power, robustness, and easy handling, high-performance thin-layer chromatography (TLC) is widely used as an invaluable tool in GSL analysis. The intention of this review is to give an insight into current advances obtained by coupling supplementary techniques such as TLC and mass spectrometry. A retrospective view of the development of this concept and the recent improvements by merging (1) TLC separation of GSLs, (2) their detection with oligosaccharide-specific proteins, and (3) in situ MS analysis of protein-detected GSLs directly on the TLC plate, are provided. The procedure works on a nanogram scale and was successfully applied to the identification of cancer-associated GSLs in several types of human tumors. The combination of these two supplementary techniques opens new doors by delivering specific structural information of trace quantities of GSLs with only limited investment in sample preparation.

show abstract

Section: Early Tlc-ms Experimentsmentioning

confidence: 87%

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Müthing

Distler

2009

Mass Spectrom. Rev.

View full text Add to dashboard Cite

show abstract

“…Another improvement in structural determination of GSLs was obtained using high performance FAB-MS/MS. [12][13][14][15] The introduction of electrospray ionization (ESI) has opened another promising area for mass spectrometric analyses of complex carbohydrates. The application of the ESI technique in the positive ion mode to analyse neutral native glycosphingolipids was proposed by Siuzdak et al, 16 who showed that ESI results in the production of singly and doubly charged molecular ions with monovalent cations such as Na as adducts.…”

Section: Received 22 August 1997; Revised 18 March 1998; Accepted 20 mentioning

confidence: 99%

“…Fast atom bombardment (FAB) ionization of native GSLs shows mainly intense molecular and sequence fragment ions, thus simplifying the structure elucidation of pure compounds. [9][10][11][12] For analysis of mixtures, interpretation of spectra containing molecular and fragment ions from several GSLs with different carbohydrate chain lengths is a problem. Another improvement in structural determination of GSLs was obtained using high performance FAB-MS/MS.…”

Section: Received 22 August 1997; Revised 18 March 1998; Accepted 20 mentioning

confidence: 99%

Electrospray ionization and collision-induced dissociation time-of-flight mass spectrometry of neutral glycosphingolipids

Olling

Breimer

Peltomaa

et al. 1998

Rapid Commun. Mass Spectrom.

View full text Add to dashboard Cite

A series of native naturally occurring neutral glycosphingolipids has been analysed by electrospray ionization tandem mass spectrometry using a hybrid magnetic sector-TOF instrument. The collision-induced dissociation products of precursor ions were detected by an orthogonal acceleration time-of-flight mass spectrometer as the second analyser. Glycosphingolipids, with mono- to hexa-saccharide chain lengths with different ceramide constituents, were studied. The result of electrospray ionization in the positive ion mode generally showed singly charged molecular ions with Na+ as adduct, [M + Na]+. The sensitivity of the electrospray ionization was greatly enhanced by addition of NaCl, LiCl (forming [M + Li]+) or KCl (yielding [M + K]+) to the sample. A comparison between the collision-induced dissociation of precursor molecular ions of monoglycosylceramides, using Na+, Li+ and K+ as adducting species, showed that the intensity of the fragment ions and the extent of the daughter ion fragmentation of the molecular ions, are dependent on the type of adduct used. The daughter ion spectra of Li+ adduct ions showed intense sequence fragment ions, both of the saccharide chain and the ceramide moiety, and were superior to those obtained using Na+ or K+. The collision-induced dissociation spectra of the [M + Li]+ ions, of glycosphingolipids containing di- to hexasaccharides, are also presented. Proposed possible fragments, resulting from the CID of the molecular ions [M + Li]+ of monoglycosylceramides, are shown.

show abstract

“…Mass spectrometry (MS) is routinely used to define the primary structure of purified GSLs or GSLs in mixtures 11. Improved techniques were the direct desorption of separated GSLs from thin‐layer chromatograms by fast‐atom bombardment mass spectrometry (FAB‐MS),12 matrix‐assisted secondary ion mass spectrometry (SI‐MS),13 and direct matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) of GSLs from HPTLC plates 14, 15…”

mentioning

confidence: 99%

Application of combined high‐performance thin‐layer chromatography immunostaining and nanoelectrospray ionization quadrupole time‐of‐flight tandem mass spectrometry to the structural characterization of high‐ and low‐affinity binding ligands of Shiga toxin 1

Meisen

Friedrich

Karch

et al. 2005

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

Shiga toxin 1 (Stx1) represents an AB5 toxin produced by enterohemorrhagic Escherichia coli, which cause gastrointestinal diseases in humans that are often followed by potentially fatal systemic complications, such as acute encephalopathy and hemolytic uremic syndrome. The expression of the preferential Stx1 receptor, Gb3Cer/CD77 (Gal alpha1-4Gal beta1-4Glc beta1-1Cer), is one of the primary determinants of susceptibility to tissue injury. Due to the clinical importance of this life-threatening toxin, a combined strategy of preparative high-performance thin-layer chromatography (HPTLC) overlay assay and mass spectrometry was developed for the detection and structural characterization of Stx1-binding glycosphingolipids (GSLs). A preparation of neutral GSLs from human erythrocytes, comprising 21.4% and 59.1% of the high- and low-affinity Stx1-binding ligands Gb3Cer/CD77 and Gb4Cer, respectively, was separated on silica gel precoated HPTLC plates and probed for the presence of Stx1 receptors. Stx1 positive on the one hand and anti-Gb3Cer/CD77 and anti-Gb4Cer antibody positive bands from parallel reference runs on the other hand were extracted with chloroform/methanol/water (30/60/8, v/v/v). These crude extracts were used without any further purification for a detailed structural analysis by nanoelectrospray ionization quadrupole time-of-flight mass spectrometry (nanoESI-QTOF-MS) in the negative ion mode. In all extracts investigated, neutral GSLs were detected as singly charged deprotonated molecular ions, [M-H]-, and neither buffer-derived salt adducts nor coextracted contaminants from the overlay assay procedure or the silica gel layer were observed. For the structural characterization of Stx1- and antibody-binding GSLs low-energy collision-induced dissociation (CID) was applied to high and low abundant receptor species of the crude extracts. All MS/MS spectra obtained contained full series of Y-type ions, B-type ions and additional ions generated by ring cleavages of the sugar moiety. Only analytical quantities in the microgram scale of a single GSL species within the complex GSL mixture were required for the structural MS characterization of Stx1 ligands as Gb3Cer/CD77 and Gb4Cer. This effective combined HPTLC/MS procedure offers a broad range of applications, not only for toxins of bacterial origin, but also for any GSL-binding agents such as plant-derived lectins or human proteins with yet unknown binding specificities.

show abstract

Direct analysis of glycolipids on thin-layer plates by matrix-assisted secondary ion mass spectrometry: Application for glycolipid storage disorders

Cited by 40 publications

References 22 publications

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Electrospray ionization and collision-induced dissociation time-of-flight mass spectrometry of neutral glycosphingolipids

Application of combined high‐performance thin‐layer chromatography immunostaining and nanoelectrospray ionization quadrupole time‐of‐flight tandem mass spectrometry to the structural characterization of high‐ and low‐affinity binding ligands of Shiga toxin 1

Contact Info

Product

Resources

About