Metabolomics has become an important tool in clinical research and diagnosis of human diseases. In this work we focused on the diagnosis of inherited metabolic disorders (IMDs) in plasma samples using a targeted metabolomic approach. The plasma samples were analyzed with the flow injection analysis method. All the experiments were performed on a QTRAP 5500 tandem mass spectrometer (AB SCIEX, U.S.A.) with electrospray ionization. The compounds were measured in a multiple reaction monitoring mode. We analyzed 50 control samples and 34 samples with defects in amino acid metabolism (phenylketonuria, maple syrup urine disease, tyrosinemia I, argininemia, homocystinuria, carbamoyl phosphate synthetase deficiency, ornithine transcarbamylase deficiency, nonketotic hyperglycinemia), organic acidurias (methylmalonic aciduria, propionic aciduria, glutaric aciduria I, 3-hydroxy-3-methylglutaric aciduria, isovaleric aciduria), and mitochondrial defects (medium-chain acyl-coenzyme A dehydrogenase deficiency, carnitine palmitoyltransferase II deficiency). The controls were distinguished from the patient samples by principal component analysis and hierarchical clustering. Approximately 80% of patients were clearly detected by absolute metabolite concentrations, the sum of variance for first two principle components was in the range of 44-55%. Other patient samples were assigned due to the characteristic ratio of metabolites (the sum of variance for first two principle components 77 and 83%). This study has revealed that targeted metabolomic tools with automated and unsupervised processing can be applied for the diagnosis of various IMDs.
- MeSH
- analýza hlavních komponent MeSH
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- metabolom MeSH
- metabolomika metody MeSH
- mladiství MeSH
- předškolní dítě MeSH
- průtoková injekční analýza MeSH
- reprodukovatelnost výsledků MeSH
- shluková analýza MeSH
- tandemová hmotnostní spektrometrie MeSH
- vrozené poruchy metabolismu aminokyselin krev diagnóza MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nucleotides play an important role in human metabolism and their disorders often result in severe health impairment. Separation of major cellular nucleotides from dry blood spots was achieved by capillary electrophoresis. On the basis of incubations of erythrocytes with nucleotides, possible applications of the method in screening tests are proposed. Deproteinated dry blood spot extracts were analysed in a silica capillary using UV detection. The separation buffer was a solution of citric acid and cetyltrimethylammonium bromide, its pH was adjusted to 4.3 with 4-aminobutanoic acid (GABA). The overall nucleotide profile of blood spot extracts is comparable with that of fresh blood, although their contents may slightly change during drying and long-term storage of the spots. The method offers reproducibility within 5 %, accuracy below 14 %, recovery of total adenine nucleotides 89.8±10.5 % and the limit of detection 1.2 ?mol L?1 (R2 > 0.99) for uridine triphosphate and inosine monophosphate. Nucleotide profiles simulating disorders of purine metabolism were obtained by incubation of erythrocytes with nucleosides. The use of dry blood spots instead of fresh blood facilitates collection, transport and storage of blood samples and is patient-friendly. This method may be applicable to screening of inherited metabolic disorders.
- MeSH
- analýza potravin * metody normy MeSH
- chléb analýza MeSH
- cukrovinky analýza MeSH
- dietní tuky nenasycené analýza MeSH
- hodnocení stavu výživy MeSH
- kontrola potravin * metody normy MeSH
- lidé MeSH
- margarín analýza MeSH
- máslo analýza MeSH
- mastné kyseliny analýza klasifikace MeSH
- zmrzlina analýza MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
