Quantitative evaluation of sphingolipids using delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry with sphingosylphosphorylcholine as an internal standard
| Autor: | Hironori Kobayashi, Nobuyuki Takahashi, Seiji Yamaguchi, Masaru Tasaka, Takehisa Fujiwaki, Toshio Shimada, Yo Murakami |
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| Rok vydání: | 2006 |
| Předmět: |
Delayed extraction
Chromatography Clinical Biochemistry Globotriaosylceramide Cell Biology General Medicine medicine.disease Mass spectrometry Biochemistry Sphingolipid Fabry disease Analytical Chemistry chemistry.chemical_compound Matrix-assisted laser desorption/ionization Glycolipid chemistry medicine lipids (amino acids peptides and proteins) Quantitative analysis (chemistry) |
| Zdroj: | Journal of Chromatography B. 832:97-102 |
| ISSN: | 1570-0232 |
| DOI: | 10.1016/j.jchromb.2005.12.039 |
| Popis: | Fabry disease is a glycolipid storage disorder caused by a defect of alpha-galactosidase A, and characterized by the systemic deposition of glycosphingolipids with terminal alpha-galactosyl moieties, mainly globotriaosylceramide, in tissues. Using delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry (DE MALDI-TOF-MS), we analyzed the sphingolipids in the cardiac valves from a 49-year-old male patient with Fabry disease who suffered from congestive cardiac failure. Crude lipids were extracted from the cardiac valves with chloroform and methanol. After mild alkaline treatment of the crude lipids, a sphingolipid fraction was prepared and analyzed by DE MALDI-TOF-MS. The results were as follows: (a) ion peaks with m/z values corresponding to different ceramide trihexoside (CTH) species were detected; (b) with sphingosylphosphorylcholine (SPC) as the internal standard for semi-quantification of CTH, the relative peak height of CTH was calculated and plotted versus its amount loaded on the sample plate for MALDI-TOF-MS. The relative peak height of CTH with fatty acid C16:0 showed linearity between 0 and 50 ng CTH (regression coefficient, r>0.95); (c) semi-quantitative analysis revealed striking accumulation of CTH in the cardiac valves from the patient with Fabry disease. It was indicated that the accumulation of CTH in cardiac valves from Fabry disease patients can be detected with the DE MALDI-TOF-MS method. SPC is commercially available, and this semi-quantitative method involving MALDI-TOF-MS was found to be convenient, reliable and useful for CTH. It is expected to be applied to the quantification of CTH in small amounts of body fluids or other tissues and to clinical examination. It is also expected to be applicable to the quantification of other glycosphingolipids. |
| Databáze: | OpenAIRE |
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