Acid sphingomyelinase deficiency (ASMd, Niemann-Pick disease A/B) and Niemann-Pick type C disease (NPC) share core clinical symptoms. Initial diagnostic discrimination of these two rare lysosomal storage diseases is thus difficult. As sphingomyelin accumulates in ASMd as well as NPC, lysosphingomyelin (sphingosylphosphorylcholine) and its m/z 509 analog were suggested as biomarkers for both diseases. Herein we present results of simultaneous LC-ESI-MS/MS measurements of lysosphingomyelin and lysosphingomyelin 509 in plasma and dried blood spots (DBS) collected from ASMd and NPC patients and suggest that the plasma but not DBS levels of the two analytes allow differential biochemical screening of ASMd and NPC.
- MeSH
- biologické markery krev MeSH
- chromatografie kapalinová metody MeSH
- fosforylcholin analogy a deriváty krev MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- lidé MeSH
- Niemannova-Pickova nemoc typu A krev diagnóza MeSH
- Niemannova-Pickova nemoc typu B krev diagnóza MeSH
- Niemannova-Pickova nemoc typu C krev diagnóza MeSH
- sfingosin analogy a deriváty krev MeSH
- studie případů a kontrol MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- test suché kapky krve metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
In recent years, mass spectrometry (MS) has become the dominant technology in lipidomic analysis. It is widely used in diagnosis and research of lipid metabolism disorders including those characterized by impairment of lysosomal functions and storage of nondegraded-degraded substrates. These rare diseases, which include sphingolipidoses, have severe and often fatal clinical consequences. Modern MS methods have contributed significantly to achieve a definitive diagnosis, which is essential in clinical practice to begin properly targeted patient care. Here we summarize MS and tandem MS methods used for qualitative and quantitative analysis of sphingolipids (SL) relative to the diagnostic process for sphingolipidoses and studies focusing on alterations in cell functions due to these disorders. This review covers the following topics: Tandem MS is sensitive and robust in determining the composition of sphingolipid classes in various biological materials. Its ability to establish SL metabolomic profiles using MS bench-top analyzers, significantly benefits the first stages of a diagnosis as well as metabolic studies of these disorders. It can thus contribute to a better understanding of the biological significance of SL.
- MeSH
- lidé MeSH
- sfingolipidózy diagnóza MeSH
- sfingolipidy analýza chemie fyziologie MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Sphingolipid ceramide N-deacylase (SCDase, EC 3.5.1.69) is a hydrolytic enzyme isolated from Pseudomonas sp. TK 4. In addition to its primary deacylation function, this enzyme is able to reacylate lyso-sphingolipids under specific conditions. We immobilised this enzyme on magnetic macroporous cellulose and used it to semisynthesise C17:0 glucosylceramide and C17:0 sulphatide, which are required internal standards for quantification of the corresponding glycosphingolipids (GSL) by tandem mass spectrometry. A high rate of conversion was achieved for both lipids (80% for C17:0 sulphatide and 90% for C17:0 glucosylceramide). In contrast to synthesis with a soluble form of the enzyme, use of immobilised SCDase significantly reduced the contamination of the sphingolipid products with other isoforms, so further purification was not necessary. Our method can be effectively used for the simple preparation of specifically labelled sphingolipids of high isoform purity for application in mass spectrometry.
- MeSH
- amidohydrolasy chemie MeSH
- bakteriální proteiny chemie MeSH
- enzymy imobilizované chemie MeSH
- glukosylceramidy chemická syntéza chemie MeSH
- glykosfingolipidy analýza MeSH
- hmotnostní spektrometrie metody normy MeSH
- hydrolýza MeSH
- Pseudomonas enzymologie MeSH
- referenční standardy MeSH
- stereoizomerie MeSH
- sulfoglykosfingolipidy chemická syntéza chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- alfa-glukosidasy chemie MeSH
- isoelektrická fokusace MeSH
- krysa rodu rattus MeSH
- mikroklky enzymologie MeSH
- morčata MeSH
- neuraminidasa metabolismus MeSH
- rozpustnost MeSH
- techniky in vitro MeSH
- tenké střevo enzymologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- morčata MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- barvicí látky MeSH
- biotransformace MeSH
- diazoniové sloučeniny metabolismus MeSH
- jaterní mikrozomy metabolismus MeSH
- krysa rodu rattus MeSH
- nádorová transformace buněk MeSH
- naftoly metabolismus MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- MeSH
- biotransformace MeSH
- chromatografie na tenké vrstvě MeSH
- cytochromy MeSH
- diazoniové sloučeniny analýza MeSH
- flavony MeSH
- jaterní mikrozomy metabolismus MeSH
- krysa rodu rattus MeSH
- nádorová transformace buněk analýza MeSH
- naftoly metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
