Vydání první 128 stran : ilustrace ; 30 cm
Vysokoškolská učebnice, která se zaměřuje na obecnou biochemii, klinickou biochemii a pathobiochemii.
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- biochemie
- NLK Publikační typ
- učebnice vysokých škol
The determination of α-keto acids derived from amino acids is currently the most reliable approach for the diagnosis of some congenital metabolic diseases. An HPLC method for the simultaneous measurement of selected α-keto acids in dried blood samples has been developed and evaluated. Blood spot samples from a group of healthy blood donors were collected onto #903 Specimen Collection Paper. Prior the separation, the α-keto acids were derivatized with 1,2-diamino-4,5-dimethoxybenzene to the corresponding 3-substituted-6,7-dimethoxy-2(1 H)-quinoxalinol derivatives. For the separation, a reverse-phase column LichroCart 125-4, Purospher RP-18e, 5 μm, was used. The mixture of 25% ACN in deionized water (mobile phase A) and 100% ACN (mobile phase B) were used for a gradient elution of α-keto acids derivatives. Analytical performance of this method is satisfactory for all α-keto acids. The intra-assay and inter-assay coefficients were below 10% and recoveries were close to 100%. We have developed relatively simple, rapid, selective and sufficiently sensitive HPLC method with fluorescence detection for the determination of selected α-keto acids in dried blood samples. The presented method is suitable for clinical testing purposes.
A method is described for the determination of fatty acids in dried sweat spot and plasma samples using gas chromatography with flame ionization detection. Plasma and dried sweat spot samples were obtained from a group of blood donors. The sweat was collected from each volunteer during exercise. Sweat was spotted onto collection paper containing butylated hydroxytoluene. Fatty acids were derivatized with acetyl chloride in methanol to form methyl esters of fatty acids. The fatty acids in dried sweat spot samples treated with butylated hydroxytoluene and stored at -20°C were stable for 3 months. Our results indicate that sweat contains, among fatty acids with short chain, also fatty acids with long chain and unsaturated fatty acids. Linear relationships between percentage content of selected fatty acids in dried sweat spot and plasma were observed.
- MeSH
- chromatografie plynová * MeSH
- estery analýza MeSH
- lidé MeSH
- mastné kyseliny analýza MeSH
- plamínková ionizace * MeSH
- pot chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: The determination of amino acids can be a reliable approach for extended diagnosis of liver diseases. This is because liver disease can be a cause of impaired amino acid metabolism. Therefore, a method for the determination of serum amino acids, applicable for clinical purposes, is necessary. OBJECTIVES: The aim of this study was to find differences in the levels of selected amino acids between patients with liver disease and a control group. MATERIAL AND METHODS: Samples of peripheral venous blood were obtained from a group of patients with liver disease (n = 131, 59 women at an average age of 60 years and 72 men at an average age of 52 years) and a control group (n = 105, 47 women at an average age of 62 years and 58 men at an average age of 58 years). Before the separation, the amino acids were derivatized with naphthalene-2,3-dicarboxaldehyde. For the separation, reverse phase column was used. The effluent was monitored with a fluorescence detector. RESULTS: There were significant differences in the concentrations of some amino acids between the patients and the control group, but also between women and men. Correlations between some amino acids and markers of liver blood tests and lipid metabolism were observed. CONCLUSIONS: A simple, relatively rapid and selective HPLC method with fluorescence detection for the determination of selected amino acids in serum has been developed.
- MeSH
- aminokyseliny krev MeSH
- analýza rozptylu MeSH
- fluorescence MeSH
- kalibrace MeSH
- lidé středního věku MeSH
- lidé MeSH
- limita detekce MeSH
- nemoci jater krev diagnóza MeSH
- neparametrická statistika MeSH
- studie případů a kontrol MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Oxidative stress is an imbalance between reactive oxygen species exposure and the ability of organisms to detoxify the reactive intermediates and to repair the oxidative damage of biologically important molecules. Many clinical studies of oxidative stress unfortunately provide conflicting and contradictory results. The ability of antioxidant systems to adequately respond to oxidative stress can be used in laboratory diagnostics. In the present review, methods using the ratio of reduced and oxidized forms of uric acid, ascorbic acid, glutathione and coenzyme Q10 as suitable indicators of oxidative stress are discussed. From the mentioned publications it is evident that suitable sample preparation prior to analysis is crucial.
- MeSH
- antioxidancia chemie MeSH
- biologické markery chemie MeSH
- lidé MeSH
- oxidace-redukce MeSH
- oxidační stres * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
In the present review, methods for the determination of glutathione and glutathione disulfide are discussed. The main topic of this review is the suitable sample preparation prior to analysis which is absolutely crucial. The role of glutathione as a marker of oxidative stress is small; however, the ability of glutathione to adequately respond to oxidative stress by increasing the content of its oxidized form, especially glutathione disulfide, is meaningful. Therefore, methods that can properly quantify both forms of glutathione are essential to monitor the oxidative stress in humans.
A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.
- MeSH
- dospělí MeSH
- glutathion krev chemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- limita detekce MeSH
- lineární modely MeSH
- methanol MeSH
- mladiství MeSH
- mladý dospělý MeSH
- reprodukovatelnost výsledků MeSH
- test suché kapky krve metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Vydání první 128 stran : ilustrace ; 30 cm
- MeSH
- aminokyseliny metabolismus MeSH
- biochemie MeSH
- hem metabolismus MeSH
- metabolismus lipidů MeSH
- metabolismus sacharidů MeSH
- nukleotidy metabolismus MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- biochemie
- NLK Publikační typ
- učebnice vysokých škol
Uric acid is the final product of human purine metabolism. It was pointed out that this compound acts as an antioxidant and is able to react with reactive oxygen species forming allantoin. Therefore, the measurement of allantoin levels may be used for the determination of oxidative stress in humans. The aim of the study was to clarify the antioxidant effect of uric acid during intense exercise. Whole blood samples were obtained from a group of healthy subjects. Allantoin, uric acid, and malondialdehyde levels in plasma and erythrocytes were measured using a HPLC with UV/Vis detection. Statistical significant differences in allantoin and uric acid levels during short-term intense exercise were found. Immediately after intense exercise, the plasma allantoin levels increased on the average of 200 % in comparison to baseline. Plasma uric acid levels increased slowly, at an average of 20 %. On the other hand, there were no significant changes in plasma malondialdehyde. The results suggest that uric acid, important antioxidant, is probably oxidized by reactive oxygen species to allantoin. Therefore allantoin may be suitable candidate for a marker of acute oxidative stress.
- MeSH
- alantoin krev MeSH
- běh fyziologie MeSH
- biologické markery krev MeSH
- dospělí MeSH
- erytrocyty metabolismus MeSH
- hemoglobiny analýza MeSH
- kyselina mléčná krev MeSH
- kyselina močová krev MeSH
- lidé MeSH
- malondialdehyd krev MeSH
- mladý dospělý MeSH
- reaktivní formy kyslíku metabolismus MeSH
- zátěžový test MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We describe a relatively simple method for the determination of glutathione (GSH) and glutathione disulfide (GSSG) in human whole blood. We have used an HPLC with coulometric electrochemical detection for the simultaneous measurement of GSH and GSSG. Diluted and filtered trichloroacetic acid extracts were injected directly into the HPLC system and were eluted isocratically on a Polaris 5u C18-A, 250 × 4.6 mm analytical column. Glutathione in samples extracted with trichloroacetic acid and diluted with 1.0 mM hydrochloric acid was stable at 4 °C for at least 8 h. The analytical performance of this method is satisfactory: the intra-assay and inter-assay coefficients of variation were below 10%. Quantitative recoveries from spiked whole blood samples were at intervals 91.6–97.6% for GSH and 85.0–104.4% for GSSG. The linear range is 5.0–2000.0 µmol/l, with a detection limit of 2.1 µmol/l (signal-to-noise ratio = 3) for GSH and 2.0–250.0 µmol/l, with a detection limit of 0.9 µmol/l for GSSG.
