M. Bartoszek

The spectroscopic studies of humic acid extracted from sediment collected at different seasons

Goczałkowice Reservoir is the biggest water reservoir in the south of Poland. For our studies bottom sediments were collected from eight different places of the reservoir at various seasons of the year. EPR spectroscopy was applied to analyse both quantitatively and qualitatively the humic acids (HA) present in the samples. EPR spectra of the extracted HA exhibited broad lines from the paramagnetic metal ions and narrow lines from free radicals. The values of the free radical concentration obtained for HA amounted to 1.14-13.6 × 1016 spin g(-1) depending on the season and the place of sample collection. The values of the g factor obtained for HA were of the range 2.0027-2.0035. The EPR studies show that HA extracted from bottom sediment collected at various points of the Goczałkowice Reservoir exhibit similar physical-chemical properties. It was also observed that the depth of the reservoir affects the content of the oxygen functional groups as well as the free radical concentration in HA. The identification of the functional groups was done by means of IR. UV/VIS spectroscopy was used to estimate the maturity of the HA.

NMR study of the humification process during sewage sludge treatment

Humic acids, extracted from sludge at the biologic-mechanical sewage treatment plant in Jastrzebie Zdroj, have been investigated by means of (1)H, (13)C and (31)P NMR spectroscopy. Sludge samples for studies were taken from the primary settling tank, the nitrification chamber, the digestion chamber and the sludge drying beds. The (1)H NMR analysis of humic acid extracted from sludge at various stages of sewage treatment confirmed the presence of the functional groups that are characteristic for humic substances, and the analysis showed changes in their relative intensities. The (13)C NMR indicated that the aromatisation of the humic acid increased during sewage treatment. Moreover, the analysis of the (31)P NMR spectra allowed us to observe the changes in the phosphorus groups of the studied humic acids.

A study of the antioxidant properties of beers using electron paramagnetic resonance

The antioxidant properties of various kinds of beers were investigated using electron paramagnetic resonance (EPR) spectroscopy. This was possible by measuring the changes in the intensity of the EPR spectrum that resulted from the interaction of the stable radical DPPH (1,1-diphenyl-2-picrylhydrazyl) with the antioxidants found in a beer sample. The antioxidant capacity was then presented in Trolox Equivalents, e.g. μM trolox in a beer sample of 100ml. The influence of the type, colour, the content of the extract and alcohol on the antioxidant activities of commercial beer samples was investigated using two-way hierarchical clustering and analysis of variance. The results showed that all of the beers investigated exhibit antioxidant properties. By performing an analysis of variance, it was found that the value of the antioxidant capacity significantly (0.05 level of significance) depends on the content of the extract and the colour of the beer. It seems that additives also influence the antioxidant properties to some extent, but neither the alcohol content nor the kind of fermentation affects the antioxidant properties of beer.

A study of magnetic anisotropy of blood cells

Abstract This paper presents an experimental study of the magnetic anisotropy of blood cells in magnetic field up to 1.8 T and in temperature range of 75 – 295 K by means of torque method. Magnetic field can induce a marked uniaxial anisotropy. The effective anisotropy constant K 1eff does not depend on temperature. Magnetic-induced anisotropy of investigated samples cannot be explained by diamagnetic shape anisotropy of individual blood cells. A tendency to aggregate into chains and small groups of hemoglobin particles and conversion into methemoglobin with high spin Fe 3+ -ion is considered.

Antioxidant Activity of Selected Thyme (Thymus L.) Species and Study of the Equivalence of Different Measuring Methodologies.

This study presents the results of comparative evaluation of the antioxidant activity of the phenolic fraction exhaustively extracted with aqueous methanol from 18 different thyme (Thymus L.) specimens and species. This evaluation is made with use of the same free radical source (DPPH• radical), three different free radical scavenging models (gallic acid, ascorbic acid, and Trolox), and three different measuring techniques (the dot blot test, UV-Vis spectrophotometry, and electron paramagnetic resonance spectroscopy, EPR). A comparison of the equivalence of these three different measuring techniques (performed with use of hierarchical clustering with Euclidean distance as a similarity measure and Wards linkage) is particularly important in view of the fact that different laboratories use different antioxidant activity measuring techniques, which makes any interlaboratory comparison hardly possible. The results obtained confirm a semiquantitative equivalence among the three compared methodologies, and a proposal is made of a simple and cost-effective dot blot test that uses the DPPH• radical and provides differentiation of antioxidant activity of herbal matter comparable with the results of the UV-Vis spectrophotometry and EPR.

Comparison of humification processes occurring during sewage purification in treatment plants with different technological processes.

The course of the humification process of sewage sludge collected from three biologic-mechanical treatment plants with different treatment technologies was studied. The maturity of sewage sludge and its usefulness for agricultural purposes was also discussed. The physical-chemical properties of humic acids extracted from sewage sludge received from comparable stages of sludge purification were described. Changes of the sludge properties during sewage purification and the progress of the humification process were investigated with EPR, IR and UV/VIS spectroscopic methods. The content of the elements and the carboxylic groups in humic acids extracted from each stage of the sewage treatment were also determined. It was found that the humification processes take place in all three treatment plants but with different intensities resulting from the differences in the individual cleaning processes in these plants. The most intensive changes of physical-chemical parameters in the extracted humic acids were observed in the anaerobic digester where mesophilic fermentation occurs. The sludge oxygenation processes also significantly affect the course of the humification process during sewage treatment.

A comparison of antioxidative capacities of fruit juices, drinks and nectars, as determined by EPR and UV-vis spectroscopies.

The differences in the Trolox Equivalent Antioxidant Capacity (TEAC) values at the same incubation time obtained by two different techniques: electron paramagnetic resonance (EPR) spectroscopy and ultraviolet visible (UV-vis) spectroscopy, which use the same antioxidant-free radical reaction mechanism, were determined for fruit juices, nectars and drinks. For this study, the stable free radical 1,1-Diphenyl-2-picryl-hydrazyl (DPPH(•)) was used. The antioxidant capacity was presented in Trolox Equivalents, e.g., μM trolox per 100 ml of sample. All of the studied fruit juices, drinks and nectars showed antioxidative properties. Dependencies between TEAC values and the percent fruit content and sample color were observed for the studied beverages. It was found that EPR spectroscopy is the more adequate method for determining TEAC values for these kinds of samples.

Thermal stability of haemoglobin solutions under dc and ac magnetic field and UV and IR radiation

The influence of electromagnetic field in wide range of frequency (extra low frequency magnetic field, monochromatic laser light 840 nm, ultraviolet light) and static magnetic field on the stability of aqueous and NaCl 0.9% haemoglobin solutions was studied using differential scanning calorimetry (DSC) method. Three steps of denaturation process can be derived from experimental data. The shifts of transition temperatures are more marked for NaCl 0.9% than for aqueous solutions. The results suggest that static magnetic field is effective in protecting protein, while ultraviolet radiation destabilises haemoglobin. The laser light and alternating magnetic field have a little effect on the thermal stability of haemoglobin.

Antioxidant Capacity: Experimental Determination by EPR Spectroscopy and Mathematical Modeling

A new method of determining antioxidant capacity based on a mathematical model is presented in this paper. The model was fitted to 1000 data points of electron paramagnetic resonance (EPR) spectroscopy measurements of various food product samples such as tea, wine, juice, and herbs with Trolox equivalent antioxidant capacity (TEAC) values from 20 to 2000 μmol TE/100 mL. The proposed mathematical equation allows for a determination of TEAC of food products based on a single EPR spectroscopy measurement. The model was tested on the basis of 80 EPR spectroscopy measurements of herbs, tea, coffee, and juice samples. The proposed model works for both strong and weak antioxidants (TEAC values from 21 to 2347 μmol TE/100 mL). The determination coefficient between TEAC values obtained experimentally and TEAC values calculated with proposed mathematical equation was found to be R(2) = 0.98. Therefore, the proposed new method of TEAC determination based on a mathematical model is a good alternative to the standard EPR method due to its being fast, accurate, inexpensive, and simple to perform.

Investigation of Antioxidant Activity of Pomegranate Juices by Means of Electron Paramagnetic Resonance and UV-Vis Spectroscopy.

Pomegranate fruit (Punica granatum L.) is a source of numerous phenolic compounds, and it contains flavonoids such as anthocyanins, anthocyanidins, cyanidins, catechins and other complexes of flavonoids, ellagitannins, and hydrolyzed tannins. Pomegranate juice shows antioxidant, antiproliferative, and anti-atherosclerotic properties. The antioxidant capacity (TEAC) of the pomegranate juices was measured using electron paramagnetic resonance (EPR) spectroscopy and 1,1-diphenyl-2-picrylhydrazyl (DPPH•) as a source of free radicals, and the total phenolic (TP) content was measured using UV-Vis spectroscopy. All the examined pomegranate juices exhibited relatively high antioxidant properties. The TEAC values determined by means of EPR spectroscopy using Trolox (TE) as a free radical scavenger were in the range of 463.12 to 1911.91 μmol TE/100 mL juice. The TP content measured by the Folin-Ciocalteu method, using gallic acid (GA) as a free radical scavenger, widely varied in the investigated pomegranate juice samples and ranged from 1673.62 to 5263.87 mg GA/1 L juice. The strongest antioxidant properties were observed with the fresh pomegranate juices obtained from the fruits originating from Israel, Lebanon, and Azerbaijan. Correlation analysis of numerical data obtained by means of EPR spectroscopy (TEAC) and UV-Vis spectroscopy (TP) gave correlation coefficient (r)=0.90 and determination coefficient (r2)=0.81 (P<0.05).