Analysis of Gangliosides Directly from Thin-Layer Chromatography Plates by Infrared Matrix-Assisted Laser Desorption/Ionization Orthogonal Time-of-Flight Mass Spectrometry with a Glycerol Matrix

Dreisewerd, Klaus; Müthing, Johannes; Rohlfing, Andreas; Meisen, Iris; Vukelić, Željka; Peter‐Katalinić, Jasna; Hillenkamp, Franz; Berkenkamp, Stefan

doi:10.1021/ac048373w

Cited by 119 publications

(117 citation statements)

References 41 publications

(47 reference statements)

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“…Furthermore, continuous improvements in the methodology described here and the alternative use of an infrared laser for desorption can be expected to lower the detection limits. Experiments of the latter type are underway elsewhere [39]. …”

Section: Resultsmentioning

confidence: 99%

Ganglioside analysis by thin-layer chromatography matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry

Ivleva

Sapp

O’Connor

et al. 2005

J. Am. Soc. Mass Spectrom.

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Metastable decomposition of ions generated in matrix-assisted laser desorption/ionization (MALDI) mass spectrometers complicates analysis of biological samples that have labile bonds. Recently, several academic laboratories and manufacturers of commercial instruments have designed instruments that introduce a cooling gas into the ion source during the MALDI event and have shown that the resulting vibrational cooling stabilizes these labile bonds. In this study, we compared stabilization and detection of desorbed gangliosides on a commercial orthogonal time-of-flight (oTOF) instrument with results we reported previously that had been obtained on a home-built Fourier transform mass spectrometer. Decoupling of the desorption/ ionization from the detection steps resulted in an opportunity for desorbing thin-layer chromatography (TLC)-separated gangliosides directly from a TLC plate without compromising mass spectral accuracy and resolution of the ganglioside analysis, thus coupling TLC and oTOF mass spectrometry. The application of a declustering potential allowed control of the matrix cluster and matrix adduct formation, and, thus, enhanced the detection of the gangliosides. during matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analyses of molecules having labile bonds. Although this phenomenon can be used for obtaining analyte structural information for small peptides, oligosaccharides, and some glycolipids [1-5] for larger or more fragile molecules, it can limit the mass accuracy and resolution and complicate interpretation of the spectra of heterogeneous samples. Thus, it is important to develop methods that allow control of the extent of metastable decay. In addition to the pressure of the background gas and extraction field strength in the MALDI technique, the major factors affecting metastable fragmentation are laser wavelength/pulse width and the choice of the MALDI matrix [6,7]. In addition, the presence of electrons may play a role in analyte fragmentation and may cause suppression of multiply charged ions in a spectrum [8,9].Control of the metastable fragmentation in MALDI by increasing the pressure in the ionization source and reduction of the ions internal energy was first introduced by Loboda et al. on orthogonal time-of-flight (oTOF) instruments [10] and later extended to Fourier transform mass spectrometry (FTMS) [11] and to the operation at atmospheric pressure on various types of instruments [12]. These methods have since been used for stabilization of glycolipids [13,14] and peptides and proteins [11,15]. The term vibrational cooling (VC) MALDI recently has been introduced to describe desorption in the pressure range where cooling of the excess bond energy is achieved [12]. Stabilization of the labile bonds under these conditions allows for control of the extent of fragmentation, which is particularly

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Section: Resultsmentioning

confidence: 99%

Ganglioside analysis by thin-layer chromatography matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry

Ivleva

Sapp

O’Connor

et al. 2005

J. Am. Soc. Mass Spectrom.

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“…Furthermore, recent applications of AP-MALDI using tightly focused laser beams (spot size ~5-7 µm) has allowed the lipid composition of single cells, including HeLa cells [101] when only a few unique lipids were present at a given sampling spot. Glycerol adducts have also been observed for GL analysis in negative ion mode [114], and are attributed to the use of an elevated pressure ionization source and associated collisional cooling of the adducts.…”

Section: Atmospheric Pressure Maldi (Ap-maldi)mentioning

confidence: 99%

“…In addition, the greater laser penetration depth means IR-MALDI is also well suited for analysis of TLC plates using a glycerol matrix, while there are also numerous reports of successful application of intermediate and atmospheric pressure sources [107,109,110,[113][114][115].…”

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confidence: 99%

Surface analysis of lipids by mass spectrometry: More than just imaging

Ellis

Brown

Panhuis

et al. 2013

Progress in Lipid Research

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Mass spectrometry is now an indispensable tool for lipid analysis and is arguably the driving force in the renaissance of lipid research. In its various forms, mass spectrometry is uniquely capable of resolving the extensive compositional and structural diversity of lipids in biological systems. Furthermore, it provides the ability to accurately quantify molecular-level changes in lipid populations associated with changes in metabolism and environment; bringing lipid science to the "omics" age. The recent explosion of mass spectrometry-based surface analysis techniques is fuelling further expansion of the lipidomics field. This is evidenced by the numerous papers published on the subject of mass spectrometric imaging of lipids in recent years. While imaging mass spectrometry provides new and exciting possibilities, it is but one of the many opportunities direct surface analysis offers the lipid researcher. In this review we describe the current state-ofthe-art in the direct surface analysis of lipids with a focus on tissue sections, intact cells and thin-layer chromatography substrates. The suitability of these different approaches towards analysis of the major lipid classes along with their current and potential applications in the field of lipid analysis are evaluated. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 AbstractMass spectrometry is now an indispensable tool for lipid analysis and is arguably the driving force in the renaissance of lipid research. In its various forms, mass spectrometry is uniquely capable of resolving the extensive compositional and structural diversity of lipids in biological systems. Furthermore, it provides the ability to accurately quantify molecular-level changes in lipid populations associated with changes in metabolism and environment;bringing lipid science to the "omics" age. The recent explosion of mass spectrometry-based surface analysis techniques is fuelling further expansion of the lipidomics field. This is evidenced by the numerous papers published on the subject of mass spectrometric imaging of lipids in recent years. While imaging mass spectrometry provides new and exciting possibilities, it is but one of the many opportunities direct surface analysis offers the lipid researcher. In this review we describe the current state-of-the-art in the direct surface analysis of lipids with a focus on tissue sections, intact cells and thin-layer chromatography substrates.The suitability of these different approaches towards analysis of the major lipid classes along with their current and potential applications in the field of lip...

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“…The specifications of the infrared matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometer (IR-MALDI-o-TOF-MS; MDS Sciex, Concord) have been described in two recent publications (14,27). Direct TLC-IR-MALDI-o-TOF-MS analysis was done in situ from immunopositive bands (see TLC Overlay Assay) as described in detail by Distler et al (14).…”

Section: Infrared Matrix-assisted Laser Desorption/ionization Orthogomentioning

confidence: 99%

Tumor-associated CD75s- and iso-CD75s-gangliosides are potential targets for adjuvant therapy in pancreatic cancer

Distler

Souady²,

Hülsewig³

et al. 2008

Molecular Cancer Therapeutics

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Pancreatic adenocarcinoma confers one of the highest mortality rates in malignant human tumors with very poor prognosis. Because as yet no treatments are available that produce a substantial survival benefit for this fatal neoplasia, new therapeutic concepts are urgently required to support cancer standard treatment. In search of tumorassociated gangliosides with therapeutic background, we probed a random collection of cancerous and adjacent normal postoperative tissue samples from 38 patients for the expression of CD75s-and iso-CD75s-gangliosides. We exhaustively analyzed the expression of CD75s-1-ganglioside (IV 6 Neu5Ac-nLc4Cer) and structurally closely related iso-CD75s-1-ganglioside (IV 3 Neu5Ac-nLc4Cer) by means of immunohistology of cryosections and semiquantitative TLC of tissue lipid extracts combined with mass spectrometry. CD75s-1-and iso-CD75s-1-ganglioside showed an elevated expression in 42% and 66% of the tumors, respectively, indicating a significant association with neoplastic transformation (P = 0.001). Thus, increased expression of CD75s-1-and iso-CD75s-1-gangliosides renders these cell surface molecules promising candidates for oncologic applications. Further statistical analysis revealed a significant enhancement of CD75s-1-ganglioside in the group of less differentiated tumors (grade >2) suggesting this ganglioside as a potential marker for poor differentiation. The CD75s-specific antitumor drug rViscumin, which represents the recombinant counterpart of the ribosome-inactivating lectin viscumin, has successfully passed clinical phase I trials and provides an opportunity for treating pancreatic cancer. Consequently, if an enhanced expression is existent in malignant tissues, we propose the targeting of CD75s-gangliosides with rViscumin as a novel potential strategy in adjuvant treatment of pancreatic malignancies. [Mol Cancer Ther 2008;7(8):2464 -75]

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Analysis of Gangliosides Directly from Thin-Layer Chromatography Plates by Infrared Matrix-Assisted Laser Desorption/Ionization Orthogonal Time-of-Flight Mass Spectrometry with a Glycerol Matrix

Cited by 119 publications

References 41 publications

Ganglioside analysis by thin-layer chromatography matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry

Ganglioside analysis by thin-layer chromatography matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry

Surface analysis of lipids by mass spectrometry: More than just imaging

Tumor-associated CD75s- and iso-CD75s-gangliosides are potential targets for adjuvant therapy in pancreatic cancer

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