Abstracts:
3rd International Symposium in O3 Ozone Applications - June 27th 24th to 30th, 2000
OZ-PO-057: CHARACTERIZATION AND COMPARISON OF THREE SYSTEMS OF COCONUT OIL O3 ozonized.
Ozone Research Center.
Coconut oil is composed of a 97% triglycerides which are composed of saturated and unsaturated fatty acids which influence their physical and chemical characteristics. The reaction of ozone with unsaturated fatty acids from coconut oil generates the formation of fragments with carbonyl groups, carboxylic and peroxy, which can be related to their biological activity.
In the present study were characterized and compared statistically the three systems (coconut oil, coconut oil and coconut oil water with ethanol), for this analysis techniques were used in determining volmétricas peroxide, acidity, as well as an analytical technique for measuring viscosity. Furthermore, identified by NMR 1 H and the reaction products were tested for antimicrobial activity of the three systems studied. Ozonized coconut oil with ethanol reached higher levels of peroxides and acidity when compared with other systems.
This result suggests that there is greater decomposition of peroxides leading to increased formation of acids. The coefficientes of variation in both indices were lower than 10% showing high accuracy in the determinations. Were identified by NMR 1 H reaction products which were ozonides, hydroperoxides and aldehydes. Antimicrobial activity was evaluated in the three tested products having a greater spectrum of activity ozonized coconut oil with ethanol.
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OZ-PO-058: DETERMINATION OF O3 OLEOZON ® POLAR COMPOUNDS BY LIQUID - LIQUID EXTRACTION AND GAS CHROMATOGRAPHY.
Ozone Research CenteR
National Center for Scientific Research.
The components of the polar fraction of OLEOZON ® Were Identified and Quantified by a combination of a liquid-liquid extraction method with Gas Chromatographic (GC) and Gas Chromatographic-Mass Spectrometry Techniques (GC-MS).Saturated and unsaturated aldehydes and carboxylic acids I were the main products found. Hexanal, nonanal and 3-nonenal I were the main components of the polar fraction of OLEOZON ® .
The aldehyde concentration Higher Than Was an order the respective carboxylic acid. Good linearity WAS Proven Within The concentration range from 0.01 to 0.1 mol / L for aldehydes and from 0.001 to 0.01 mol / L for carboxylic acids, According To the values of the correlation coefficient (> 0.999). The values of recovery Obtained in the assay accuracy Were Higher Than 90% for all the aldehydes and carboxylic acids Assayed.
The variation from the repeatability study Coefficients Were lower than 2% for all the Studied compounds. The detection limit for Both kind of compounds Was In the Same order (10-6 mol / L) and lower than the orders Some concentration of the aldehydes and carboxylic acids in the real sample (10-3 -10 to 1 mol / L) .
The analytical method employed WAS linear, accurate, precise and sensitive Studied under the Conditions and allowed the identification of the compounds present in the Different polar fraction of OLEOZON ® .
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OZ-PO-059: ON THE ANTIBACTERIAL ACTIVITY OF O3 OLEOZON ® Gram-Positive Species. Collection strains and clinical isolates multiresistant.
Ozone Research Center,
National Center for Scientific Research.
Ozonized sunflower oil, OLEOZON ® has proven to be an effective antimicrobial agent against various bacterial species.The aim of this work was to study the in vitro activity of OLEOZON ® on Gram-positive species, both collection strains as multidrug-resistant crops. Concentrations were determined Minimum Inhibitory and Bactericidal (MIC and MBC).
In addition, studies of mortality simultaneous kinetic and electron microscopy. The results indicated that the OLEOZON ® is effective against all strains under study. The MICs were between 1.18 and 9.5 mg / mL, while the CMB were found in the range of 46.8 to 570 mg / mL.
The process of mortality showed a first order kinetics with respect to the bacterial concentration.The time required for the lethality of the species tested was in the range of 60 min to 4 h. Electron microscopy studies showed greater damage to the cell cytoplasm.
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OZ-PO-060: IN VITRO AND IN VIVO EFFECTS OF ORAL O3 OLEOZON ® Giardia lamblia trophozoites ON.
Institute of Basic and Preclinical Sciences,
Ozone Research Center,
National Center for Scientific Research,
Institute Tropical Medicine.
In the present paper in vitro and in vivo studies Were Carried out. Firstly, the in vitro inactivation capacity of OLEOZON ®on Giardia lamblia trophozoite cultures WAS Evaluated. Mean and maximum inactivation doses under controlled Conditions Were Determined, and the effect of OLEOZON ® on the Giardia lamblia ultrastructure by electron microscopy WAS Evaluated.
In the second part of This work, Giardia lamblia trophozoites inactivation in inoculated gerbils WAS Evaluated Treated with OLEOZON ® . The effect of the Treatment on Several biochemical parameters in blood (glutathione peroxidase, catalase, glutathione S-transferase, Reduced gluthatione, superoxide dismuthase, conjugated dienes, uric acid, reactive Substances to thiobartituric acid and glucose 6-phosphate dehydrogenase) Was Measured.
Results show That, Depending on Dose and microorganism concentration, OLEOZON ® inactivates Giardia lamblia trophozoites by Damaging the cell structure provoking the death of parasites. No significative change in biochemical parameters Observed WAS.
Under the Conditions of This experimental study, OLEOZON ® Showed to be Highly Effective in Disinfecting and inactivation of Giardia lamblia trophozoites and Its Effectiveness In This experimental model similar to the WAS Obtained by metronidazole.
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OZ-P-128: DICARBOXYLIC ACIDS AND THEIR ORIGINS IN THE ORAL ADMINISTRATION OF O3 OLEOZON®.
Ozone Research Center.
The principal metabolites found in the urinary excretion product of the oral administration of the OLEOZON® (ozonized sunflower oil) in Wistar rats are the dicarboxylic acids between 7 and 12 carbon atoms. These acids come from the oleic and linoleic acid, main components of the triglycerides of the sunflower oil, after their ozonization and metabolism.
The ozonides and substances (aldehydes and peroxides) with structures of 9 and 12 carbon atoms obtained by ozonolysis of the oil were converted in dicarboxylic acids by a metabolic process in which participate different enzymes and the well known mechanism of b -oxidation of fatty acids.
The aim of this work is to study the profile of urinary organic acids in rats orally treated with patterns of ozonized triglycerides: the triolein and the trilinolein using the gas chromatography-mass spectrometry to check the hypothesis of the origin of the dicarboxylic acids starting from these two triglycerides.
There were significant differences among the groups treated with the different ozonized patterns. The dicarboxylic acids present in increased quantities in the group treated with ozonized triolein were pimelic and azelaic acids; while in the group of ozonized trilinolein besides the previous ones the acids were: octenedioic, octanedioic, decenedioic and dodecenedioic.
These obtained results agree with those achieved in the study of urinary organic acid profile in rats orally treated with OLEOZON® , demonstrating that the origin of the dicarboxylic acids in the ingestion of this drug come from the oleic and linoleic acid, in principle.
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OZ-P-129: fungicidal activity of O3 OLEOZON ® FROM SPECIES OF THE GENUS CANDIDA .
Ozone Research Center,
Institute of Tropical Medicine,
National Center for Scientific Research,
Hospital Ginecobstétrico.
Candidiasis is a very common disease in the population. It was found that ozonized sunflower oil, OLEOZON ® , has antifungal activity on Candida albicans and Candida tropicalis .
The aim of this study was to determine the antifungal activity of OLEOZON ® on species of Candida as C. albicans , C. krusei , C. glabrata , C. tropicalis , C. parapsilosis ,C. lusitaniae and C. kefir .
We analyzed 132 strains, of which 80% were isolated from patients with acquired immunodeficiency syndrome (AIDS). For this purpose were determined Minimum Fungicide Concentration (MFC). Studies were conducted electron microscopy coupled to the kinetics of mortality.
The results showed that OLEOZON ® has fungicidal effects on yeast species analyzed in this paper. CPM were obtained ranging in the range of 2.92 to 356 mg / mL. The process of mortality showed a first order kinetic and electron microscopy reflected damage level cell cytoplasm.
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OZ-P-130: EVALUATION OF O3 OLEOZON ® TOXICITY TEST IN CLASS.
National Center for Laboratory Animal Production, Center for Toxicology and Animal Experimentation,
Ozone Research Center.
The toxicity test class is an alternative assay in vivo for classifying the substance as its range of toxicity. In this case allowed to classify the OLEOZON ® . For which 12 rats were used in the subline Cenp: SPRD randomly distributed at the rate of six animals per experimental group, three of each sex.
The animals were housed in plastic boxes of polycarbonate (Makrolon) type Tecniplast T 4 , maintained under conditions suitable for the species, with a temperature of 20 ± 4 ° C, relative humidity 70 ± 10% and photoperiod 12 h light-12 h darkness. The dosing was performed in a single dose via oral gavage at a rate of 2 000 mg / kg body weight of animals prior fasting for 12 h.
The control group was administered the vehicle (sunflower oil) and treated (OLEOZON ® ). Were recorded daily clinical signs and body weight of individual animals prior to administration, and at 7 and 14 days of the trial.
This substance caused no deaths or clinical signs, there was no difference in body weight, showed no gross lesions in cavities or organs at this dose. In sane with the Classification System of Acute Toxicity, according to the classification system, the substance is not classified, the DL50 is> 2 000 mg / kg body weight.
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OZ-P-131: O3 ozonized theobroma oil AND DEXAMETHASONE ON THE REACTION IN RATS passive cutaneous anaphylaxis.
Ozone Research Center.
It has been found that the ozone administered rectally in mice significantly decreased the anaphylactic reaction, for this reason we decided to evaluate the suppressive effect of ozonized theobroma oil and dexamethasone (Dx) in Balb / c by developing the reaction of Anaphylaxis Passive Cutaneous in rats.
The mice were divided into five groups of 10 animals each. Mice of groups (II, III, IV and V) were sensitized with a solution of 0.5 mg of ovalbumin and 200 mg of KAl (SO 4 ) 2 · 12 H 2 O / kg. The initial dose of awareness was repeated at 30 d.
The first group (negative control), the second group (positive control), which received only awareness, treatment groups were the III, IV and V (ozonized theobroma 9 mg / animal vr), dexamethasone (Dx) 0.5 mg / kg by intraperitoneal route and virgin theobroma oil (9 mg / animal vr) respectively for 20 d.
Sera from animals were obtained and stored at -70 or C. Subsequently proceeded to the anaphylactic skin test male Sprague Darley. Performed 30 min after intravenous injection of a solution of albumin with Evans blue was carried out the quantitative determination of dye contained in each of reaction sites.
The results indicated that there is a statistically significant decrease in anaphylactic reaction in the group treated with ozonized theobroma oil, compared with the obtained in the positive control group, however there are also significant differences with respect to the group treated with dexamethasone. Therefore it is suggested that the substance tested was ozonated capable of intervening in the synthesis of specific anti-ovalbumin IgE.
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OZ-P-132: HEALTH RECORD ON APPLICATION OF ozonized sunflower oil O3 "OLEOZON ® "IN THE epidermophytosis.
Ozone Research Center.
The vegetable oils have become means for the treatment with ozone. To ozonate these oils results in a series of chemical compounds which possess a germicidal character. The aim of our study was to obtain the sanitary registration of ozonized sunflower oil OLEOZON ® for the treatment of epidermophytosis.
The culmination of veterinary led several steps: 1-Chemical-Pharmaceutical Studies (assembly and development of different techniques, study of stability, etc..). 2-Toxicological (irritability dermal phototoxicity, sensitization test, acute toxicity and subchronic dermal, genotoxic evaluation). 3-Pharmacological studies (antifungal and antibacterial activity, effect on dermatomycosis of the rabbit, healing action).
4-Clinical trial.
They conducted a randomized controlled phase III, randomized, open, which compared the OLEOZON ® with nizoral, in patients with epidermophytosis. Group A (100 patients) received topical application of OLEOZON ® , 2 times a day for six weeks. Group B received treatment with Nizoral, the same number of times and for a like period of time that the above product. Efficacy was evaluated clinically (disappearance of all lesions, with or without mycological examination negativity) and microbiologically (microbiolólgico test negativity).
The overall assessment of the study, after 6 weeks of treatment, showed 75 and 81% efficiency for OLEOZON ® and Nizoral, respectively, without significant differences between groups. There were no cases of infection on added or adverse reactions.
One may note that both drugs have equal efficacy in the treatment of epidermophytosis. Registration was obtained OLEOZON ® for the treatment of epidermophytosis the May 12, 1999 with registration number 1498.
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OZ-P-133: O3 OLEOZON ® EN LA periodontitis.
Centre Estomatológico "III Congress of the CCP",
Centro de Investigaciones of Ozone.
Periodontitis is a chronic immunoinflammatory process initiated by periodontogénicos germs that are part of the subgingival microbiota. Surgical treatment alone can not resolve the cure and prevention of recurrence of the disease, requiring action to assist their agents to control bacterial plaque. We report a case which is used OLEOZON ®, as a germicide, to assess their effects on the treatment of simple moderate periodontitis and preventing their recurrence.
Was applied OLEOZON ® topically to the patient in the study, the area underwent surgery and at 7, 14 and 21 d after surgery in the adjacent periodontal tissues.
He took a control patient which underwent conventional treatment with chlorhexidine (0.2% aqueous solution). Both were applied to analyze the rate Health (IAH), clinical examination, radiographic and microbiological controls, at baseline, 3 and 6 months of treatment.
There was a better course and healing in the patient treated with OLEOZON ® . So far there has been no relapse, which appeared in the control patient at 6 months post-treatment. There were no adverse reactions.
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OZ-P-134: EFFECTIVENESS OF O3 OLEOZON ® AT-induced mucositis cytostatic CHILDREN.
Centre for Research and Biological Assessment, Institute of Food and Drugs,
National Institute of Oncology and Radiobiology.
In patients oncopediátricos due to repeated use of cytostatics, are ulcerations in the oral cavity and gastrointestinal tract (mucositis). In his treatment hibitane and nystatin are used. As a new therapeutic alternative proposes the ozonated oil.
A study of 36 children affected with cancer at the National Institute of Oncology and Radiobiology. They were divided into two experimental groups, one treated with OLEOZON ® and the other with hibitane + nystatin, evaluating different clinical indicators.
In 63% of those treated with traditional therapies ulcerations disappeared after 8 d of treatment with six daily cures, whereas OLEOZON ® 84%, with a maximum of three priests at 5 d showed a complete cure. The results indicate that the OLEOZON ® presents a superior clinical effectiveness with respect to hibitane and nystatin.
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OZ-P-135: IMPLEMENTATION OF NURSING O3 OLEOZON ® as alternative medicine. THERAPEUTIC AND ECONOMIC RESULTS.
Establishment Plant Havana Meat Company,
Ozone Research Center.
A study in 24 patients with pyoderma, epidermophytosis, furunculosis, ulcers and traumatic wounds using a traditional method (antibiotic and antifungal ointments and cures dry) and OLEOZON ® .
In the first quarter were treated 9 patients with the traditional method with a total of 87 d of work lost by medical certificates. There were 5.6% of total absenteeism by 3.7% of medical certificates, of which 3.3% corresponded to these 9 patients. This benefit is paid $ 224.00 in medical certificates.
In the second quarter were treated 15 patients with OLEOZON ® have not they need to present medical certificates and leave the job, healing all in less time compared with the group treated with tradicionles methods.
Absenteeism was 4.9% to 3.8% of which belonged to medical certificates, but none corresponded to the cases treated with OLEOZON ® . There were no recurrences in this group.
It saved $ 371.80 by way of medical. Therefore, decreased absenteeism thus obtained an increased production with consequent economic gain.
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OZ-P-136: A SOLUTION TO THE FOOT epidermophytosis.
Teaching HospitaL,
Ozone Research Center.
The dermatoficias fungal infections are sometimes resistant to treatment and enrolled in many cases with relapses, causing limitations and disability. Imidazole derivatives, broad-spectrum antifungal, expensive, therefore the search for new drugs for this disease still remains.
Thanks to the fungicidal effect of ozonized sunflower oil (OLEOZON ® ) presents its application in the treatment of epidermophytosis feet, whose goal was to generalize its use in patients with this condition treated at the Hospital "Carlos J. Finlay "in 1998-1999, from outpatient medical examinations for health screening, medical fees and entered.
504 patients were treated, curing 437 patients (86.7%). The most effective response, according to the clinical form was the scaly and macerated. There were no adverse effects.
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OZ-P-137: TREATMENT OF OIL O3 ozonized pyoderma.
Teaching Hospital,
Ozone Research Center.
The pyoderma are skin conditions caused by pyogenic bacteria corresponding to 90% to Staphylococcus aureus coagulase positive and Streptococcus beta haemolyticus . Based on the best results obtained in preliminary studies with ozonized oil compared with neomycin ointment in primary and secondary pyoderma, a decision is widely applied.
1098 patients were treated for pyoderma (1995 to 1999) with ozonized oil twice a day for 10 days. Healing was obtained in 997 patients (90.8%) without adverse effects.
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OZ-P-138: IMPLEMENTATION OF O3 OZONE IN HIGH pyoderma.
Clinic Natural and Traditional Medicine.
The ozone-oxygen mixture has demonstrated bactericidal, fungicidal and virucidal, being useful for treating diseases in which bacteria are involved. For this reason, we conducted a prospective open clinical trial in 190 patients with secondary piodermites diagnosed clinically and confirmed by the isolation of morphogenera and species of the causative agents by bacteriological examinations. These patients were selected outpatients between 1996 and 1997, randomly assigning them simple, without replacement, two treatment groups.
Group 1 was applied fomentations of potassium permanganate (to 1 x 2 000) for 15 min three times a day and alibour cream.
Group 2 was applied building ozonated water for 15 min three times a day. In both groups, treatment lasted 3 weeks, with weekly assessments for 21 d.
The results showed a better response in the group treated with ozonated water, yielding a 92.9% efficiency and a higher cure rate in the first week. There were no adverse reactions to ozonated water.
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Studies of the O3 Ozonized Vegetable Oils used in Medicine and Cosmetology.
Díaz M.
The ozonation of sunflower oil, coconut oil and theobroma oil have been studied, during several years, in the Ozone Research Centre. These oils have been converted in adequate carrier for ozone therapy, which have a great germicide power, being useful in Dermatology, Parasitology and Cosmetology.
When vegetable oils, constituted fundamentally by triglycerides, are ozonized, several substances as ozonides, aldehydes, and peroxides are formed, these could be related with their biological effect. The aim of this work is to show a general panorama of the research in Cuba on ozonized vegetable oils and its use in the fields of medicine and cosmetology.
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O3 Ozonation of Sunflower Oil: Spectroscopic Monitoring of the Degree of Unsaturation.
Soriano N.U., Migo V.P. and Matsumura M. (Japan)
Consumption of ozone by unsaturated fatty acid moieties of sunflower oil (SFO) was monitored by 1H NMR and FT-IR spectroscopy. Degradation of linoleate was found to be 1.5 and 1.8 times higher than oleate when SFO was ozonized in the absence and presence of water, respectively.
Products of ozonation in both cases include aldehyde and ozonide with 1,2,4-trioxolane ring. Hydroxyl containing compounds, which could be carboxylic acids and/or hydroperoxides, were also detected in samples ozonized in the presence of water.
The extent of ozonation had very little effect on the aldehyde to ozonide ratio obtained from NMR spectra, especially in the ozonation of neat SFO.
The aldehyde to ozonide ratios obtained regardless of the extent of ozonation were 10.5:89.5 and 46.6:53.4 for SFO ozonized in the absence and presence of water, respectively.
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Comparative Studies of O3 Ozonated Olive Oil and Ozonated Sunflower Oil.
Díaz M., Hernández R., Gómez M., Fernández H., Martínez G., Curtielles V., Vélez H. and Garcés R.
(Cuba & Spain)
Prior studies have proven ozonated vegetable oils are useful for medical purposes in different diseases, taking into account their broad spectrum germicidal power.
In this study the ozonated olive oil and ozonated sunflower oil are compared chemically and microbiologically. Ozone-oxygen gas was bubbling into the two oils until they were solidifies.
The peroxide, acidity and iodine indexes along with antimicrobial activity against Staphylococcus aureus ATCC 6538,Echerichia coli ATCC 10536, Pseudomonas aeruginosa ATCC 27853 and Bacillus subtilis ATCC 6633 were determined.The reaction products were identified using Proton Nuclear Magnetic Resonance (1H NMR).
Ozonation effects on the fatty acid composition of these oils were analyzed using Gas-Liquid Chromatographic Technique (GLC).
- An increase in peroxidation index was observed in both oils but was higher in O3 ozonized olive oil.
- The acidity index was similar in both oils and the iodine value was zero in ozonized olive oil whereas in ozonized sunflower was 10.65 Units, this is indicative that all unsaturated group in olive oil reacted with ozone, but not the same in sunflower oil.
- The highest action spectrum of antimicrobial activity was obtained with the higher peroxide index from both ozonated oils.
- Signals of oxygenated groups through 1H NMR were assigned.
- Composition of fatty acids in both ozonized oils showed gradual decrease in (C18:1, C18:2) unsaturated fatty acids with gradual increase in ozone doses.
- It was concluded that both ozonated oils have similar germicidal power and physical-chemical characteristics.
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Antimicrobial Activity of O3 Ozonated Sunflower Oil on Streptococcus mutans.
Fernández I., Curtiellas V. and Sánchez E.
Streptococcus mutans is considered the main ethiological agent of buccal caries. The aim of this work was to measure the lethality action of ozonated sunflower oil (OLEOZON®) on Streptococcus mutans. The influence of different conditions, like pH, agent concentration and contact time on OLEOZON® lethality was also evaluated. The values of minimum inhibitory concentration (CMI) and minimum bactericide concentration (CMB) were determined.
It was demonstrated that OLEOZON® activity did not variate significantly with the studied variation of pH, although the microorganism number decreases to pH lightly acid. The study of the influence of the different agent concentrations showed a significant effect of OLEOZON® in the microorganism lethality, being 287.5 mg/mL the most effective studied concentration.
An exposition time, between the agent and the microorganism, of ten minutes, produce a 99,9 % of microorganism lethality. However with only three minutes of exposition time was obtained a reduction of 95,5 % of initial microorganism concentration.
These results proved that OLEOZON® is an effective drug to treat Streptococcus mutansinfection.
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Physical Chemical Characteristic of Cosmetic Ceam with O3 Ozonized Theobroma Oil.
Díaz M., Fernández H., Gómez M., Hernández R., Martínez G., Hernández D., Vargas E., Díaz A., Hasbun S. and Ledea O.
Cosmetic creams with active oxygen present moisturizer and conditioner effects. Therefore, skin oxygenation and microcirculation are increased preventing cutaneous aging. In this work an anti-wrinkle cosmetic cream with ozonized theobroma oil is presented.
This cream has been approved for Health Cuban Authorities as anti-wrinkle cosmetic cream with moisturizer and conditioner properties. Chemistry and microbiological stabilities, physical-chemistry properties, rheology and skin and ophthalmic irritably test were performed.
The cream is stable for six months. Peroxide values was minor that 65 mmol-equiv O 2 active/kg of sample; acidity was minor that 15 Units, pH values between 4 and 8, and visual evaluation as homogenous consistent solid of white colour. The behaviour of fluidity graphic was plastic with tixotropic value of 14 740.
The tests of irritably of the cream were negative. It was concluded, the cream with ozonized theobroma oil maintains their quality parameters during six mounts from be using in cosmetology.
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Physical-Chemical Aspects of O3 Ozonized Sunflower Oil
Hernández R., Martínez G. and Díaz M.
The knowledge of the physico-chemical properties of ozonized vegetable oils has a great importance for its characterization and identification. In this study a correlation of the relative density and viscosity with the peroxides index of the ozonized sunflower oil is carried out. Furthermore, main formed oxygenated products are identified after the ozonization by means of Proton Nuclear Magnetic Resonance (1H-NMR) signals.
The peroxides index and the relative density showed a linear relationship with a correlation coefficient of 0.9991 and a determination coefficient of 99.82 %, while the peroxide index and the viscosity showed an exponential relationship with a determination coefficient of 98.52 %. An increase in peroxidation index, in viscosity and oxygenated compounds concentration were observed during the course of ozonization reaction.
These results suggest that physical-chemical properties can provide valuable information about of ozonization grade of sunflower oil. This study could also be the base of future chemical investigations of ozonized vegetable oils.
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Measurement of Hydroperoxides in O3 Ozonated Sunflower Oil.
Martínez G., Hernández R. and Díaz M.
Total hydroperoxides (HPT) concentration using the ferrous oxidation in xylenol orange (OFX) assay and peroxide value (IP) using iodometric assay, were determined in sunflower oil samples ozonated at different ozone dosage.
Effect of increasing ozonazed sunflower oil amount was evaluated by assembly assay, where a lineal relationship was obtained between oil amounts from 2 to 17 µg in the assay and absorbance units measured at 560 nm. The OFX reagent calibration and the extinction coefficient calculation were carried out for HPT measurement in ozonazed samples. The correlation established between IP and the concentration obtained by OFX assay showed a lineal relationship (r = 99.29; r2 = 98.59 %).
These results confirm that this assay is appropriate for HPT measurement in ozonazed sunflower oil.
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Variation of O3 OLEOZON®'s Iodometric Peroxide Value with the Measurement Time.
Ledea O., Díaz M., Vargas E., Martínez G., Hernández R., Fernández H. and Hernández D.
The reaction of ozone with unsaturated triglycerides, present in vegetable oils, produces different types of peroxidic compounds as described by Criegee mechanism. The peroxide value is the method generally employed to determine total peroxidic compounds formed.
The United States and British Pharmacopoeias established an iodometric method with a reaction time of two minutes to analyze the samples.
Usually, the samples of oils and fats present a low content of peroxidic compounds, but OLEOZON® contains a great amount and diversity of them, and certainly higher reaction times could be necessary to determine its total peroxide content.
In this work, the variation of iodometric peroxide value of OLEOZON® with the reaction time was evaluated. A representative sample of ten batches of the drugs were randomized chosen for the study, and the reaction times assayed were: 2 min, 4 h, 8 h, 16 h, and 24 h.
The peroxide value of OLEOZON® increases with the reaction time and the increment was determined by the history of each batch.
The influence of storage time, viscosity, and peroxide value measured at two minutes reaction time, on peroxide value measured at higher reaction time were evaluated.
In general, the variation coefficient of the determination, decrease at the same time that the reaction time increase.
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Nuclear Magnetic Resonance Spectroscopic Characterization of O3 Ozonated Sunflower Oil.
Díaz M., Gavín J., Ledea O., Hernández F., Alaiz M. and Garcés R. (Cuba & Spain)
Ozonation reactions are very important in vegetable oil chemistry since their ozonation products are involved in antimicrobial effect in therapeutical uses for several microbiological etiology diseases.
Information on the spectroscopic characterization of the products generated by ozonolysis of sunflower oil is limited. In the present study ozonized sunflower oil with 650 mmol-equiv/g of peroxide index is chemically characterized.
Ozonation of sunflower oil produced ozonides, aldehydes and hydroperoxides which were identified by 1H, 13C and two-dimensional 1H Nuclear Magnetic Resonance (NMR).
The virgin sunflower oil and ozonized sunflower oil show very similar 1H NMR spectra except for the resonances at d = 9.74 and d = 9.63 ppm that correspond to both triplet from aldehydic protons, d = 5.6 ppm (olefinic signal from hydroperoxides), and d = 5.15 ppm (multiplet from ozonides methylic protons).
Other resonance assignments are based on the connectivities provided by the proton scalar coupling constants. These are the following: d = 3.15 ppm (doublet from methylenic group in a position respect to olefinic proton), d = 2.45 ppm (multiplet from methylenic group allylic to ozonides methynic protons) and d = 1.62 ppm (multiplet methylenic protons in b position respect to ozonides methynic protons).
From the 13C NMR and 1H- 13C two-dimensional spectrum of the ozonized sunflower oil, the presence of ozonides was confirmed by the signals d = 103.43 and d = 103.49 ppm respectively. The others new signals found in d = 42.5 and d = 42.76 ppm confirm the presence of methylenic carbons from hydroperoxides and ozonides.
These results indicate that the NMR Spectroscopy can provide valuable information about the amount of reaction compounds of ozonized vegetable oil. From the chemical structural elucidation of ozonated sunflower oils, relevant biochemical and chemical information can be achieved.
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Identification of O3 OLEOZON® Main Peroxidic Compounds.
Ledea O., Rosado A., Molerio J., Besada V., Betancourt L., Díaz M., González L., Tambara Y. and Hernández C.
The ozonized sunflower oil for topical applications (OLEOZON®) is a registered drug obtained through the reaction between ozone and sunflower oil according to a process developed at Ozone Research Center. OLEOZON® has shown antimicrobial effects against virus, bacteria, protozoa, and fungi, but its mechanism of action remains still unknown.
The germicide activity of the ozonized vegetable oils has been usually associated to peroxidic compounds formed by the reaction of ozone with unsaturated triglycerides. However, the chemical information available in the specialized literature about the structure of these peroxidic compounds is very limited.
In the present work, the electrospray mass spectrometry technique was used to analyze the main peroxidic compounds present in the OLEOZON®. 1H and 13C Nuclear Magnetic Resonance and Infrared Spectroscopy were also employed. Peracids, a -hydroxy-hydroperoxides, a -acyloxy-hydroperoxides and ozonides, were identified as the main peroxidic compounds in OLEOZON®.
This is the first time than the electrospray mass spectrometry is used to analyze the peroxidic and polymeric compounds present in ozonated or oxidized vegetable oils. Also, the presence of peracids in the ozonated vegetable has not been reported previously and a mechanism for the formation of this peroxidic compound was proposed.
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Antibacterial Activity of Topic O3 OLEOZON®; Packed in glass and Polyethylene Flasks.
Gómez M., Curtiellas V., Fernández I., Sánchez E., Fernández H. and Díaz M.
Ozonized sunflower oil (Topic OLEOZON®) presents a high germicidal power. This product is a wide spectrum antimicrobial agent of with inhibitory and lethal activity on bacterial, protozoa and fungi.
The aim of this work was to study the microbiological activity from topic OLEOZON® stored to temperatures from 2 - 8 oC. Escherichia coli ATCC 10536,Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 6538 and Bacillus subtilis ATCC 6633 were selected as test organism.
Topic OLEOZON® was packed in three types of containers like amber colour glass, density low polyethylene and density high polyethylene flasks. Biological evaluation of product was done by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) based on National Committee for Clinical Laboratory Standards (NCCLS) guidelines, for each flask type and storage period (0, 2, 5, 7 and 14 months).
Topic OLEOZON® was effective against all strains tested.
MIC values showed an increase of 5 mg/mL to 13 mg/mL that corresponding to 0 and 14 months, MBC values also showed an increase from 30 mg/mL to 148 mg/mL from 0 to 14 months.
Taking into account that this product is directly applied on the wounded skin at a concentration near to 950 mg/mL, these results show that Topic OLEOZON® has high biological activities even when it is stored during fourteen months from 2 - 8 oC in all flasks type studied.
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New quality system in the Ozone Research Center.
Nicolás P.D., Díaz M., Fernández H., Hernández R., Martínez G., Vargas E., Hernández D., Mora C., Curtiellas V. and Rodríguez M.
Ozonized sunflower oil OLEOZON® is a wide spectrum antimicrobial product with inhibitory and lethal activity on grampositive and gramnegative bacterias.
This product, during several years, has been elaborated in the pilot plant of Ozone Research Center. OLEOZON® has approved the Medical Registry for the Epidermofitosis disease, pathology of fungicide origin, very frequent in many countries. Also, their production process has been validated according to the norms settled down by the competent health sanitary authorities (CECMED).
The new Quality Administration System is conformed by Administration Unit of the Quality that includes following areas: Quality Insurance, Quality Control, Production, Maintenance, Administrative and Security. All these areas are command by Quality Insurance area and the whole System is take command by Center Direction. The codes of the Normalized Procedures of Operation (P.N.O) were changed.
In general the system has 166 P.N.O among modified and new. The following support programs were created: Maintenance Program, Calibration and Verification, Hygienic Program, and the modified Training Program. The new Quality Administration System was implanted starting from May of 2003 and the License of Production Renovation was obtained.
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