Gyula Wittmann

Determination of glutathione and glutathione disulfide in biological samples: An in-depth review

Glutathione (GSH) is a thiol-containing tripeptide, which plays central roles in the defence against oxidative damage and in signaling pathways. Upon oxidation, GSH is transformed to glutathione disulfide (GSSG). The concentrations of GSH and GSSG and their molar ratio are indicators of cell functionality and oxidative stress. Assessment of redox homeostasis in various clinical states and medical applications for restoration of the glutathione status are of growing importance. This review is intended to provide a state-of-the-art overview of issues relating to sample pretreatment and choices for the separation and detection of GSH and GSSG. High-performance liquid chromatography, capillary electrophoresis and gas chromatography (as techniques with a separation step) with photometric, fluorimetric, electrochemical and mass spectrometric detection are discussed, stress being laid on novel approaches.

Analysis of volatile organic compounds using gas chromatography.

The focus of this review is the analysis of volatile organic compounds (VOCs) by gas chromatography (GC) in the field of environmental, food, flavour and fragrance, medical and forensic sciences. New trends in sample injection, separation and detection are covered, including multi-dimensional and high-speed GC. Attention is drawn to a growing interest in quality assessment. From the review, it is clear that it remains a challenge to generate multi-component gaseous standards of VOCs at ppbv and pptv.

Newborn Screening for Lysosomal Storage Disorders in Hungary

Even though lysosomal storage disorders (LSDs) are considered to be orphan diseases, they pose a highly relevant cause for morbidity and mortality as their cumulative prevalence is estimated to be 1:4,000. This is especially important as treatment in form of enzyme replacement therapy, substrate reduction therapy or stem cell transplantation is amenable for some LSDs. It is plausible that an early start of treatment might improve the overall prognosis and, even more important, prevent irreversible damage of key organs. To get a more precise insight into the real frequency of some LSDs in the general population, we screened 40,024 samples from the Hungarian newborn screening (NBS) program in Szeged for Fabry disease (FD), Gaucher disease (GD), Pompe disease (PD), and Niemann-Pick A/B (NPB) disease using tandem mass spectrometry. Altogether, 663 samples (1.66%) were submitted for retesting. Genetic confirmation was carried out for 120 samples with abnormal screening results after retesting, which identified three cases of GD, three cases of FD, nine cases of PD, and two cases with NPB. In some cases, we detected up to now unknown mutations - one in NPB and seven in PD - which raise questions about the clinical consequences of a NBS in the sense of late-onset manifestations. Overall, we conclude that screening for LSDs by tandem MS/MS followed by a genetic workup in identified patients is a robust, easy, valid, and feasible technology in newborn screening programs. Furthermore, early diagnosis of LSDs gives a chance to early treatment, but needs more clinical long-term data especially regarding the consequence of private mutations.

Homogeneous and heterogeneous catalytic asymmetric reactions. Part III. New investigations concerning the preparation of reproducible MRNi catalyst

Abstract A new method is described for the preparation of Raney Ni catalyst modified with R,R-tartaric acid (MRNi). All experimental conditions which may have a significant impact on the preparation of a reproducible enantioselective catalyst were studied. Experiments were also performed to permit a comparison with literature data.

Homogeneous and heterogeneous catalytic asymmetric reactions IV. Hydrogenation of the Ni complexes of cinnamic acid salts over MRNi catalyst

Abstract Substituted cinnamic acids containing a prochiral CC bond, and their Li I , K I , Ni II and Ca II salts, were prepared and their surface complexes on Ni were studied over a Raney-Ni catalyst modified with tartaric acid. The hydrogenation product of sodium cinnamate is optically active. Of the alkali metal salts of ( E )-α-phenylcinnamic acid, the hydrogenation of the Na salt gives the highest optical yield (17%). The solvent used significantly influences the selectivity. The published data for the chiral RH complexes are listed for comparison with the results of the homogeneous and heterogeneous hydrogenations described. The enantioselectivity in the presence of Raney-Ni is the result of a complex formed by the interaction of ( R,R )-tartaric acid with the substrate.

Comparison of ozone-based and other (VUV and TiO2/UV) radical generation methods in phenol decomposition

Abstract The degradation reaction kinetics and chemical mechanism of phenol decomposition by ozonation, TiO2-photocatalysis and vacuum ultraviolet (VUV) photolysis were investigated. The concentration dependences of the aromatic and aliphatic intermediates and the TOC content were compared as a function of the phenol conversion. The concentration profiles and the TOC curves obtained with each method were very similar. The results suggest a similar chemical mechanism for the transformation of phenol irrespective of the starting active component (a positive hole on the surface of the catalysts, a VUV photon, ozone or the hydroxyl radical).

UV-induced decomposition of ozone and hydrogen peroxide in the aqueous phase at pH 2-7

Abstract The photolysis of ozone and formation of hydrogen peroxide were investigated in solution of pH 2–7, in a 200 cm3 photoreactor in the incident photon flow range 9.6 x 10−8 - 4.2 x 10−7 einstein s−1. The quantum yield of the primary photochemical reactions was measured in a direct way by suppressing the secondary radical reactions. The determined quantum yields of the photo-decompositions of ozone and hydrogen peroxide were 0.42 ± 0.042 ± 0.04 and 0.49 ± 0.04, respectively. A correct mathematical treatment is given for calculation of the light absorption of the individual components of a multi-absorbent reaction mixture. On the basis of the literature data and die present results, a probable chemical and reaction kinetic model was proposed to characterize the investigated reaction systems. Reaction kinetic simulations demonstrated that the model predicts a good fit to the measured data with the preferred literature rate constants, except that for the HO3 radical decomposition reaction. A reasonable reduction of this rate coefficient value is in accordance with the latest published results.

Trace analysis of trichlorobenzenes in fish by microwave-assisted extraction and gas chromatography-electron-capture detection

An analytical method consisting of extraction, clean-up, and analysis by gas chromatography-electron-capture detection (GC-ECD) was developed for the determination of trichlorobenzenes (TCBs) in fish samples. Two extraction methods, saponification and liquid-liquid extraction (S-LLE), and microwave-assisted extraction (MAE), were evaluated. In both cases, n-pentane was used as the extraction solvent. For S-LLE, the recoveries ranged from 66.6+/-9.1% for 1-bromo-4-chlorobenzene (4-BCB) to 93.5+/-4.9% for 1,2,4-trichlorobenzene (1,2,4-TCB). The recoveries were significantly lower, between 31.0+/-3.9% for 1,2,3-trichlorobenzene (1,2,3-TCB) and 52.3+/-3.0% for 1,3,5-trichlorobenzene (1,3,5-TCB), in the absence of fish. Proteins and glycerides of the fish tissue seemed to compete with TCBs for the base, and hence decreased their decomposition rate. In the case of MAE, the recoveries were highly dependent on the pressure applied during extraction. At 5 bar, much higher recoveries were obtained, from 66.7+/-15.6% for 4-BCB to 79.9+/-13.6% for 1,2,4-TCB, than at 1 bar. Sulfur formation was, however, observed at 5 bar, and interfered with the GC-ECD analysis of TCBs. Sulfur was adequately removed by copper powder treatment, which was shown not to affect the recovery of analytes. The recoveries of target analytes by S-LLE and MAE did not differ statistically (t-test, alpha = 0.01). Both methods were appropriate for the detection of TCBs at concentration levels typically observed in marine biota, i.e. approximately 1 ng/g. S-LLE was, however, more time consuming, and required larger volumes of high-purity organic solvents than MAE.

Enhanced methylglyoxal formation in the erythrocytes of hemodialyzed patients

Methylglyoxal (MG) contributes significantly to the carbonyl stress in uremia; however, the reason for its increased concentration is not clear. Thus, the present study was aimed to investigate the formation and degradation of MG in the erythrocytes of hemodialyzed (HD) patients with end-stage renal disease. In 22 nondiabetic patients on long-term HD, erythrocyte MG and d-lactate levels, glyoxalase activities, and whole blood reduced glutathione content were determined. The data were compared with those from 22 healthy controls. Erythrocyte MG and d-lactate production were also investigated in vitro under normoglycemic (5 mmol/L) and hyperglycemic (50 mmol/L) conditions. The erythrocyte MG levels were elevated (P < .001) in the HD patients. The blood reduced glutathione content and glyoxalase I activity were similar to the control levels, but the glyoxalase II activity was significantly (P < .005) increased. In the normoglycemic in vitro model, production of both MG (P < .001) and d-lactate (P < .002) was significantly enhanced in the HD erythrocytes relative to the controls. During hyperglycemia, the MG formation and degradation rates were further increased (P < .001). The present study demonstrated an increased formation of MG in the erythrocytes of HD patients. This seemed to be related to a glucose metabolism disturbance of the cells. The degradation system of MG was also activated; still, it was not able to counteract the high rate of MG formation. The alterations and imbalance of these metabolic processes may contribute to the carbonyl overload and stress in the HD patients.

Simultaneous Determination of Hydrogen Peroxide and Organic Hydroperoxides in Water

The kinetics of the reactions of H 2 O 2 and of methyl, ethyl, tert -butyl, and cumene hydroperoxides with I m were investigated in the presence and absence of molybdate as catalyst. These results were utilized to develop an analytical method for the simultaneous determination of H 2 O 2 and organic hydroperoxides in aqueous solutions. The total amount of H 2 O 2 and organic hydroperoxides can be determined by the spectrophotometric measurement of