Paweł Ramos

Positive Effect of Propolis on Free Radicals in Burn Wounds

Concentration and properties of free radicals in the burn wounds treated with propolis were examined by the use of electron paramagnetic resonance spectroscopy. Magnetic spin-spin interactions and complex free radicals structures in wound beds were studied. The results were compared to those obtained for silver sulphadiazine used as a standard pharmaceutical agent. The changes of free radicals in the matrix of injury with time of exposition on these substances were tested. The aim of this study was to check the hypothesis about the best influence of propolis on the burn wounds healing. It was confirmed that a relatively lower concentration of free radicals exists in the burn wounds treated with propolis. The homogeneously broadened spectra and a complex free radical system characterize the tested tissue samples. The fastening of spin-lattice relaxation processes in the matrix of injury after treatment with propolis and silver sulphadiazine was observed. Practical usefulness of electron paramagnetic resonance spectroscopy in alternative medicine was proved.

Antioxidative properties of bee pollen extracts examined by EPR spectroscopy

Antioxidative Properties of Bee Pollen Extracts Examined by EPR Spectroscopy Bee pollen is a valuable and highly recognized source of exogenous antioxidants. The aim of these studies was to determine the antioxidant capacity of three types of bee pollen extracts: ethanol extracts of bee pollen, pepsin extracts of bee pollen and ethanol extracts of pepsin-digested bee pollen. Their antioxidant properties were determined with the use of electron paramagnetic resonance (EPR) and their ability to quench DPPH free radicals was estimated. The EPR results showed that ethanol extracts of pepsin-digested bee pollen (EEPP) had the highest antioxidative effect and the highest free radical DPPH scavenging potential. The pepsin extracts of bee pollen (PEP) had the weakest antioxidant capacity. The ability to quench DPPH free radicals was also the weakest one for this extract. An average antioxidative effect was recorded for ethanol extracts of bee pollen (EEP). Właściwości Antyoksydacyjne Ekstraktów z Pyłku Pszczelego Badane Metodą Spektroskopii EPR W ostatnim czasie dużo uwagi poświęca się substancjom o działaniu antyoksydacyjnym pochodzenia naturalnego. Cennym i uznanym źródłem egzogennych antyoksydantów jest pszczeli pyłek kwiatowy. W związku z powyższym, przedmiotem badań było oznaczenie aktywności antyoksydacyjnej trzech rodzajów ekstraktów z pyłku pszczelego: ekstraktu etanolowego pyłku pszczelego, ekstraktu pepsynowego pyłku pszczelego oraz ekstraktu etanolowego po uprzednim trawieniu enzymatycznym pepsyną pyłku pszczelego. Właściwości antyoksydacyjne oznaczono metodą elektronowego rezonansu paramagnetycznego (EPR) na podstawie zdolności redukcji wolnego rodnika DPPH. Najwyższą aktywnością antyoksydacyjną oznaczoną metodą EPR charakteryzował się ekstrakt etanolowy po uprzednim trawieniu enzymatycznym pepsyną pyłku pszczelego (EEPP). Dla tego rodzaju ekstraktu zdolność redukcji wolnego rodnika DPPH była najsilniejsza. Z kolei ekstrakt pepsynowy pyłku pszczelego (PEP) cechował się najniższą aktywnością antyoksydacyjną. Zdolność redukcji wolnego rodnika DPPH dla tego ekstraktu była najsłabsza. Pośrednią aktywność antyoksydacyjną spośród badanych ekstraktów wykazano dla ekstraktu etanolowego pyłku pszczelego (EEP).

Application of Electron Paramagnetic Resonance Spectroscopy to Comparative Examination of Different Groups of Free Radicals in Thermal Injuries Treated with Propolis and Silver Sulphadiazine

Different groups of free radicals expressed in burn wounds treated with propolis and silver sulphadiazine were examined. The thermal effect forms major types of free radicals in a wound because of the breaking of chemical bonds. Free radicals, located in the heated skin, were tested after 21 days of treating by these two substances. The aim of this work was to find the method for determination of types and concentrations of different groups of free radicals in wound after high temperature impact during burning. The effects of the therapy by propolis and silver sulphadiazine on free radicals were studied. Since the chemical methods of free radicals studies are destructive, the usefulness of the electron paramagnetic resonance spectroscopy was tested in this work. The electron paramagnetic resonance spectra measured with the microwave power of 2.2 mW were numerically fitted by theoretical curves of Gaussian and Lorentzian shapes. The experimental electron paramagnetic resonance spectra of tissue samples are best fitted by the sum of one Gauss and two Lorentz lines. An innovatory numerical procedure of spectroscopic skin analysis was presented. It is very useful in the alternative medicine studies.

Microwave Saturation of Complex EPR Spectra and Free Radicals of Burnt Skin Treated with Apitherapeutic Agent

The effect of microwave power on the complex electron paramagnetic resonance spectra of the burn matrix after the therapy with propolis was examined. The spectra were measured with microwaves in the range of 2.2–79 mW. Three groups of free radicals were found in the damaged skin samples. Their spectral lines evolve differently with the microwave power. In order to detect these free radical groups, the lineshape of the spectra was numerically analysed. The spectra were a superposition of three component lines. The best fit was obtained for the deconvolution of the experimental spectra into one Gauss and two Lorentz lines. The microwave power changes also the lineshape of the spectra of thermally injured skin treated with the conventional agent—silver sulphadiazine. The spectral changes were different for propolis and for silver sulphadiazine. The number of individual groups of free radicals in the wound bed after implementation of these two substances is not equal. It may be explained by a higher activity of propolis than of silver sulphadiazine as therapeutic agents.

Application of EPR spectroscopy to the examination of pro-oxidant activity of coffee.

Free radicals present in coffee may be responsible for exerting toxic effects on an organism. The objectives of this work were to compare free radicals properties and concentrations in different commercially available coffees, in solid and liquid states, and to determine the effect of roasting on the formation of free radicals in coffee beans of various origins. The free radicals content of 15 commercially available coffees (solid and liquid) was compared and the impact of processing examined using electron paramagnetic resonance (EPR) spectroscopy at X-band (9.3 GHz). First derivative EPR spectra were measured at microwave power in the range of 0.7-70 mW. The following parameters were calculated for EPR spectra: amplitude (A), integral intensity (I), and line-width (ΔBpp); g-Factor was obtained from resonance condition. Our study showed that free radicals exist in green coffee beans (10(16) spin/g), roasted coffee beans (10(18) spin/g), and in commercially available coffee (10(17)-10(18) spin/g). Free radical concentrations were higher in solid ground coffee than in instant or lyophilised coffee. Continuous microwave saturation indicated homogeneous broadening of EPR lines from solid and liquid commercial coffee samples as well as green and roasted coffee beans. Slow spin-lattice relaxation processes were found to be present in all coffee samples tested, solid and liquid commercial coffees as well as green and roasted coffee beans. Higher free radicals concentrations were obtained for both the green and roasted at 240 °C coffee beans from Peru compared with those originating from Ethiopia, Brazil, India, or Colombia. Moreover, more free radicals occurred in Arabica coffee beans roasted at 240 °C than Robusta. EPR spectroscopy is a useful method of examining free radicals in different types of coffee.

Antioxidant Properties of Water Extracts of Sterilized and Unsterilized Morus alba L. Leaves

The electron paramagnetic resonance tests carried out indicated that both the technique and the conditions of sterilization of dried white mulberry leaves were chosen correctly, as no significant differences were found between the concentration and the properties of free radicals in the analyzed samples. The high temperature short time method steam sterilization process, the temperature, as well as the time of exposure to it during the preparation of water extracts resulted in no significant change in the content of polyphenol compounds in the samples analyzed. However, the total antioxidant activity increased, which suggested that despite technological processing mulberry leaves do not lose their antioxidative properties.

EPR examination of free radicals thermally formed in vaselinum flavum

Abstract The popular pharmaceutical base used in pharmacy – vaselinum flavum – was studied by an X-band (9.3 GHz) EPR spectrometer in the range of microwave power of 2.2–70 mW. The samples were sterilized in hot air oven at temperatures: 160°C (120 min), 170°C (60 min), and 180°C (30 min). The aim of this work was to determine properties and free radical concentrations in vaselinum flavum thermally sterilized at different conditions. The changes in free radical system in vaselinum flavum during storage were analyzed. Free radicals were found in all the heated samples. The lowest free radical concentration was obtained for vaselinum flavum heated at 180°C for 30 min; so these parameters are proposed for the thermal sterilization of this pharmaceutical base. Interactions with oxygen decreased free radical concentration in vaselinum flavum during storage. Strong quenching of free radicals in vaselinum flavum was observed after 2 days for the samples sterilized at temperatures 160 and 180°C. Such an effect for vaselinum flavum heated at temperature 170°C was observed later, 13 days after sterilization. Fast spin-lattice relaxation processes exist in thermally sterilized vaselinum flavum. The EPR lines of heated vaselinum flavum were homogeneously broadened. EPR spectroscopy and its use for examining the thermal sterilization process in pharmacy was confirmed.

Effect of UV Irradiation and Temperature on Free Radical Properties in Dehydrocholic and Ursodeoxycholic Acids: An EPR Study

The effect of UV irradiation and temperature on the formation and properties of free radicals in two pharmaceutical important bile acids, such as dehydrocholic (DH) and ursodeoxycholic acids (UDC), was examined. Electron paramagnetic resonance (EPR) spectroscopy was applied to determine the paramagnetic character of UV irradiated and thermally sterilized drugs. Thermal and UV irradiation sterilizations of both compounds were carried out at different conditions according to pharmaceutical norms. The performed EPR measurements of UV irradiated and thermally sterilized DH and UDC samples proved the existence of the complex free radical systems in examined bile acids. Significant influence of UV irradiation in comparison with applied thermal sterilization on free radical concentrations in DH and UDC samples was observed. The results pointed out that thermal method is most suitable for bile acid sterilization. Therefore, this kind of sterilization should be applied in practice.

Application of Numerical Analysis of the Shape of Electron Paramagnetic Resonance Spectra for Determination of the Number of Different Groups of Radicals in the Burn Wounds

Background. The evidence exists that radicals are crucial agents necessary for the wound regeneration helping to enhance the repair process. Materials and methods. The lineshape of the electron paramagnetic resonance (EPR) spectra of the burn wounds measured with the low microwave power (2.2 mW) was numerically analyzed. The experimental spectra were fitted by the sum of two and three lines. Results. The number of the lines in the EPR spectrum corresponded to the number of different groups of radicals in the natural samples after thermal treatment. The component lines were described by Gaussian and Lorentzian functions. The spectra of the burn wounds were superposition of three lines different in shape and in linewidths. The best fitting was obtained for the sum of broad Gaussian, broad Lorentzian, and narrow Lorentzian lines. Dipolar interactions between the unpaired electrons widened the broad Gaussian and broad Lorentzian lines. Radicals with the narrow Lorentzian lines existed mainly in the tested samples. Conclusions. The spectral shape analysis may be proposed as a useful method for determining the number of different groups of radicals in the burn wounds.

Free Radical Scavenging Activity of Drops and Spray Containing Propolis—An EPR Examination

The influence of heating at a temperature of 50 °C and UV-irradiation of propolis drops and spray on their free radical scavenging activity was determined. The kinetics of interactions of the propolis samples with DPPH free radicals was analyzed. Interactions of propolis drops and propolis spray with free radicals were examined by electron paramagnetic resonance spectroscopy. A spectrometer generating microwaves of 9.3 GHz frequency was used. The EPR spectra of the model DPPH free radicals were compared with the EPR spectra of DPPH in contact with the tested propolis samples. The antioxidative activity of propolis drops and propolis spray decreased after heating at the temperature of 50 °C. A UV-irradiated sample of propolis drops more weakly scavenged free radicals than an untreated sample. The antioxidative activity of propolis spray increased after UV-irradiation. The sample of propolis drops heated at the temperature of 50 °C quenched free radicals faster than the unheated sample. UV-irradiation weakly changed the kinetics of propolis drops or spray interactions with free radicals. EPR analysis indicated that propolis drops and spray should not be stored at a temperature of 50 °C. Propolis drops should not be exposed to UV-irradiation.