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Direct matrix-assisted laser desorption/ionization mass spectrometric analysis of glycosphingolipids on thin layer chromatographic plates and transfer membranes.
Rapid Commun Mass Spectrom. 1999; 13(18):1838-49.RC

Abstract

Results are reported for analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) of native glycosphingolipids (GSLs) after development on thin layer chromatographic plates and after heat transfer of the GSLs from the plates to several types of polymer membranes. The spectral quality is better for membrane-bound analytes, in terms of sensitivity, mass resolution and background interference. The sensitivity gain compared with liquid secondary ion mass spectrometry (LSIMS) of GSLs on thin layer plates is 1-2 orders of magnitude (detection limits of 5-50 pmol vs. 1-10 nmol). Resolution and mass accuracy (0.1%) are limited by the irregular membrane surfaces and this effect cannot be entirely compensated by delayed extraction. The best results were obtained with a polyvinylidene difluoride (PVDF) P membrane, with irradiation from a nitrogen laser. Although the Nafion membrane could not be used for molecular weight profiling, its acidic character led to sample hydrolysis at the glycosidic linkages, thus yielding a series of fragments that could be used to determine the sequence of carbohydrate residues. Structural information could also be obtained by post-source decay (PSD) experiments on mass-selected precursor ions. Samples containing both neutral and acidic components were characterized in a 1:1 combination of 2, 5-dihydroxybenzoic acid and 2-amino-5-nitropyridine. GSLs that exhibited binding to antibodies in an overlay assay on the TLC plate were transferred to membranes and analyzed by MALDI-TOFMS without interference from the antibody or the salts and buffers used during the binding and visualization steps. Taking advantage of the insights into sample preparation gained from these studies, future research will extend this approach to analysis by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) with an external ion source.

Authors+Show Affiliations

Guittard J
Mass Spectrometry Resource, Departments of Biochemistry and Biophysics, Boston University School of Medicine, Boston, MA 02118-2526, USA.
Hronowski XL
No affiliation info available
Costello CE
No affiliation info available

MeSH

AnimalsChromatography, Thin LayerGlycosphingolipidsHumansSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10482898

Citation

Guittard, J, et al. "Direct Matrix-assisted Laser Desorption/ionization Mass Spectrometric Analysis of Glycosphingolipids On Thin Layer Chromatographic Plates and Transfer Membranes." Rapid Communications in Mass Spectrometry : RCM, vol. 13, no. 18, 1999, pp. 1838-49.
Guittard J, Hronowski XL, Costello CE. Direct matrix-assisted laser desorption/ionization mass spectrometric analysis of glycosphingolipids on thin layer chromatographic plates and transfer membranes. Rapid Commun Mass Spectrom. 1999;13(18):1838-49.
Guittard, J., Hronowski, X. L., & Costello, C. E. (1999). Direct matrix-assisted laser desorption/ionization mass spectrometric analysis of glycosphingolipids on thin layer chromatographic plates and transfer membranes. Rapid Communications in Mass Spectrometry : RCM, 13(18), 1838-49.
Guittard J, Hronowski XL, Costello CE. Direct Matrix-assisted Laser Desorption/ionization Mass Spectrometric Analysis of Glycosphingolipids On Thin Layer Chromatographic Plates and Transfer Membranes. Rapid Commun Mass Spectrom. 1999;13(18):1838-49. PubMed PMID: 10482898.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Direct matrix-assisted laser desorption/ionization mass spectrometric analysis of glycosphingolipids on thin layer chromatographic plates and transfer membranes. AU - Guittard,J, AU - Hronowski,X L, AU - Costello,C E, PY - 1999/9/14/pubmed PY - 1999/9/14/medline PY - 1999/9/14/entrez SP - 1838 EP - 49 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 13 IS - 18 N2 - Results are reported for analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) of native glycosphingolipids (GSLs) after development on thin layer chromatographic plates and after heat transfer of the GSLs from the plates to several types of polymer membranes. The spectral quality is better for membrane-bound analytes, in terms of sensitivity, mass resolution and background interference. The sensitivity gain compared with liquid secondary ion mass spectrometry (LSIMS) of GSLs on thin layer plates is 1-2 orders of magnitude (detection limits of 5-50 pmol vs. 1-10 nmol). Resolution and mass accuracy (0.1%) are limited by the irregular membrane surfaces and this effect cannot be entirely compensated by delayed extraction. The best results were obtained with a polyvinylidene difluoride (PVDF) P membrane, with irradiation from a nitrogen laser. Although the Nafion membrane could not be used for molecular weight profiling, its acidic character led to sample hydrolysis at the glycosidic linkages, thus yielding a series of fragments that could be used to determine the sequence of carbohydrate residues. Structural information could also be obtained by post-source decay (PSD) experiments on mass-selected precursor ions. Samples containing both neutral and acidic components were characterized in a 1:1 combination of 2, 5-dihydroxybenzoic acid and 2-amino-5-nitropyridine. GSLs that exhibited binding to antibodies in an overlay assay on the TLC plate were transferred to membranes and analyzed by MALDI-TOFMS without interference from the antibody or the salts and buffers used during the binding and visualization steps. Taking advantage of the insights into sample preparation gained from these studies, future research will extend this approach to analysis by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) with an external ion source. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/10482898/Direct_matrix_assisted_laser_desorption/ionization_mass_spectrometric_analysis_of_glycosphingolipids_on_thin_layer_chromatographic_plates_and_transfer_membranes_ L2 - https://doi.org/10.1002/(SICI)1097-0231(19990930)13:18<1838::AID-RCM726>3.0.CO;2-9 DB - PRIME DP - Unbound Medicine ER -
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