Quantitation and distribution of altered fatty acids in frying fats

The distribution and quantity of polar compounds and altered fatty acids in used frying oils, collected by Food Inspection Services of the Junta de Andalucía in Spain, was measured. Additional samples evaluated were sunflower oil, high-oleic sunflower oil, and palm olein that were subjected to thermoxidation and frying in laboratory experiments. A combination of adsorption and high-performance size-exclusion chromatography was applied to the oil samples both before and after transesterification. Through analysis of fatty acid methyl ester derivatives, differentiation of four groups of altered fatty acids (oxidized monomers, nonpolar dimers, oxidized dimers, and polymers) could be attained. Evaluation of real frying samples with polar compound levels around the limit for fat rejection (21.1–27.6% polar compounds) gave values of total altered fatty acids ranging from 8.1 to 11.3%, and levels higher than 20% were found in the most degradated samples. The results obtained clearly support the need for control and improvement of the quality of used fats in fried-food outlets.     

Enzymatic hydrolysis of fractionated products from oils thermally oxidized in the laboratory

Enzymatic hydrolysis of the acylglycerol products obtained from thermally oxidized vegetable oils was studied. Corn, sunflower and soybean oils were heated in the laboratory at 180 C for 50, 70 and 100 hr with aeration and directly fractionated by silicic acid column chromatography. By successive elution with 20%, then 60% isopropyl ether inn-hexane, and diethyl ether, the thermally oxidized oils were separated into three fractions: the nonpolar fraction (monomeric compounds), slightly polar fraction (dimeric compounds), and polar fraction comprising oligomeric compounds. Enzymatic hydrolysis with pancreatic lipase showed that the monomers were hydrolyzed as rapidly as the corresponding unheated oils, the dimers much more slowly, and the oligomeric compounds barely at all. Overall, the hydrolysis of the dimers was less than 23% of that for the monomers, with small differences among the oils. Longer heating periods resulted in greater reductions in hydrolysis of the dimeric compounds. These results suggest that the degree of enzymatic hydrolysis of the fractionated acylglycerol compounds is related to differences in the thermal oxidative deterioration, and amounts of polar compounds in the products.     

Hydrolysis of fish oils containing polymers of triacylglycerols by pancreatic lipasein vitro

Fish oils containing different levels of polymers of triacylglycerols formed during autoxidation were incubated with pancreatic lipase to establish whether these polymers are substrates for lipase hydrolysis. With oils containing low amounts (less than 4%) of triacylglycerol polymers as substrates, both triacylglycerols and polymers of triacylglycerols were almost completely hydrolyzed, and fatty acid monomers and monoacylglycerols were the major lipid products. Under the same incubation conditions, some triacylglycerols remained intact when highly oxidized oils containing 20 or 30% triacylglycerol polymers were the substrate. The fatty acid composition of these residual triacylglycerols was almost identical to that of triacylglycerols present at the start of the assay. When fish oil containing 30% triacylglycerol polymers was incubated with the lipase, the component triacylglycerols and polymers of triacylglycerols were hydrolyzed at similar rates, and fatty acid dimers were detected as a product. It is concluded that the high molecular weight polymers of triacylglycerols present in oxidized fish oils can be hydrolyzed by pancreatic lipasein vitro.     

Lymphatic absorption of nonvolatile oxidation products of heated oils in the rat

The lymphatic absorption of nonvolatile oxidation products (NVOP) formed during heating of fats was studied. Heated colza or soybean oils or synthetic triglycerides containing a definite aromatic or alicyclic fatty acid were fed to thoracic duct-cannulated rats. Tritium-labeled triolein was added to each dietary fat, as an internal standard, in order to calculate the percentage of lymphatic absorption of the ingested NVOP. Results show that 4% of the total polymeric acids, 53% of the total oxidized monomeric acids and 96% of the total cyclic monomeric acids were recovered in the lymphatic lipids. Gas liquid and quantitative thin layer chromatography of these 3 classes indicated that, within a NVOP class, the various constituents did not present the same absorption rate. The lymphatic absorptions of individual oxidized monomers were between 25 and 93%. Concerning the polymer fraction, the lymphatic recoveries were 1% (nonpolar dimers), 6.8% (polar dimers) and 12% (polar oligomers). Aromatic acids were absorbed to a lesser degree (50–60%) than cyclohexenic acids (91–98%).     

Absorption in rats of rapeseed, soybean, and sunflower oils before and following moderate heating

Rapeseed, soybean, and sunflower oil were heated for 15 min in a 5-mm oil layer in a pan at 180°C. The fatty acid composition was almost unaffected by heating, while the polymer content rose slightly and the tocopherol content decreased, except in soybean oil. The absorption of oils before and after heating was investigated in lymph-cannulated rats. Oils were administered as emulsions through a gastrostomy tube and lymph was collected during the next 24 h. The highest accumulated lymphatic transport of total fatty acids was observed after administration of rapeseed oil, and the lowest after heated sunflower oil. The accumulated transport was similar for all unheated oils. The transport of fatty acids was significantly lower in rats receiving heated oil compared to those receiving the corresponding unheated oil. Small increases in polymers may have contributed to the decreased lymphatic transport of oil following heating, although this probably does not fully explain the effect. The absorption of sunflower oil was more affected by heating than the absorption of soybean or rapeseed oil. Furthermore, the largest decrease in total activity of tocopherols following heating was observed in sunflower oil. Overall, these results demonstrate that the absorption of vegetable oils is affected by moderate heating.     

Long-term responses of rats to heat-treated dietary fats: IV. Weight gains,food and energy efficiencies,longevity and histopathology

Representative cottonseed salad oils, corn oils, lards and hydrogenated vegetable shortenings, and portions of the same fats heated at 182 C for 120 hr were fed as 20% of nutritionally adequate diets to weanling albino rats in longevity studies. Differences in the responses of rats fed diets containing the unheated and heated fats were generally small with respect to rates of gain, 12 th week and adult weights, efficiencies of utilization of absorbed energy, incidences of grossly detectable diseases and longevities. There were no indications that the feeding of the heated fats had shortened survival times in comparison with the comparable unheated fat. Animals fed hydrogenated vegetable shortening, heated or unheated, survived the longest. However, gains were slightly slower with the heated cotton-seed oil diets, and food efficiencies were slightly lower with the heated cottonseed oil and heated lard diets because of decreased digestibilities of these fats. The usual disabilities of old age such as nephritis, respiratory disease and periarteritis were present in all groups. The incidence of mammary tumors was high but did not differ significantly with the kind of fat, heaated or unheated. Tumor incidence other than mammary was similar in both sexes and there was no significant difference between fresh and heated fats. Absence of adverse effects attributable to the heated fats during the life span of the rats in further evidence of the safety of these fats of the quality customarily consumed by the human population.     

Applications of chromatographic techniques to evaluate enzymatic hydrolysis of oxidized and polymeric triglycerides by pancreatic lipase in vitro

With the aim of studying the susceptibility to enzymatic hydrolysis of oxidized and polymeric triglycerides (TG) that are formed during frying, various chromatographic techniques were applied in combination, i.e., adsorption chromatography, high-performance size-exclusion chromatography (HPSEC), and thin-layer chromatography-flame ionization detection (TLC-FID). Polar fractions, isolated by adsorption chromatography from thermoxidized trilinolein as model system, and real used frying fats and oils, were analyzed by HPSEC before and after incubation with pancreatic lipase in vitro. Also, the influence of degradation level of used frying oils on hydrolysis of intact TG was investigated with the additional aid of TLC-FID. Results showed the high hydrolysis rate of oxidized TG monomers in contrast to the significant discrimination of pancreatic lipase against TG dimers and, particularly, TG polymers. On the other hand, hydrolysis of intact TG can be affected by the presence of dimers and polymers in abused frying oils.     

Oxidative stability of sunflower oils differing in unsaturation degree during long-term storage at room temperature

The objective of this work was to study the evolution of oxidation in sunflower oils differing in unsaturation degree during long-term storage at room temperature. For this purpose, a combination of adsorption and size-exclusion chromatographies was used for quantification of oxidized triacylglycerol (TG) monomers, dimers, and polymers. Conventional sunflower oil, genetically modified high-oleic sunflower oil, and a 1∶1 mixture of the two were used. Results showed that oxidized TG monomers were the only group of oxidation compounds increasing during the early oxidation stage, and an excellent correlation was found between amounts of oxidized TG monomers and PV during the induction period, independently of the degree of oil unsaturation. Both the rate of formation and the amount of oxidized TG monomers accumulated at the end of the induction period increased as the unsaturation degree of the oils tested was higher. The end of the induction period was marked by the initiation of polymerization and exhaustion of tocopherol. Therefore, the concomitant determination of oxidized TG monomers and polymerization compounds provided a complete picture of the oxidation process.     

Enzymatic hydrolysis in vitro of thermally oxidized sunflower oil

The hydrolysis of thermally oxidized sunflower oil by pancreatic lipase was studied in relation to chemical changes in the acylglycerols. Four classes of compounds (monomers, dimers, trimers and polymers) formed from the acylglycerols were separated from the heated oils by column chromatography on silica gel, and further verified by thin layer chromatography. Each fraction, after analyses for generaly properties, was subjected to a time course study of hydrolysis by pancreatic lipase over a 30-min period. After 70 hr of heating, the amount of hydrolysis for the acylglycerol dimers was only about half that of the monomers, and that for the trimers was, in turn, about one-third that of the monomers. The polymers were the least hydrolyzed and showed no further reaction after 5 min. The reduction in enzymatic hydrolysis of isolated fractions from the thermally oxidized oils indicates structural differences, related to formation of polar compounds and polymerization products. Adverse effects on animals from feeding these materials can be attributed partly to inhibition of hydrolysis resulting in less available energy.     

Thermoxidation of substrate models and their behavior during hydrolysis by porcine pancreatic lipase

The behavior of thermoxidized triacylglycerols during hydrolysis catalyzed by porcine pancreatic lipase was evaluated using nonpolar triacylglycerols isolated from palm olein (NPTPO), triolein, and sn-1,3 diolein substrates. Substrates were thermoxidized at 180°C for 1 to 4 h. Owing to formation of polymers and dimers of triacylglycerols, the molecular weight of the thermoxidized substrates increased. After 1 h heating, the concentration of polymers and dimers was similar for the sn-1-3 diolein and triolein samples but higher in NPTPO samples. Conjugated double bonds were formed in all samples, and α,β-unsaturated carbonyl compounds developed through allylic oxidations. These caused increased ultraviolet absorbance at 232 nm. The hydrolysis of heated and unheated samples by the lipase can be described by a Michaelian equation. The enzyme showed a higher apparent V _max and K _M with heated sn-1,3 diolein and triolein than with their unheated counterparts. This was due to the generation of polar compounds which acted as emulsifiers and which favored the formation of an oil/water microemulsion. This behavior was not observed in NPTPO, where heating decreased the apparent V _max and K _M over the first 2 h. Later, a tendency to increase these values was observed. The results could be explained by a balance between concentration of surfactants and of natural emulsifiers in the thermoxidized samples.     

Isolation and characterization of polymers in heated olestra and an olestra/triglyceride blend

High molecular weight components in thermally oxidized olestra (formerly called sucrose polyesters) and a mixture of olestra and soybean oil were characterized. The high molecular weight components of these oils were separated by preparative size exclusion chromatography and analyzed intact by mass spectrometry, infrared, and nuclear magnetic resonance spectroscopy. The materials isolated from the heated olestra were identified as olestra polymers. Materials isolated from the heated mixed oil (olestra and soybean oil) were identified as polymers of olestra and copolymers of olestra and triglycerides. Polymer linkages identified were identical to those resulting from thermal oxidation of natural vegetable oils of similar fatty acid composition.     

Cyclic fatty acids in sunflower oils during frying of frozen foods with oil replenishment

Frying of frozen foods has become popular because it considerably reduces cooking time. Polymers and cyclic fatty acid monomers (CFAM) formed during frying are potentially toxic and therefore their production should be minimized. Twenty discontinuous fryings of different frozen foods were carried out over ten consecutive days, in sunflower oil (SO) and in high‐oleic acid sunflower oil (HOSO), by adding fresh oil after each frying to bring the volume of the fryer oil back to 3 L. CFAM methyl ester derivates were hydrogenated, isolated, concentrated and quantified by HPLC using a reverse‐phase column, followed by gas chromatography. After 20 fryings, significantly higher contents of polar material, polymers and CFAM (all p <0.001) were found in SO than in HOSO. Bicyclic compound formation was four times higher in SO (p <0.001). The fat from the fried potatoes presented a polymer content very similar to that of their corresponding oils. The 100‐g rations of the SO‐fried potatoes from the 20^th frying supply 49 or 15%, respectively, more polymers and CFAM and 1 mg more bicyclic fatty acids than the 100‐g rations of HOSO‐fried potatoes. Because digestion and absorption of polar material, polymers and CFAM occur, the data clearly show the advantageousness and advisability of frying with HOSO rather than SO.     

Nutrition and metabolic studies of methyl esters of dimeric fatty acids in the rat

Methyl esters of dimeric fatty acids were prepared by fractionating a mixture of conjugated linoleic and oleic acids that was heated for 24 hr at 300 C in the absence of air. Rats fed diets containing less than 1% dimers showed no significant difference (P<0.05) in the growth rate, feed efficiency, liver: body weight ratio, and lipid: liver weight ratio from those fed normal diets. A lymph cannulation study using¹⁴C labeled dimers showed that ca. 0.4% of the dimers fed were absorbed within 12 hr and were transported as free acids in the lymph. Within a 28 hr period, 2% of the labeled dimers fed by gastric intubation were oxidized to¹⁴CO₂, and 1% radioactivity was recovered from the urine. The metabolism of methyl oleate appeared normal for rats prefed diets containing dimers.     

Formation and distribution of oxidized fatty acids during deep‐ and pan‐frying of potatoes

The formation of cis‐9,10‐epoxystearate, trans‐9,10‐epoxystearate, cis‐9,10‐epoxyoleate, cis‐12,13‐epoxyoleate, trans‐9,10‐epoxyoleate, trans‐12,13‐epoxyoleate and the co‐eluting 9‐ and 10‐ketostearates during eight successive pan‐ and deep‐frying sessions of pre‐fried potatoes in five different types of vegetable oils – namely cottonseed oil, sunflower oil, vegetable shortening, palm oil and virgin olive oil – was followed and quantified both in fried oils and in fried potatoes by GC/MS after derivatization to methyl esters. These oxidized fatty acids were present at relatively low concentrations in the fresh oils and pre‐fried potatoes while they increased linearly with frying time, reaching up to 1140.8 µg/g in virgin olive oil (VOO) and 186.9 µg/g in potatoes pan‐fried in VOO after eight pan‐frying sessions, with trans‐9,10‐epoxystearate predominating in all cases. The formation of polymerized triacylglycerols (PTG) was also quantified in frying oils by size exclusion HPLC. Pan‐frying caused higher oxidized fatty acid and PTG formation compared to deep‐frying. Epoxyoleates and PTG concentrations were increased after frying in polyunsaturated oils, while epoxystearate and 9‐ and 10‐ketostearate concentrations were increased after frying in monounsaturated oils. No specific absorption of the oxidized fatty acids by the fried potatoes seems to occur. The dietary intake of oxidized fatty acids and PTG by the consumption of fried potatoes was discussed.     

The effect of phytosterol concentration on oxidative stability and thermal polymerization of heated oils

This study determined the effect of adding mixed phytosterols, at various concentrations, on the thermal polymerization and oxidative stability index (OSI) of soybean and high‐oleic sunflower oils. The indigenous tocopherols and phytosterols were removed from the oils by molecular distillation. Pure phytosterols were added back to these stripped oils at concentrations of 0.25, 0.5, 1, and 2.5 wt‐%. These oils were heated at 180 °C, and triacylglycerol dimers and polymers, fatty acid composition, and residual phytosterols were determined. Added phytosterols at 1 and 2.5% significantly decreased thermal polymerization of stripped soybean oil over 8 h. Phytosterols at 2.5% significantly increased polymerization of stripped high‐oleic sunflower oil over 12 h. Added phytosterols did not affect the loss of polyunsaturated fatty acids in either oil. The decomposition of the added phytosterols was followed in both oils during the heating study. The loss of phytosterols in soybean oil ranged from 7 to 13%, while loss in stripped high‐oleic sunflower oil ranged from 13 to 20%. Phytosterols added at 1 and 2.5% significantly decreased the OSI for stripped high‐oleic sunflower oil. This research shows that added phytosterols, especially at higher concentrations, will have an impact on the thermal and oxidative stability of oils.     

Process for production of arachidonic acid concentrate by a strain of mortierella alpina

Various Mortierella alpina fungi were screened for their capacities to produce arachidonic acid. A strain of M. alpina was found to show the highest productivity. Arachidonic acid content of biomass and overall yield per litre of culture was highest in soya flour supplemented medium which produced dispersed mycelium. When the glucose concentration in the medium was varied from 30 to 100 g/L, biomass, lipid, arachidonic acid content of biomass and arachidonic acid yield increased with increasing glucose concentration. Several natural oils, when added to the growth medium, stimulated arachidonic acid production. After fermentation in a 20-L fermenter under optimal culture conditions, the arachidonic acid yield was 5.3 g/L, representing 34.2% w/w of total fatty acids and 13.7% w/w of biomass. An extract containing 72.5% w/w arachidonic acid was prepared from the recovered mycelium.     

Fatty acids based transparent polyurethane films and coatings

Renewable resources used in the preparation of polymers are falling in the class of biomaterials which are rich in hydrocarbons and most of the times obtained from parts of profusely available agriculture feedstock. Use of non-edible vegetable oils and their fatty acids are currently the raw materials focused in synthesizing polymeric materials as renewable sources due to increasing economic and environmental concerns of petroleums. The current oil based polymers are prepared by consuming petroleum based anhydrides and dicarboxylic acids. With this investigation, we tried to substitute almost all components of petroleum feed stock by renewable sources in preparation of polyol. Initially palmitic acid was used as a renewable source to prepare palmitamide by amidation reaction. Further amide was converted into the polyesteramide resin (PEPAD) by reacting it with another renewable resource, i.e. dimer fatty acid which is the dimerized product of unsaturated fatty acids of vegetable oils. Structure of the prepared resin was confirmed by spectroscopic techniques and resin was also characterized by end group analysis like hydroxyl and acid values. The molecular weight of PEPAD was ranged between 2590 and 2760 g mol⁻¹. Palmitic acid based polyesteramide was used to prepare polyurethanes by varying amount of diisocyanate. The transparent polyurethane films and coatings were obtained and examined for water absorption, contact angle, hardness, coating properties, solvent, acid and alkali resistances. The thermal properties of the polyurethane films were estimated by TGA and discussed. It was found that change in NCO content affected on transparency as well flexibility of the prepared films. It also noticed that thermal stability was shifted toward higher range by increasing mole (NCO/OH) ratios due to formation of urea linkages.     

Chemical reactions involved in the deep-fat frying of foods: VIII. Characterization of nonvolatile decomposition products of triolein

Triolein was heated under simulated deep-fat frying conditions at 185 C for 74 hr. The thermally oxidized triolein was converted into methyl esters and then fractionated by urea exclusion. The urea adduct-forming ester (89.2%) was found to be methyl oleate unchanged by the frying treatment. The nonurea adduct-forming esters (10.8%) were further fractionated by silicic acid column chromatography into nine fractions with molecular weights ranging from 304 to 742. Physical and chemical analyses of the fractions indicated that some of them contained oxygen atoms which could not be accounted for by ordinary functional group analyses. The polymers isolated were formed by both carbon-to-carbon and carbon-to-oxygen linkages. The nonpolar dimers were further purified by thin layer and gas chromatography. Structure elucidation revealed that they consisted of a noncyclic dimer and a noncyclic dimer containing a carbonyl group, each of which amounted to 1.36% of the triolein originally used. The polar polymers were studied by depolymerization and the analysis of the depolymerized products. It was estimated that the triolein used for simulated deep fat frying contained 1.1% trimers formed through carbon-to-carbon linkages, 1.9% dimers and trimers joined through carbon-to-carbon linkages, and 3.1% dimers and trimers joined through carbon-to-oxygen or carbon-to-carbon linkages in the same molecule and also dimers and trimers in which all the monomeric units were joined through carbon-to-oxygen linkages. The precise form of the oxygen linkages are not known. However, the fact that it is not cleavaged by HC1 and HI suggests ether linkages.     

Characterization of used frying oils. Part 1: Isolation and identification of compound classes

In this study, an analytical scheme is presented for detailed qualitative comparisons between heated and unheated oils. This scheme is less subject to loss or alteration of sample components when compared with methods that characterize the distillable non-urea adducting portion of heated oils. In this work, oils were first converted to their corresponding fatty acid methyl esters (FAMEs) by transesterification. These FAMEs were then separated by degree of polarity by means of adsorption chromatography and solid-phase extraction. High-resolution capillary gas and liquid chromatography were used to profile isolated fractions. Mass spectrometric and infrared analyses of major chromatographic features were used to identify the presence of aldehydes, epoxides, ketones, alcohols, phytosterols and dimer methyl esters down to 5 ppm in the original sample.     

Improvement of corrosion protection of steel by incorporation of a new phosphonated fatty acid in a phosphorus-containing polymer coating obtained by UV curing

Six formulations containing diacrylate monomers (from 89 to 92.5% (w/w)) as well as a phosphonated methacrylate monomer (from 1 to 10% (w/w)) were prepared. All formulations were UV-cured and the corrosion performance of the resulting coatings applied onto a steel substrate was assessed by electrochemical impedance spectroscopy (EIS). It was first shown that the coatings containing phosphonic acid methacrylate (MAPC1(OH)₂) instead of methacrylate phosphonic dimethyl ester (MAPC1) presented higher corrosion protection related to the strong adhesive properties of phosphonic acid on the metal substrate. A minimum MAPC1(OH)₂ content of 2.5% was determined to provide the highest impedance values (best efficiency). Then, a new bio-based compound, i.e. phosphonic acid-bearing oleic acid (phosphonated fatty acid), was synthesized and added as an inhibitor to the formulations. In the presence of this compound, the corrosion protection was notably improved. The beneficial effect of phosphonated fatty acid was explained by its inhibitive action at the steel/coating interface and by the improvement of the barrier properties.