Spectroscopic Characterization of Ozonated Sunflower Oil

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/kg 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 δ = 9.74 and δ = 9.63 ppm that correspond to both triplet from aldehydic protons, δ = 5.6 ppm (olefinic signal from hydroperoxides), and δ = 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: δ = 3.15 ppm (doublet from methylenic group in α position respect to olefinic proton), δ = 2.45 ppm (multiplet from methylenic group allylic to ozonides methynic protons) and δ = 1.62 ppm (multiplet methylenic protons in β 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 δ = 103.43 and δ = 103.49 ppm, respectively. The others new signals found in δ = 42.5 and δ = 42.76 ppm confirm the presence of methylenic carbons from hydroperoxides and ozonides. These results indicate that 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.     

1H NMR Study of Methyl Linoleate Ozonation.

Unsaturated fatty acids are essential components of vegetable oils and cellular membranes and the involved aspect of unsaturated fatty acids ozonation have been widely studied by different authors. In this paper, in vitro ozonolysis of unsaturated fatty acids with addition of water or ethanol has been studied by Proton Nuclear Magnetic Resonance (¹H NMR) at 250?MHz in order to explore the possibility of this technique for the detection of Criegee ozonides in characterizing ozone reaction with these substrates. The ozonolysis of methyl linoleate showed that signal intensities from formed ozonides were increased with ozone concentration increments. However, the signal intensities with addition of water were higher than those in ethanol addition. Signal intensities from olefinic double bonds were found to decrease with the increment in ozonide signals. Thus, a correspondence of the behavior of these signals is observed with a proportional rate reaction between the number of double bonds in the substrate molecule. Signals from aldehyde formation were poorly detected at lower ozone concentration. It was concluded that the evaluation of ozonide and olefinic double bond signals from 250?MHz ¹H NMR can be a useful tool in assessing ozone reaction with biomolecules. The reaction mechanism for the ozone reaction with unsaturated fatty acids in the presence of water or ethanol is analyzed.     

Study of Three Systems of Ozonized Coconut Oil

In the present study, the products from the ozonation of three systems (ozonized coconut oil, ozonized coconut oil with water and ozonized coconut oil with ethanol) are characterized and compared statistically. Peroxide, acidity and viscosity indexes were determined. The reaction products were identified by ¹H NMR and their antimicrobial activity was evaluated. The ozonized coconut oil with ethanol showed the highest peroxide and acidity indexes. This result suggests that in presence of ethanol a greater peroxide decomposition occurs leading to greater acid formation. The variation coefficients obtained in the analysis methods were smaller than 10%. Reaction products were identified as ozonides and aldehydes compounds. The highest action spectrum of antimicrobial activity by Staphylococcus aureus was obtained with the ozonized coconut oil with water and ethanol systems.     

Germicidal Properties of Ozonated Sunflower Oil

The aim of this work was to establish the minimal inhibition concentration (MIC) of ozonated sunflower oil for various microorganisms. To determine the influence of the ozonated medium on the growth of bacteria Bacillus subtilis, Escherichia coli and yeast Candida albicans the Petri dish method was used. Chemical and physical properties of ozonated sunflower oil were additionally studied. Microbiological studies proved that these microbes have various sensibility against ozonated oil. The most resistant are gram-negative bacteria E. coli and the yeast C. albicans. Gram-positive bacteria, B. subtilis turned out to be less resistant, because no growth was observed for preparation with an ozone dose of 200 mgO₃/g oil.     

Study of Ozonated Olive Oil: Monitoring of the Ozone Absorption and Analysis of the Obtained Functional Groups

The peculiarities of ozone absorption during olive oil ozonolysis have been studied by continuous monitoring of ozone concentrations at the bubbling reactor outlet. The determined amount of ozone, consumed during the ozonolysis of the double bonds, was used as alternative way for evaluation of the degree of unsaturation of the oil. The basic functional groups, products of the reaction: ozonides and aldehydes have been quantitatively characterized by means of ¹H-NMR spectroscopy, whereupon their ratio was found to be 93.4:6.6 (mol. %), respectively. The determined ratio between the cis and trans ozonides was 46:54.     

A New Formulation Based on Ozonated Sunflower Seed Oil: In Vitro Antibacterial and Safety Evaluation

The objective of this study was to investigate the potential in vitro biological properties of Oz.Or.Oil 30, a new formulation composed of 30% ozonated sunflower seed oil, which is believed to keep skin smooth and moisturized, supporting repair processes, tissue regeneration and re-epithelialization of wounds. The antibacterial activity, the qualitative and quantitative evaluation of the cytotoxic effect of the formulation on cultures of Vero cells and 3T3 fibroblasts showed that Oz.Or.Oil 30 merits further in vivo study using clinical-laboratory correlations, because it could be suggested as an alternative therapy against bacterial and fungal diseases.     

Stability Studies of Ozonized Sunflower Oil and Enriched Cosmetics with a Dedicated Peroxide Value Determination

Ozonized oils have interesting applications in the cosmetic industry and several patents on enriched products were developed in the last years. Ozonides are known for their high reactivity and stability data are required for enriched cosmetic products during storage. In this paper a dedicated determination of Peroxide Value (PV) was performed on ozonized sunflower oil (Neozone 4000) and on some enriched cosmetics, in order to control their stability during storage. The optimal conditions of the PV method were determined, following the decomposition reaction of ozonides with KI utilizing ¹H-NMR and GC/MS techniques.     

Measurement of Peroxidic Species in Ozonized Sunflower Oil

Peroxidic species in ozonized sunflower oil using different methods as iodometric and ferrous oxidation in xylenol orange (FOX) were measured. The necessary reaction time from two minutes up to 36 hours using iodometric assay in ozonized sunflower oil was determined. Peroxide values achieved maximum values at 24 hours of reaction time. Hydroperoxides content measured by FOX assay and peroxide value determined at two minutes using iodometric assay had a linear relationship (r² = 98.18%), while, at 24 hours a logarithmic relationship (r² = 98.39%) was shown. Values of hydroperoxides were lower than peroxides values at 24 hours and represent between 23 and 44% in all samples of ozonized sunflower oil studied.     

Quantitative triacylglycerol analysis of whole vegetable seeds by1H and13C magic angle sample spinning NMR spectroscopy

High-resolution¹³C and¹H magic angle sample spinning nuclear magnetic resonance (NMR) spectra have been obtained and used to define the relative unsaturated acyl distribution of triacylglycerols in whole oil seeds. Inverse gated proton decoupled¹³C and¹H NMR spectra permit the quantitative analysis of seeds containing simple oils,e.g., sunflower seeds containing oleyl and linoleyl unsaturates only. More sensitive¹³C NMR techniques are necessary for the analysis of specific seed classes. One such class is the rapeseed, which is especially difficult due to its low oil content (≈ 2 mg oil/seed) and complex unsaturated acyl profile of oleyl, linoleyl, linolenyl, erucyl, and eicosenoyl. The Distortionless Enhancement by Polarization Transfer technique significantly improves sensitivity to the extent that single rapeseeds can be examined within an hour of acquisition time. Furthermore, some positional (1,3- or 2-glycerol attachment) groups can be identified leading to a partial estimation of the 1,3-, 2-acyl distribution.     

Ozonated Mineral Oil: Preparation,Characterization and Evaluation of the Microbicidal Activity

The ozonation of vegetable oils has been studied, since the produced ointments have antibacterial and fungicidal activities. However, the ozonation of mineral oils has not been reported in the literature yet, opening an interesting field for examination. In this work, we have shown the ozonation of a commercial mineral oil (Nujol). The main goal was to produce oils containing free ozone, so that this gas could act as antimicrobial agent. It was found that in Nujol, ozone remains for at least 40 days and in the sample ozonized for 15 h its concentration was 7.5 mg mL^?1. Neat Nujol showed no antimicrobial activity against the tested microorganisms, however, when ozonated it showed antimicrobial activity against Enterococcus faecalis, Staphylococcus aureus and Escherichia coli.     

Acyl Migration Kinetics of 2-Monoacylglycerols from Soybean Oil via <Superscript>1</Superscript>H NMR

The acyl migration kinetics of neat 2-monoacylglycerol (2-MAG) to form 1-MAG was determined using ¹H NMR spectroscopy to monitor the β-proton integration ratios of the two species over time. 2-MAG was synthesized by the Novozym 435-catalyzed alcoholysis of soybean oil and isolated by solvent extraction or molecular distillation at a mole fraction (X _2-MAG) of 0.94 relative to total MAG. The kinetics parameters of the neat 2-MAG acyl migration were investigated over the temperature range of 23–80 °C. The 2-MAG mol fraction remained unchanged at 23 °C over the course of 168 h and reached an equilibrium of X _2-MAG = 0.09 at only 80 °C. Modeling of the kinetics data revealed a 2-MAG half life (t _1/2) of 3,500 and 22.8 h at 23 and 80 °C, respectively, with an activation energy of 79.0 ± 6.5 kJ mol⁻¹. The use of ¹H NMR spectroscopy proved an expedient method for monitoring the acyl migration in 2-MAG compared to other reported methods (e.g. GC, HPLC, and ¹³C-NMR spectroscopy), requiring no sample manipulation and minimizing the deleterious effects of high temperatures and solvent exposure. Product names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product to the exclusion of others that may be suitable.     

High-resolution F and H NMR of a vinylidenefluoride telomer

The reaction products of vinylidenefluoride (VDF) with methanol as a telogen have been analysed in the solution state by ¹H and ¹⁹F nuclear magnetic resonance (NMR) spectroscopy. High-resolution ¹⁹F and ¹H NMR spectra were achieved using high-power ¹H and ¹⁹F decoupling, respectively, giving superior resolution and revealing previously unresolved signals of the vinylidenefluoride telomer (VDFT). ¹H and ¹⁹F homo- and hetero-nuclear scalar coupling constants are presented and the spectra of functional groups and reverse units (including the identification of short-chain structures) are discussed. Furthermore, the application of ¹⁹F or ¹H decoupling for the correct assessment of reverse-unit content and degree of polymerisation is demonstrated. This work highlights the need for high-resolution NMR spectroscopy to determine both the chemical structure and the composition of these important fluoropolymers.     

Effect of α-Tocopherol During In Vitro Ozonation of Methyl Linoleate: Its Implication in Ozone Therapy

Systemic ozone therapy is widely used as an oxidant therapy to treat many conditions and diseases. It is known that ozone therapy acts through a transient oxidative stress produced by lipid ozonation products. The convenience of supporting patients with antioxidants during systemic ozone therapy applications is now under discussion. We studied the reaction of linoleate (one of the main constituents of cellular membranes and plasma phospholipids) with ozone in presence or absence of α-tocopherol, in order to explore whether the combination of ozone and antioxidant has some effect on fatty acid ozonation products. Proton nuclear magnetic resonance spectroscopy (¹H NMR) was used for following the reaction between 4.8mol mmol of methyl linoleate and 2.08 or 4.32mmol of ozone, with addition of different amounts of α-tocopherol (0.10, 0.18, and 0.26μmol). Ozonide (δ=5.15ppm) and aldehydes (δ=9.63ppm and δ=9.74ppm) intensities from ¹H NMR signals markedly decreased with α-tocopherol addition. When αtocopherol is absent, the intensities from olefinic proton signals diminished with ozone concentration increment; however, with αtocopherol in the mixture a smaller decrement was achieved. No detectable signals were found with the ozonation of α-tocopherol without methyl linoleate in the reaction mixture. These results suggest that α-tocopherol reacts with those products released from the reaction of ozone with methyl linoleate. This fact points out that antioxidant supplementation during systemic ozone therapy (major and minor autohemotherapy, rectal insufflation, and so forth) can be detrimental toward achieving the needed transient oxidative stress responsible for biological activities.     

Antifungal Activity of Olive Oil and Ozonated Olive Oil Against Candida Spp. and Saprochaete Spp.

Ozonated olive oil was investigated for their capacity to inhibit growth of 38 yeast strains of Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Saprochaete capitata. Two different ozonated olive oil (OZO1, OZO2) and two different olive oil (OL1, OL2) samples having different biochemical parameters were assessed in terms of their antifungal ability and comparison was made. Fluconazole was chosen as control antifungal agent. Each sample’s antifungal activity decreased in the following order: OZO1 > OZO2 > OL1 ≥ OL2. This study demonstrated that ozonated olive oil may help to control some fluconazole-resistant and dose-dependent sensitive fungal strains.     

Study on relationship between the concentration of hydrocarbon groups in heavy oils and their structural parameter from H NMR spectra

More than 60 heavy oils samples, belonging to the hydrogen adding tail oil, catalytic oil slurry, catalytic heavy oil slurry and catalytic heavy tar waxy oil, which taken from different processing units in Nanjing Refinery, are analysed by ¹H NMR and column chromatography. The analytical data of the heavy oils by two methods are compared and correlated, while good relationships between the specific area proton's percentage in ¹H NMR spectra and the contents of paraffinic and alicyclic hydrocarbons ((P+N)%), aromatic hydrocarbons (A%) are found. Our ¹H NMR method in calculating the amount of (P+N) and A in heavy oils proved fast, convenient and reliable.     

Gasoline composition determined by H NMR spectroscopy

A fast and simple ¹H NMR spectroscopic method was developed for the concentration measurement of aromatic, olefinic and aliphatic fractions in gasoline samples. Simultaneously, individual components such as benzene, methyl tert-butyl ether and several dienes could be determined. The method relies on only a few, well-established assumptions about the molecular compositions and is therefore applicable to a broad range of gasolines. It is well suited for laboratories with high sample throughput since measurement time is short and all concentrations are determined within one NMR experiment. The method was applied to commercial gasolines and samples used in European round robin tests. Comparisons of NMR and round robin test results showed excellent agreement.     

A simple method for determining protic end-groups of synthetic polymers by H NMR spectroscopy

A simple method for the determination of protic end-groups (-XH) in synthetic polymers involves in situ derivatization with trichloroacetyl isocyanate (TAI) in an NMR tube and observation of the imidic hydrogens of the derivatized products [-X-C(O)-NH-COCCl₃] by ¹H NMR spectroscopy. In this paper, we report that the method is effective for the quantitative determination of hydroxy, primary amino and carboxy end-groups of polymers with . It may also be applied to detect chain ends in higher molecular weight polymers. The signals for the imidic (and, in the case of amines, amidic) hydrogens appear in a region (δ 7.5-11) that is clear of other signals in the case of most aliphatic polymers and many aromatic polymers such as polystyrene and poly(ethylene terephthalate). The method has been applied in the characterization of polymers formed by conventional and living radical polymerization (RAFT, ATRP, NMP), to end functional poly(ethylene oxide) and to polyethylene-block-poly(ethylene oxide). The method appears less effective in the case of sulfanyl end-groups. The chemical shift of the imidic hydrogen shows remarkable sensitivity to the microenvironment of chain end. Thus, the imidic hydrogens of TAI derivatized polyethylene-block-poly(ethylene oxide) [PE-(EO)_mOC(O)NHC(O)CCl₃] are at least partially resolved for m=0, 1, 2, 3 and ≧4 in the 400 MHz ¹H NMR spectrum. It is also sensitive to the chain end tacticity of, for example, amino-end-functional polystyrenes and thus to the relative configuration of groups removed from the chain-end by two or more monomer units. TAI derivatization also facilitates analysis of amine functional polymers by gel permeation chromatography (GPC) which is often rendered difficult by specific interactions between the amine group and the GPC column packing.     

Effect of time on the hydration and temperature-induced phase separation in aqueous polymer solutions. H NMR study

Dehydration during temperature-induced phase separation in D₂O solutions of poly(vinyl methyl ether) (PVME), poly(N-isopropylmethacrylamide) (PIPMAm) and poly(N-isopropylacrylamide) (PIPAAm) was followed from time dependences of NMR spin-spin relaxation times T₂ of HDO. Both the time characterizing the exclusion of the water from mesoglobules (manifested by the increase in T₂ values) and the induction period which precedes the increase in T₂ values, increased in the order PVME < PIPMAm < PIPAAm. For D₂O solutions of PIPMAm/PVME (or PIPMAm/PIPAAm) mixtures a direct connection between the state of the mesoglobules (hydrated or dehydrated) formed by the component with lower LCST (PVME, PIPAAm) and the temperatures of the phase transition of the PIPMAm component was established by NMR spectroscopy.     

Quantification of soybean oil ethanolysis with <Superscript>1</Superscript>H NMR

In this study, proton NMR spectroscopy (200 MHz) was used for quantifying the content of ethyl esters in known mixtures of soybean oil and ethyl soyate (biodiesel). For this purpose, the peak areas of ester ethoxy and glycerol methylenic peaks in the region of 4.05–4.40 ppm were measured and a calibration plot of the respective peak areas vs. the known composition of the oil/ethyl ester mixtures was used. The transesterification values determined in this way were compared with viscosity and total glycerol determinations and a good correlation was obtained. Therefore, for routine analysis, the conversion (in %) of oil to ethyl esters was determined. The methodology presented in this work proved to be quicker and simpler than others reported in the literature, such as GC and/or HPLC.     

The use of <Superscript>1</Superscript>H NMR for yield determination in the regioselective epoxidation of squalene

¹H NMR can be used to determine the epoxide yield rapidly in the oxidation of squalene. Moreover, unequivocal distinction can be made between internal and terminal epoxide bonds. To underline the power of this technique, different stoichiometric and catalytic epoxidation procedures were carried out using squalene as substrate. They were characterized in terms of substrate conversion and regioselectivity of the epoxide fraction.