Streptomycin (STR) is an aminoglycoside antibiotic with a broad-spectrum of activity and ototoxic potential. The mechanism of STR-induced inner ear damage has not been fully elucidated. It was previously found that STR binds to melanin, which may result in the accumulation of the drug in melanin-containing tissues. Melanin pigment is present in various parts of the inner ear, including the cochlea and vestibular organ. The present study aimed to assess if streptomycin generates oxidative stress and affects melanogenesis in normal human melanocytes. Moreover the variation of free radical concentration in STR-treated melanocytes was examined by electron paramagnetic resonance spectroscopy (EPR). We found that STR decreases cell metabolic activity and reduces melanin content. The observed changes in the activity of antioxidant enzymes activity in HEMn-DPs treated with streptomycin may suggest that the drug affects redox homeostasis in melanocytes. In this work EPR study expanded knowledge about free radicals in interactions of STR and melanin in melanocytes. The results may help elucidate the mechanisms of STR toxicity on pigment cells, including melanin-producing cells in the inner ear. This is important because understanding the mechanism of STR-induced ototoxicity would be helpful in developing new therapeutic strategies to protect patients' hearing.
The antioxidant activity and the association of genistein with carcinogenesis are widely documented. Few studies directly measure the number of free radicals generated in cells, either during the action of factors stimulating their formation, e.g., ultraviolet (UV), or after exposure to antioxidants. The most suitable method for analysing free radicals is electron paramagnetic resonance (EPR) spectroscopy. The EPR method detects a paramagnetic centre with a single electron. Antioxidants neutralize free radicals, therefore, EPR analysis of antioxidant efficacy is as valuable and important as studying the paramagnetic centres of radicals. The aim of the study was to determine the influence of genistein on free radicals basal level and after UV exposure in breast cancer cell lines MCF7, T47D and MDA-MB-231 cell lines. The impact of genistein on cell viability was investigated at concentrations of 0.37 µM, 3.7 µM, 37 µM and 370 µM. Genistein at a concentration of 370 µM revealed a cytotoxic effect on the cells of all three tested breast cancer lines. Genistein at a concentration of 0.37 µM showed no significant effect on the cell viability of all tested breast cancer lines. Therefore, cell proliferation and antioxidant properties were examined using genistein at a concentration of 0.37 µM and 37 µM. X-band (9.3 GHz) EPR spectra of three different types of breast cancer cells (ER-positive, PR-positive and HER-2 negative: MCF7 and T47D and triple-negative MDA-MB-231) were compared. UV irradiation was used as a factor to generate free radicals in cells. The effect of free radical interactions with the antioxidant genistein was tested for non-UV-irradiated (corresponding to the basal level of free radicals in cells) and UV-irradiated cells. The levels of free radicals in the non-irradiated cells studied increased in the following order in breast cancer cells: T47D < MDA-MB-231 < MCF7 and UV-irradiated breast cancer cells: MDA-MB-231 < MCF7 < T47D. UV-irradiation altered free radical levels in all control and genistein-cultured cells tested. UV irradiation caused a slight decrease in the amount of free radicals in MCF7 cells. A strong decrease in the amount of free radicals was observed in UV-irradiated MDA-MB-231 breast cancer cells. The amount of free radicals in T47D cancer cells increased after UV irradiation. Genistein decreased the amount of free radicals in non-irradiated and UV-irradiated MCF7 cells, and only a weak effect of genistein concentrations was reported. Genistein greatly decreased the amount of free radicals in UV-irradiated T47D cancer cells cultured with genistein at a concentration of 3.7 µM. The effect of genistein was negligible in the other samples. Genistein at a concentration of 3.7 µM decreased the amount of free radicals in non-irradiated MDA-MB-231 cancer cells, but genistein at a concentration of 37 µM did not change the amount of free radicals in these cells. An increase in the amount of free radicals in UV-irradiated MDA-MB-231 cancer cells was observed with increasing genistein concentration. The antioxidant efficacy of genistein as a potential plant-derived agent supporting the treatment of various cancers may be determined by differences in signalling pathways that are characteristic of breast cancer cell line subtypes and differences in activation of oxidative stress response pathways.
Free radical formation in two diuretics: indapamide and torasemide was examined during UV irradiation and storage at higher temperatures using X-band (9.3â¯GHz) electron paramagnetic resonance spectroscopy (EPR). The aim of this study was to investigate the possibility of storing indapamide and torasemide under UV irradiation and at higher temperatures, which may occur during exposure to light. The diuretic samples were exposed to UVA irradiation for 15, 30 and 45â¯minutes, and stored at temperatures of 40 °C and 50 °C by 30â¯minutes. The EPR spectra were analyzed to determine the amplitudes (A), linewidths (ΔBpp), and integral intensities (I) and g factors. The concentrations of free radical (N) in the diuretic samples were also determined. The influence of microwave power on amplitudes, linewidths and the asymmetry parameter were evaluated. The result showed that the tested indapamide and torasemide samples exhibited high free radical concentrations in the range of 1018-1019 spin/g after UV irradiation and heat treatment. Therefore, due to the significant free radical formation indapamide and torasemide should not be stored under UV light and at temperatures of 40 °C and 50 °C. The complex character of free radical systems in the diuretic samples was proved as evidenced by the changes of the asymmetry parameters of the EPR lines with increasing microwave power. Fast spin-lattice relaxation processes were observed in all tested diuretic samples, regardless of the storage conditions. Electron paramagnetic resonance spectroscopy is proposed as a useful method in pharmacy to determine the appropriate storage conditions for diuretics.
Hot Temperature , Indapamide , Torsemide , Temperature , Electron Spin Resonance Spectroscopy/methods , Ultraviolet Rays , Free Radicals/chemistry , Diuretics
An X-band (9.3 GHz) electron paramagnetic resonance (EPR) spectroscopy was used to examine the free radical scavenging activity of the following infusions, which were nonirradiated and exposed to UVA: root of Asparagus racemosus and herbs of Mitchella repens, Cnicus benedictus L., Galega officinalis L., and Eupatorium cannabinum L. The plant materials for obstetrics applications were chosen for analysis. The aims of these studies were to compare the free radical scavenging ability of the tested infusions and to determine the influence of UVA irradiation of the plant materials on interactions of these infusions with free radicals. Both the magnitude and kinetics of the interactions of the infusions with the model DPPH free radicals were examined. The ability to quench the free radicals for the examined plant infusions increases in the following order: Asparagus racemosus (root) < Mitchella repens (herb) < Cnicus benedictus L. (herb) < Galega officinalis L. (herb) < Eupatorium cannabinum L. (herb). The analyzed infusions differ in the kinetics of the interactions with free radicals. The fastest interactions with free radicals characterize the infusions of Galega officinalis L. herb and Eupatorium cannabinum L. herb. The infusion of Mitchella repens herb interacts with free radicals in the slowest way. UVA radiation reduces the antioxidant interactions of all tested infusions, especially the infusion of Eupatorium cannabinum L. herb, which should be protected against UVA radiation during storage. The weakest decrease of free radical scavenging activity was observed for the infusion of the root of Asparagus racemosus exposed to UVA radiation. UVA radiation affected the speed of the free radical interactions of the infusions, depending on the type of plant materials. EPR spectroscopy is useful to examine the free radical scavenging activity of plant infusions, which is helpful to find effective antioxidants for applications in obstetrics and their optimal storage conditions.
The evolution of the paramagnetic center system in blood during the healing of skin burn wounds dressed with a biodegradable apitherapeutic nanofiber dressing was examined. The aim of this study was to determine the changes in paramagnetic centers in blood during the influence of apitherapeutic nanofiber dressings on the healing process. The blood samples were tested before burn infliction (day 0) and, respectively, on the 10th and 21st days of the experiment. Paramagnetic centers in the blood of the pig used as the model animal were examined with an X-band (9.3 GHz) electron paramagnetic resonance spectroscopy. The EPR spectra were measured with Bruker spectrometer at 230 K with a modulation frequency of 100 kHz. The EPR lines of the high spin Fe3+ in methemoglobin, high spin Fe3+ in transferrin, Cu2+ in ceruloplasmin, and free radicals were observed in the multicomponent spectra of blood. For the application of the apitherapeutic nanofiber dressing, the amplitudes of the EPR signals of Fe3+ in methemoglobin were similar up to 10 days. For the experiment with the apitherapeutic formulation, the heights of EPR signals of Fe3+ in transferrin were lower after 10 days and 21 days of therapy, compared to day 0. For the application of the apitherapeutic formulation the signals of Cu2+ in ceruloplasmin and free radicals, strongly decreased after 10 days of therapy, and after 21 days it increased to the initial values characteristic for day 0. The apitherapeutic formulation caused that after 21 days the EPR spectrum of Cu2+ in ceruloplasmin and free radicals was considerably high. The apitherapeutic formulation interaction after 10 days and after 21 days of therapy resulted in the low EPR lines of Fe3+ in methemoglobin. EPR spectra of blood may be useful for presentation of the changes in its paramagnetic centers during the healing process of the burn wounds.
Bandages/standards , Burns/drug therapy , Nanofibers/therapeutic use , Propolis/therapeutic use , Animals , Humans , Propolis/pharmacology , Swine
Malignant melanoma is the most aggressive and lethal form of skin cancer. Therefore the search for new methods of the treatment for melanoma is needed. In this work simvastatin was tested as the substance of potential anticancer activity. The aim of this study was to determine the effect of simvastatin in different concentration on free radicals in A-375 human melanoma malignum cells. Free radicals and growth of A-375 melanoma cells cultured without and with simvastatin were examined. Free radicals were tested by the use of electron paramagnetic resonance (EPR) spectroscopy. o-Semiquinone free radicals existed in A-375 cells. Free radicals of the control A-375 cells and the cells cultured with simvastatin were homogeneously broadened. The growth of A-375 cells did not change for concentrations of simvastatin 0.1⯵M and 0.3⯵M, and it slightly decreased for concentration of this substance equal 1⯵M, so these concentrations of simvastatin were not preferred for therapy. For the concentrations of simvastatin higher than 1⯵M the growth of A-375 cells was considerably weakened. Simvastatin in the concentrations of 0.1⯵M and 1⯵M decreased free radical concentrations in A-375 cells. The concentrations of simvastatin of 3⯵M and 5⯵M accompanied by the strong cell damage and strong formation of free radicals in them were recommended for anticancer therapy of A-375 melanoma cells.
Antineoplastic Agents/pharmacology , Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology , Melanoma/drug therapy , Simvastatin/pharmacology , Skin Neoplasms/drug therapy , Cell Line, Tumor , Cell Proliferation/drug effects , Electron Spin Resonance Spectroscopy , Free Radicals/metabolism , Humans , Melanoma/metabolism , Skin Neoplasms/metabolism
The multicomponent electron paramagnetic resonance spectra of the blood during healing of skin burned wounds treated with a new generation biodegradable dressings containing poly(lactide-co-glycolide) were analysed. The evolution of different types of paramagnetic centers in the blood with time of healing was determined. The EPR spectra of the blood samples at 230 K temperature were measured at 1, 10, and 21 days after burning of the pig skin. The EPR lines of the following paramagnetic centers: the high-spin Fe3+ in methemoglobin (line I), high-spin Fe3+ in transferrin (line II), and Cu2+ in ceruloplasmin and free radicals (line III) were observed in the X-band (9.3 GHz) spectra of the blood. The multicomponent structure of the EPR spectra of the tested blood samples depended on the time of the healing of the burned wounds. The amount of the high-spin Fe3+ in methemoglobin (line I) in the blood decreased after 21 days of the healing of the burned wounds. The amount of the high-spin Fe3+ in transferrin (line II) slightly increased after 21 days of therapy with the basis. The amount of Cu2+ in ceruloplasmin and free radicals (line III) in the blood was very high after 10 days of therapy. At the first day of the healing of the burned wounds, the highest amount of the high-spin Fe3+ in methemoglobin (line I), the relatively lower amounts of the high-spin Fe3+ in transferrin (line II), and Cu2+ in ceruloplasmin and free radicals (line III) existed in the blood. In the medium phase (after 10 days) of the healing of the burned wounds, the extremely higher amounts of Cu2+ in ceruloplasmin and free radicals (line III) appeared in the blood. In the last phase (after 21 days), only the low differences between the amounts of the high-spin Fe3+ in methemoglobin (line I), the high-spin Fe3+ in transferrin (line II), and Cu2+ in ceruloplasmin and free radicals (line III) were observed. The present study may serve as a starting point for the development of a new technique for monitoring molecular complexes containing iron Fe3+ (methemoglobin, transferrin) or copper Cu2+ ions (ceruloplasmin) and free radicals in the blood during wound healing.
Burns/therapy , Electron Spin Resonance Spectroscopy/methods , Animals , Swine
Ototoxicity is well-documented but not fully understood undesirable side effect of aminoglycoside antibiotic, kanamycin. Kanamycin is capable of binding to melanin biopolymers-natural pigments of the skin, hair, and eyes. Melanin-producing cells, melanocytes, are also present in the inner ear and are known to be necessary for normal hearing. It was considered that melanin content in the inner ear may influence aminoglycoside-induced ototoxic effect. The impact of kanamycin on melanocytes homeostasis may thus play role in the antibiotic-induced ototoxic effect. Previously, we demonstrated that kanamycin disturbs homeostasis in light-pigmented melanocytes. To investigate if/how melanization contributes to this phenomenon, the study using in vitro model of dark-pigmented melanocytes is required. Spectrophotometric measurements and electron paramagnetic resonance (EPR) spectroscopy analysis were performed. Kanamycin induced a concentration-dependent loss in HEMn-DP melanocytes viability. The value of IC 50 was estimated to be 5.0 mM. Modulation of the activity of analyzed antioxidant enzymes and increased production of free radicals as well as the decrease of the melanin content were observed. Our results confirmed that kanamycin generates oxidative stress in melanocytes. The increased level of free radicals caused by kanamycin may be responsible for the imbalance of antioxidant defense and the reduction of melanin content in melanocytes. The role of melanin in the mechanism of kanamycin-induced hearing impairment was discussed and the obtained results were compared with the previously demonstrated data concerning light-pigmented melanocytes.
The influence of concentration of simvastatin (SIM) on free radicals in A-2058 human melanoma malignum cells was studied. The proliferation assay for melanoma A-2058 cells with SIM in concentration range from 0.1 to 20 µM was performed. SIM in the concentrations of 0.1, 0.3, and 1 µM only slightly changed the growth of A-2058 cells, but the growth of the cells considerably decreased for higher concentrations of SIM. Free radicals in the cells were examined by an X-band (9.3 GHz) electron paramagnetic resonance (EPR) spectroscopy. o-Semiquinone free radicals with g-factors in the range of 2.0060 to 2.0065 were found in A-2058 cells. The asymmetric broad EPR spectra with linewidths (ΔBpp ) from 0.87 to 1.25 mT were measured. The fast spin-lattice relaxation processes characterized all the tested cells. The free radical concentrations in the all A-2058 cells cultured with SIM were lower than in the control cells. The quenching of free radicals in A-2058 cells depended on concentration of SIM. This effect was the weakest for concentration of SIM of 3 µM. The strongest decrease of free radical concentration caused SIM in concentration of 1 µM.
Free radicals formed during thermal sterilization of the Ungentum ophthalmicum were examined by an X-band EPR spectroscopy. The influence of storage time (15 min; 1, 2 and 3 days after heating) on free radical properties and concentrations in this sample was determined. Thermal sterilization was done according to the pharmaceutical norms. The first-derivative EPR spectra with g-values about 2 were measured with magnetic modulation of 100 kHz in the range of microwave power 2.2-70 mW. The changes of amplitudes (A) and linewidths (ΔBpp) with microwave powers were evaluated. Free radicals in concentration â¼1017 spin/g were formed during heating of the tested Ungentum. Free radical concentration decreased with increase in storage time, and reached values â¼1017 spin/g after 3 days from sterilization. The tested U. ophthalmicum should not be sterilized at a temperature of 160 °C because of the free radicals formation, or it should be used 3 days after heating, when free radicals were considerably quenched. Free radical properties remain unchanged during storage of the Ungentum. The EPR lines of the U. ophthalmicum were homogeneously broadened and their linewidths (ΔBpp) increased with increase in microwave power. EPR spectroscopy is useful to examine free radicals to optimize sterilization process and storage conditions of ophthalmologic samples.
Electron Spin Resonance Spectroscopy/methods , Free Radicals/analysis , Hot Temperature , Ophthalmic Solutions/standards , Sterilization/standards , Administration, Ophthalmic , Drug Storage/methods , Drug Storage/standards , Sterilization/methods
Free radicals in thermally treated chloramphenicol were examined by electron paramagnetic resonance (EPR) spectroscopy. The parameters and shape of EPR spectra were analysed and free radical concentrations were obtained in the tested drug samples. Chloramphenicol was thermally sterilized at pharmacopeia conditions: 100 °C (120 min). Sterilization was also carried out at different conditions, 110 °C (60 min) and 120 °C (30 min), for comparison with pharmacopeia settings. Microbiological analysis was performed on the samples to confirm sterility. The aim of this work was to determine the concentration of free radicals in chloramphenicol following thermal sterilization at pharmacopeia conditions and compare this with other sets of conditions [110 °C (60 min) and 120 °C (30 min)]. The best conditions of thermal sterilization are determined as those that kill microorganisms and produce the lowest amounts of free radicals in this drug. It was concluded that the optimal temperatures and times for the thermal sterilization of chloramphenicol are 100 °C and 120 min and 110 °C and 60 min. A temperature of 120 °C coupled with a heating time of 30 min was rejected for thermal sterilization because of the high amount of free radicals produced by the drug samples.
Anti-Bacterial Agents/chemistry , Chloramphenicol/chemistry , Free Radicals/chemistry , Hot Temperature , Microwaves , Sterilization/methods , Anti-Bacterial Agents/metabolism , Chloramphenicol/metabolism , Electron Spin Resonance Spectroscopy/methods , Free Radicals/metabolism , Spectrophotometry, Ultraviolet/methods
Normal and keloid fibroblasts were examined using X-band (9.3 GHz) electron paramagnetic resonance spectroscopy. The effect of genistein on the concentration of free radicals in both normal dermal and keloid fibroblasts after ultraviolet irradiation was investigated. The highest concentration of free radicals was seen in keloid fibroblasts, with normal fibroblasts containing a lower concentration. The concentration of free radicals in both normal and keloid fibroblasts was altered in a concentration-dependent manner by the presence of genistein. The change in intra-cellular free radical concentration after the ultraviolet irradiation of both normal and keloid fibroblasts is also discussed. The antioxidant properties of genistein, using its 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging activity as a model, were tested, and the effect of ultraviolet irradiation on its interaction with free radicals was examined. The electron paramagnetic resonance spectra of DPPH showed quenching by genistein. The interaction of genistein with DPPH free radicals in the absence of ultraviolet irradiation was shown to be slow, but this interaction was much faster under ultraviolet irradiation. Ultraviolet irradiation enhanced the free radical-scavenging activity of genistein.
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.
Burns/pathology , Electron Spin Resonance Spectroscopy/methods , Free Radicals/analysis , Numerical Analysis, Computer-Assisted , Wounds and Injuries/pathology , Animals , Microwaves , Normal Distribution , Sus scrofa
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.
Free Radical Scavengers/chemistry , Ophthalmic Solutions/radiation effects , Propolis/radiation effects , Ultraviolet Rays/adverse effects , Biphenyl Compounds/antagonists & inhibitors , Electron Spin Resonance Spectroscopy , Hot Temperature , Ophthalmic Solutions/chemistry , Oral Sprays , Oxidation-Reduction/radiation effects , Picrates/antagonists & inhibitors , Propolis/chemistry
Free radicals thermally generated in the ointments containing propolis were studied by electron paramagnetic resonance (EPR) spectroscopy. The influence of temperature on the free radical concentration in the propolis ointments was examined. Two ointment samples with different contents of propolis (5 and 7%, resp.) heated at temperatures of 30°C, 40°C, 50°C, and 60°C, for 30 min., were tested. Homogeneously broadened EPR lines and fast spin-lattice interactions characterized all the tested samples. Free radicals concentrations in the propolis samples ranged from 10(18) to 10(20) spin/g and were found to grow in both propolis-containing ointments along with the increasing heating temperature. Free radical concentrations in the ointments containing 5% and 7% of propolis, respectively, heated at temperatures of 30°C, 40°C, and 50°C were only slightly different. Thermal treatment at the temperature of 60°C resulted in a considerably higher free radical formation in the sample containing 7% of propolis when related to the sample with 5% of that compound. The EPR examination indicated that the propolis ointments should not be stored at temperatures of 40°C, 50°C, and 60°C. Low free radical formation at the lowest tested temperatures pointed out that both examined propolis ointments may be safely stored up to the temperature of 30°C.
Complex free radical system in thermally sterilized acidum boricum (AB) was studied. Acidum boricum was sterilized at temperatures and times given by pharmaceutical norms: 160 degrees C and 120 min, 170 degrees C and 60 min and 180 degrees C and 30 min. The advanced spectroscopic tests were performed. The EPR spectra of free radicals were measured as the first derivatives with microwaves of 9.3 GHz frequency and magnetic modulation of 100 kHz. The Polish X-band electron paramagnetic resonance spectrometer of Radiopan (Poznan) was used. EPR lines were not observed for the nonheated AB. The broad EPR asymmetric lines were obtained for all the heated AB samples. The influence of microwave power in the range of 2.2-70 mW on the shape of EPR spectra of the heated drug samples was tested. The following asymmetry parameters: A1/A2, A1-A2, B1/B2, and B1-B2, were analyzed. The changes of these parameters with microwave power were observed. The strong dependence of shape and its parameters on microwave power proved the complex character of free radical system in thermally sterilized AB. Changes of microwave power during the detection of EPR spectra indicated complex character of free radicals in AB sterilized in hot air under all the tested conditions. Thermolysis, interactions between free radicals and interactions of free radicals with oxygen may be responsible for the complex free radicals system in thermally treated AB. Usefulness of continuous microwave saturation of EPR lines and shape analysis to examine free radicals in thermally sterilized drugs was confirmed.
Anti-Infective Agents/radiation effects , Boric Acids/radiation effects , Electron Spin Resonance Spectroscopy , Free Radicals/radiation effects , Hot Temperature , Sterilization/methods , Technology, Pharmaceutical/methods , Anti-Infective Agents/chemistry , Boric Acids/chemistry , Chemistry, Pharmaceutical , Drug Stability , Free Radicals/chemistry , Microwaves , Spectrophotometry, Ultraviolet , Time Factors
Rosuvastatin was thermally sterilized at temperatures 160°C during 120 min, 170°C during 60 min and 180°C during 30 min, according to Polish pharmacopoeia norms. Free radicals formed in the heated rosuvastatin samples were examined by electron paramagnetic resonance (EPR) spectroscopy. The best conditions of thermal sterilization of rosuvastatin were searched. EPR spectra were measured for the analyzed drug independent on heating temperature. Decrease of free radical concentrations in rosuvastatin with increasing of heating temperature was stated. The optimal conditions for thermal sterilization of rosuvastatin were temperature 180°C and heating time of 30 min and resulted in the lowest free radical concentration. The changes of amplitudes (A) and linewidths (∇Bpp) of EPR spectra of the heated rosuvastatin were characteristic for homogeneous broadening EPR lines. Slow spin-lattice relaxation processes existed in the all rosuvastatin samples. The confirmation of usefulness of electron paramagnetic resonance spectroscopy in pharmacy was presented.
Electron Spin Resonance Spectroscopy/methods , Free Radicals/chemistry , Rosuvastatin Calcium/chemistry , Sterilization/methods , Hot Temperature , Pharmaceutical Services , Pharmacopoeias as Topic , Temperature , Time Factors
Free radicals formation in the acidum boricum (AB) during thermal sterilization process was examined by an X-band (9.3 GHz) electron paramagnetic resonance (EPR) spectroscopy. Acidum boricum was sterilized according to the pharmacopea norms at temperatures 160°C (120 min), 170°C (60 min), and 180°C (30 min). Free radicals (~10(17) spin/g) were thermally formed in these drug. The free radicals system revealed complex character, and the asymmetrical EPR spectra were measured. Mainly oxygen free radicals exist in the tested heated AB. Slower spin-lattice relaxation processes exist in AB sterilized at 160, 170 and 180°C. AB may be sterilized at temperatures 160, 170 and 180°C. For AB thermal sterilization at temperature 170°C is recommended. Free radicals concentrations changes during storage of the examined AB, and probably interactions with oxygen molecules may be responsible for this effect.
Boron Compounds/chemistry , Sterilization , Electron Spin Resonance Spectroscopy , Free Radicals , Hot Temperature
Electron paramagnetic resonance (EPR) spectroscopy was used for examination of free radicals in thermally treated vaselinum album (VA). Thermal treatment in hot air as sterilization process was tested. Conditions of thermal sterilization were chosen according to the pharmaceutical norms. Vaselinum album was heated at the following conditions (T--temperature, t--time): T = 160°C and t = 120 min, T = 170°C and t = 60 min and T = 180°C and t = 30 min. The aim of this work was to determine concentration and free radical properties of thermally sterilized VA. EPR analysis for VA was done 15 min after sterilization. EPR measurements were done at room temperature. EPR spectra were recorded in the range of microwave power of 2.2-70 mW. g-Factor, amplitudes (A) and line width (ΔBpp) of the spectra were determined. The shape of the EPR spectra was analyzed. Free radical concentration (N) in the heated samples was determined. EPR spectra were not obtained for the non heated VA. EPR spectra were detected for all thermally sterilized samples. The spectra revealed complex character, their asymmetry depends on microwave power. The lowest free radicals concentration was found for the VA sterilized at 180°C during 30 min. EPR spectroscopy is proposed as the method useful for optimization of sterilization process of drugs.
Electron Spin Resonance Spectroscopy , Petrolatum/chemistry , Free Radicals , Hot Temperature , Ointments , Sterilization
Electron paramagnetic resonance (EPR) spectroscopy may be applied to examine interactions of melanin with metal ions and drugs. In this work EPR method was used to examination of changes in free radical system of DOPA-melanin--the model eumelanin after complexing with diamagnetic cadmium(II) ions. Cadmium(II) may affect free radicals in melanin and drugs binding by this polymer, so the knowledge of modification of properties and free radical concentration in melanin is important to pharmacy. The effect of cadmium(II) in different concentrations on free radicals in DOPA-melanin was determined. EPR spectra of DOPA-melanin, and DOPA-melanin complexes with cadmium(II) were measured by an X-band (9.3 GHz) EPR spectrometer produced by Radiopan (Poznan, Poland) and the Rapid Scan Unit from Jagmar (Krak6w, Poland). The DOPA (3,4-dihydroxyphenylalanine) to metal ions molar ratios in the reaction mixtures were 2:1, 1:1, and 1: 2. High concentrations of o-semiquinone (g ~2.0040) free radicals (~10(21)-10(22) spin/g) characterize DOPA-melanin and its complexes with cadmium(II). Formation of melanin complexes with cadmium(II) increase free radical concentration in DOPA-melanin. The highest free radical concentration was obtained for DOPA-melanin-cadmium(II) (1:1) complexes. Broad EPR lines with linewidths: 0.37-0.73 mT, were measured. Linewidths increase after binding of cadmium(II) to melanin. Changes of integral intensities and linewidths with increasing microwave power indicate the homogeneous broadening of EPR lines, independently on the metal ion concentration. Slow spin-lattice relaxation processes existed in all the tested samples, their EPR lines saturated at low microwave powers. Cadmium(II) causes fastening of spin-lattice relaxation processes in DOPA-melanin. The EPR results bring to light the effect of cadmium(II) on free radicals in melanin, and probably as the consequence on drug binding to eumelanin.
Cadmium/pharmacology , Dihydroxyphenylalanine/analogs & derivatives , Electron Spin Resonance Spectroscopy/methods , Dihydroxyphenylalanine/chemistry , Free Radicals
