Volatile components from tristearin heated in air

Tristearin was heated to 192 C in air, and its volatile oxidation products were collected directly on a cooled (−60 C) gas liquid chromatography column. Subsequently, the volatile products were separated by temperature programing up to 250 C and identified by mass spectrometry. Methyl ketones and aldehydes were the major degradation products along with minor amounts of monobasic acids, n-hydrocarbons, primary alcohols, and γ-lactones. Qualitative results indicated all the fatty acid methylene carbon atoms are susceptible to oxidation. Quantities of aldehydes and ketones were found to be in excess of their taste threshold concentrations, suggesting thermally oxidized saturated fatty acids may be precursors of some odors and flavors associated with heated lipids. Presented at the AOCS Fall Meeting, Philadelphia, September 1974. ARS, USDA.     

Emission of Volatile Aldehydes from DAG-Rich and TAG-Rich Oils with Different Degrees of Unsaturation During Deep-Frying

The purpose of the present study was to compare the emission of volatile aldehydes from diacylglycerol-rich oils (DAG-OILs) and triacylglycerol-rich oils (TAG-OILs) with different degrees of unsaturation of fatty acid moieties during the deep-frying of sliced potatoes. To examine the effect of fatty acid composition, four kinds of oils with different fatty acid compositions were selected: rape seed (RS); sunflower oil as a high oleic (HO); safflower oil as high linoleic (HL); and, perilla oil as high linolenic (HLn) oils. The emissions of volatile aldehydes were determined during the deep-frying of sliced potatoes by using the above fresh test oils or deteriorated RS oils. The statistical analysis showed no significant difference in volatile aldehyde emission and profile between the DAG-OIL and TAG-OIL with the fatty acid composition of RS, HL, and HLn. Although a statistically significant difference was noted in the volatile aldehyde emission between the DAG-OIL and TAG-OIL with HO, this difference was extremely small when compared to the variations found in the oils with four types of fatty acid composition. Finally, no difference was found in the volatile aldehyde emissions between the deteriorated DAG-OIL and TAG-OIL, although volatile aldehyde emissions increased with frying time. In addition, the acrylamide contents in potato chips prepared with RS–DAG or RS–TAG were at comparable levels.     

Brain and Liver Headspace Aldehyde Concentration Following Dietary Supplementation with n‐3 Polyunsaturated Fatty Acids

Reactive oxygen species react with unsaturated fatty acids to form a variety of metabolites including aldehydes. Many aldehydes are volatile enough to be detected in headspace gases of blood or cultured cells and in exhaled breath, in particular propanal and hexanal which are derived from omega‐3 and omega‐6 polyunsaturated fatty acids, respectively. Aldehydes are therefore potential non‐invasive biomarkers of oxidative stress and of various diseases in which oxidative stress is thought to play a role including cancer, cardiovascular disease and diabetes. It is unclear, however, how changes in the abundance of the fatty acid precursors, for example by altered dietary intake, affect aldehyde concentrations. We therefore fed male Wistar rats diets supplemented with either palm oil or a combination of palm oil plus an n‐3 fatty acid (alpha‐linolenic, eicosapentaenoic, or docosahexaenoic acids) for 4 weeks. Fatty acid analysis revealed large changes in the abundance of both n‐3 and n‐6 fatty acids in the liver with smaller changes observed in the brain. Despite the altered fatty acid abundance, headspace concentrations of C1–C8 aldehydes, and tissue concentrations of thiobarbituric acid reactive substances, did not differ between the 4 dietary groups. Our data suggest that tissue aldehyde concentrations are independent of fatty acid abundance, and further support their use as volatile biomarkers of oxidative stress.     

Quantitative analysis of free and bound fatty aldehydes: Optimum conditions for p-nitrophenylhydrazone formation

A method is described for the analysis of free and bound fatty aldehydes in lipid extracts. By radiolabeling techniques it was shown that the method for measuring free fatty aldehydes when used in the presence of bound fatty aldehydes measures less than 1% of the bound aldehydes. Parameters of time, temperature and optimum acid concentration are reported. A comparison has been made between the present method and other published methods of measuring p-nitrophenylhydrazones with emphasis on recoveries and reactivities. The present method is suitable for measuring as little as 0.02 μmoles of fatty aldehyde. The method has been applied to the analysis of free and bound aldehydes present in various mouse tissues.     

Deterioration of diacylglycerol‐ and triacylglycerol‐rich oils during frying of potatoes

The stabilities of a commercial diacylglycerol‐rich oil (DAG) and a salad oil (TAG) that had been prepared from a mixture of rapeseed and soybean oils were compared while frying potatoes at 180 °C for 3 h. The representative chemical and physical characteristics of the oils were assessed before and after frying, together with the amount of volatile aldehydes in the exhaust of frying. Among the deterioration indications, the carbonyl value, polymer content, and residual polyunsaturated fatty acid content were similar and not significantly different between the TAG and DAG. On the other hand, the characteristics relating to free fatty acids, i.e. the acid value and emission of chemiluminescence at 100 °C, were greater and the smoke and flash points were lower in the DAG than in the TAG. An irritating odor was generated from the DAG after 1 h of frying and got stronger as frying continued. These results suggested that DAG more easily forms free fatty acids under frying conditions than TAG.     

The autoxidation of highly unsaturated fatty acids: Methyl 4,7,10,13,16,19-docosahexaenoate

Methyl docosahexaenoate was selected as a model system for studying the autoxidation of highly unsaturated fatty acids. The ester was oxidized at 50 C for 28 hr and the volatiles collected by high vacuum distillation and fractionated by gas chromatography. A number of aldehydes, aldehyde esters, hydrocarbons and methyl esters were identified. Mechanisms are proposed for the formation of these compounds.     

Headspace Volatile Oxylipins of Eastern Himalayan Moss Cyathophorella adiantum Extracted by Sample Enrichment Probe

Cyathophorella adiantum (Griff.) M. Fleisch. (Division-Bryophyta, Family-Daltoniaceae), an Eastern Himalayan moss was studied for the first time to identify the volatiles derived from cellular and membrane bound fatty acids. A high capacity sample enrichment probe (SEP) was used for extraction of headspace volatile (HSV) molecules followed by GC–MS analysis. Different short-chain oxylipins like alkenes, alkanes, saturated and unsaturated alcohols, saturated and unsaturated aldehydes, ketones were identified along with free and esterified fatty acids, cyclo compounds and some by-products of secondary metabolites. Fatty acid analysis of neutral lipids (NL) and phospholipids (PL) of this plant exhibits the predominance of C16 and C18 fatty acids. It also reveals some interesting information that might indicate the possible fatty acid precursors for volatile generation and their sources in this plant.     

Volatile hydrocarbons from photosynthetic membranes containing different fatty acids

A cell-free plant system was developed generating short-chain volatile hydrocarbons as markers of light-induced copper-mediated peroxidation of fatty acids present either as endogenous constituents of photosynthetic membranes or added exogenously. Different polyunsaturated fatty acids are present in the blue green algaeAnacystis nidulans, Anabaena variabilis andSpirulina platensis. The first species has no polyunsaturated acids. Thylakoids isolated from these algae produce different short-chain volatile hydrocarbons. The location of the double bond of dienoic or higher polyunsaturated fatty acids most distant from the carboxyl group determines the chain length of hydrocarbons evolved. Their number of C-atoms is the same as found beyond this double bond of the fatty-acid molecule (ω-1). This pattern of volatile hydrocarbons produced is in contrast to thermolytic cleavage. Malondialdehyde is formed only when at least 3 double bonds are present in the fatty acid. Peroxidation of endogenous thylakoidal and added fatty acids is completed within 24 hr; a maximum of 1% of the carbon skeleton can be recovered as volatile hydrocarbons.     

Determination of the amounts of free arachidonic acid in resident and activated rabbit alveolar macrophages by fluorometric high performance liquid chromatography

An improved method of high performance liquid chromatography (HPLC) has been developed for the separation and quantitation of low levels of free fatty acids as they occur in mammalian tissues. The fatty acid analysis is based on the esterification of the carboxylic group with 9-anthryldiazomethane (ADAM). HPLC separation and fluorescence measurement of fatty acid ADAM esters allow the determination of pmole amounts of fatty acids. The amounts of free fatty acids of resident and activated alveolar macrophages were determined by the fluorometric HPLC method. There were approximately 2 μg/10⁶ cells of free fatty acids. In resident macrophages, free 20∶4 was a minor component (0.8% of total free fatty acids), while significant amounts of 20∶4 were found in the total glycerophospholipids, representing 16.6% of the total fatty acids. A marked increase in amounts of 20∶4 (8 times) occurred in activated macrophages stimulated for 1 hr with opsonized zymosan. Small but significant increases (1.5 times) also occurred in other fatty acids. These results show that the release reaction of fatty acids was not selective for 20∶4 in alveolar macrophages after the challenge with opsonized zymosan.     

Lipid oxidation products of heated soybeans as a possible cause of protection from ruminal biohydrogenation

Heating oilseeds has been shown to improve the milk fatty acid profile when given to dairy cows, compared to raw oilseeds. However, results from published studies are conflicting. The conditions of heating and storage of the oilseeds could be responsible for these differences, probably partly through their effects on lipid oxidation, the products of which could act on ruminal biohydrogenation (BH). Thus, 15 different treatments were applied to ground soybeans: three levels of heating (no heating, 30 min at 110 or 150°C) × 5 ambient storage durations (0, 1, 2, 4, or 6 months). Soybeans were incubated in vitro with ruminal fluid for 6 h. Triacylglycerol (TAG) polymers, hydroperoxides and hydroxyacids (HOA), aldehydes, and fatty acids were assayed in soybeans and ruminal culture. No TAG polymer was detected in any treatment. Soybeans stored for a long time had a high content of HOA, whereas those heated at 150°C, whatever the storage duration, had high aldehyde contents. The percentage disappearance of cis‐9,cis‐12 18:2 and cis‐9,cis‐12,cis‐15 18:3 in incubates decreased significantly in cultures with heated soybeans, especially at 150°C, suggesting that this partial protection of polyunsaturated fatty acids (PUFA) from BH was at least in part linked to the aldehyde content of the heated soybeans. Practical applications: Oilseeds given to ruminants are often heated, and heat treatment is known to generate oxidation products. Knowing what oxidation products influence ruminal biohydrogenation of unsaturated fatty acids could result in technological processes allowing a better transfer of unsaturated fatty acids from oilseeds to ruminant products.     

Determination of structure of unsaturated fatty acids via reductive ozonolysis

An improved method of reductive ozonolysis for the determination of structure of unsaturated fatty acids is reported. The ozonization is carried out at −60–−70C by adding the sample dissolved in pentane to a .02–.03 M pentane solution of ozone. The reduction is effected by the Lindlar catalyst at 0C in pentane or in other solvents, such as the methyl esters of short-chain fatty acids or in dimethyl phthalate, and the aldehydic fragments are analyzed by gas-liquid chromatography (GLC). The method is applied to the structural analyses of methyl esters of oleic, linoleic, linolenic, and arachidonic acids. The sensitivity of the method is demonstrated by the analysis of methyl oleate containing a small amount of added methyl linolenate. Mixtures of methyl oleate and petroselinate are analyzed to demonstrate the identification of the simple aldehydes and ester-aldehydes, and to show the potential of the method for quantitative analysis of mixtures of unsaturated esters. Supported in part by the U.S. Public Health Service, N.I.H. Grant H-5735, and in part by a contract with the U. S. Department of Interior, Fish and Wildlife Service. Presented at the AOCS meeting in Chicago, Ill., 1961.     

Deacidification of olive oils by supercritical carbon dioxide

An investigation of the application of supercritical carbon dioxide (SC-CO₂) extraction to the deacidification of olive oils has been made to verify that the nutritional properties of the oil remain unchanged when this technique is applied. Preliminary runs at 20 and 30 MPa in the temperature range of 35–60°C were performed on fatty acids and triglycerides as pure compounds or mixtures, to determine their solubility in SC-CO₂. The solubility data obtained show that CO₂ extracts fatty acids more selectively than triglycerides under specific conditions of temperature and pressure (60°C and 20 MPa). It has been noted that the physical state of the solutes plays an important role in determining the solubility trends as a function of temperature and pressure. Extraction of free fatty acids from olive oil was performed on samples with different free fatty acid (FFA) contents at 20 and 30 MPa and at 40 and 60°C. Experimental data suggest that the selectivity factor for fatty acids is higher than 5 and increases significantly as the fatty acid concentration of the oil decreases. For a FFA content of 2.62%, the selectivity reaches a value of 16. In order to evaluate any variations in the composition, several SC-CO₂ extractions of husk oil with high FFA content (29.3%) were made. The results show that selectivity is still significant (≈5) and the composition in the minor component of the deacidified oil has not changed. On the basis of the experimental results and preliminary process evaluations, the authors conclude that SC-CO₂ extraction could be a suitable technique for the deacidification of olive oils, especially for oils with relatively high FFA (<10%).     

An Improved Method for Determining Medium- and Long-Chain FAMEs Using Gas Chromatography

The existing protocols for analyzing fatty acid methyl esters (FAMEs) using a one-step acetyl chloride (AC) catalyzed transesterification and extraction procedure cannot accurately determine the medium- and long-chain fatty acids simultaneously in clinical (enteral, parenteral) formulations. For example: (1) addition of AC at room temperature generates an exothermic reaction that often results in loss of sample and possible injury to the analyst; (2) certain polyunsaturated fatty acids (PUFAs) are less stable at elevated temperatures during the transesterification and contribute to the over-estimation of the C16:0 and C18:1 fatty acids; and (3) the flame-ionization detector (FID) response varies depending on the carbon chain length of the fatty acids, that consequently impacts the underestimation of medium-chain fatty acid (C6–C10) recoveries. To overcome these deficiencies and accurately determine FAMEs, we have developed an improved one-step transesterification method that employs the addition of AC in tubes kept on a dry ice bath, the transesterification at room temperature, and the data analysis using relative response factors. Using this modified protocol, we determined the fatty acid composition of lipid emulsions (Omegaven^® and Lipidem^®) on a Shimadzu GC2010 gas chromatography (GC) system using a capillary GC column (Zebron ZB-WAX plus, 30 m, 0.25 mm ID, 0.25 μm). Our data suggest that the improved method can be easily used to accurately determine fatty acids (C6–C24) in functional foods and lipid emulsions.     

Industrial chemical uses of polyunsaturated fatty acids

Production of vegetable, animal and marine oils containing more than about 40% unsaturated fatty acids totaled 15,000 million pounds in 1968, almost on the scale of petrochemical production. The greater share (64%) of this nonfossil oil production was directed toward food uses, the remainder toward industrial and animal feed uses. The variety of chemical reactions carried out on these unsaturated fatty acid products include hydrogenation, interesterification, dimerization, sulfation, formation of nitrogen compounds, epoxidation, alkaline cleavage and oxidative ozonolysis. Some of these reactions have been developed at Utilization Research and Development Divisions of the Agricultural Research Service, U.S. Department of Agriculture. Research is continuing in developing new reactions for potential industrial application. An example is reductive ozonolysis of unsaturated fatty esters to produce monofunctional aldehydes and bifunctional aldehyde esters. Presented at the ISF-AOCS World Congress, Chicago, September 1970. No. Market. Nutr. Res. Div., ARS, USDA.     

Volatile compounds in oils after deep frying or stir frying and subsequent storage

Soybean oil (900 g) was heated by deep frying at 200°C for 1 h with the addition of 0, 50, 100, 150 and 200 mL water, and then stored at 55°C for 26 weeks. Soybean oil, corn oil and lard were heated by stir frying and then stored at 55°C for 30 weeks. The volatiles and peroxide values of these samples were monitored. All samples contained aldehydes as major volatiles. During heating and storage, total volatiles increased 260-1100-fold. However, aldehyde content decreased from 62–87% to 47–67%, while volatile acid content increased from 1–6% to 12–33%; especially hexanoic acid which increased to 26–350 ppm in the oils after the storage period was completed. Water addition to the oils heated by deep frying tended to retard the formation of volatile compounds. The total amount of volatile constituents of lard heated by stir frying increased more during storage than that of corn oil or soybean oil. Peroxide values did not reflect the changes of volatile content in the samples.     

Volatile and Semivolatile Compounds in Gray Catbird Uropygial Secretions Vary with Age and Between Breeding and Wintering Grounds

The uropygial secretions of some bird species contain volatile and semivolatile compounds that are hypothesized to serve as chemical signals. The abundance of secretion components varies with age and season, although these effects have not been investigated in many species. We used solid-phase microextraction headspace sampling and solvent extraction coupled with gas chromatography–mass spectrometry to detect and identify volatile and semivolatile chemical compounds in uropygial secretions of gray catbirds (Dumetella carolinensis). We identified linear and branched saturated carboxylic acids from acetic (C2) through hexacosanoic (C26); linear alcohols from decanol (C10) through docosanol (C22); one aromatic aldehyde; one monounsaturated carboxylic acid; two methyl ketones; and a C28 ester. We tested for the effect of age on signal strength and found that juvenile birds produced greater amounts of volatile C4 through C7 acids and semivolatile C20 through C26 acids, although the variation among individuals was large. Adult birds displayed small concentrations and minimal individual variation among volatile compounds, but produced significantly higher levels of long-chain linear alcohols than juvenile birds. We tested for the effects of season/location by sampling adult catbirds at their Ohio breeding grounds and at their Florida wintering grounds and found that the heaviest carboxylic acids are significantly more abundant in secretions from birds sampled during winter at the Florida site, whereas methyl ketones are more abundant in birds sampled during summer on the Ohio breeding grounds. We observed no effect of sex on semivolatile compounds, but we found a significant effect of sex on levels of carboxylic acids (C4 through C7) for juvenile birds only.     

Formation of triacylglycerol core aldehydes during rapid oxidation of corn and sunflower oils with tert-butyl hydroperoxide/Fe2+

The lipid ester core aldehydes formed during a rapid oxidation (7.8 M tert-butyl hydroperoxide, 90 min at 37°C) of the triacylglycerols of purified corn and sunflower oils were isolated as dinitrophenylhydrazones by preparative thin-layer chromatography and identified by reversed-phase high-performance liquid chromatography with on-line electrospray ionization mass spectrometry and by reference to standards. A total of 113 species of triacylglycerol core aldehydes were specifically identified, accounting for 32–53% of the 2,4-dinitrophenylhydrazine (DNPH)-reactive material of high molecular weight representing 25–33% of the total oxidation products. The major core aldehyde species (50–60% of total triacylglycerol core aldehydes) were the mono(9-oxo)nonanoyl- and mono(12-oxo)-9,10-epoxy dodecenoyl- or (12-oxo)-9-hydroxy-10,11-dodecenoyl-diacylglycerols. A significant proportion of the total (9-oxo)nonanoyl and epoxidized (12-oxo)-9,10-dodecenoyl core aldehydes was found in complex combinations with hydroperoxy or hydroxy fatty acyl groups (6–10% of total triacylglycerol core aldehydes). Identified were also di(9-oxo)nonanoylmonoacylglycerols (0.5% of total) and tri(9-oxo)nonanoylglycerols (trace). The identification of the oxoacylglycerols was consistent with the products anticipated from tert-butyl hydroperoxide oxidation of the major species of corn and sunflower oil triacylglycerols (mainly linoleoyl esters). However, the anticipated (13-oxo)-9,11-tridecadienoyl aldehyde-containing acylglycerols were absent because of further oxidation of the dienoic core aldehyde. A significant proportion of the unsaturated triacylglycerol core aldehydes contained tert-butyl groups linked to the unsaturated fatty chains via peroxide bridges (2–9%). The study demonstrates that rapid peroxidation with tert-butyl hydroperoxide consitutes an effective method for enriching natural oils and fats in triacylglycerol core aldehydes for biochemical and metabolic testing.     

Mild Method for Preparation of 4,4-Dimethyloxazoline Derivatives of Polyunsaturated Fatty Acids for GC–MS

A mild and convenient method has been developed for preparing 4,4-dimethyloxazoline (DMOX) derivatives of fatty acids for GC–MS analysis. First, fatty acid methyl esters are converted to corresponding amides by incubation overnight at room temperature with 2-amino-2-methyl-1-propanol and a catalytic amount of sodium methoxide. The resulting 2-(methylpropanol) amides were isolated by partition between hexane–diethyl ether and water, and then converted to 4,4-dimethyloxazoline derivatives by treatment with trifluoroacetic anhydride under mild conditions (50 °C for 45 min). Structures of 2-methylpropanol amide and a DMOX derivative of oleic acid were confirmed by GC–MS. This method was applied to different FAME prepared from animal, plant or microbial lipids. The suggested method is most suitable for structure analysis of polyunsaturated fatty acids (PUFA) and for acids with double bonds in close to terminal positions. Application of the method is illustrated with spectra of the DMOX derivatives of 16:1(n-13), 24:5(n-6) and 24:6(n-3) acids.     

Oxidative stability of natural and randomized high-palmitic-and high-stearic-acid oils from genetically modified soybean varieties

The oxidative stability of soybean oil triacylglycerols (TAG) obtained from genetically modified soybeans was determined before and after chemical randomization. Soybean oil oxidative studies were carried out under static oxygen headspace at 60°C in the dark and oxidative deterioration was monitored by peroxide value, monometric and oligomeric oxidation products, and volatile compounds. Randomization of the soybean oil TAG improved the oxidative stability compared to the natural soybean oil TAG. Oxidative stability was improved by three factors. Factor one was the genetic modification of the fatty acid composition in which polyunsaturated acids (such as linolenic and linoleic acids) were decreased and in which monounsaturated fatty acids (such as oleic) and saturated acids (palmitic and stearic) were increased. Factor two was the TAG compositional modification with a decrease in linolenic and linoleic-containing TAG and an increase in TAG with stearic and palmitic acids in combination with oleic acid. Factor three was the TAG structure modification accomplished by an increase in saturated fatty acids and a decrease in linoleic and linolenic acids at the glycerol moiety carbon 2. Presented at the AOCS Annual Meeting & Expo, Chicago, IL, May 10–13, 1998.     

Effects of some leaf-emitted volatile compounds on aphid population increase

A role of some volatile compounds produced by plant tissues may be as defensive molecules against various pests, including arthropods. Volatile six-carbon compounds derived in plant tissue from polyunsaturated fatty acids via lipoxygenase/hydroperoxide lyase reduced tobacco aphid fecundity at certain concentrations when added to headspace vapor to which aphids were exposed. Both C₆ aldehydes and alcohols were effective, with the alcohols having greater activity. (Z)-3-Hexenyl acetate at levels in the headspace similar to those of the alcohols and aldehydes did not reduce aphid fecundity. A 6-hr exposure period to the C₆ aldehydes and alcohols was needed for maximum effect on the aphids feeding on tobacco leaves. Analysis of the direct versus indirect effects of these compounds indicates that the volatile aldehydes had both direct effects on aphid fecundity and indirect effects due to induced changes in the leaves upon which the aphids were feeding, while only indirect effects were observed for the alcohols. Tomato leaves have the capacity to produce volatile compounds at levels that impact aphid population increase, with the volatiles produced from crushed leaves having a much larger effect. The C₆ aldehydes and alcohols may be components of the fecundity reduction seen with tomato volatiles; however, volatile terpenes showed no effect. These results can be of significance for the genetic alteration of plants for improved aphid resistance.The investigation reported in this paper (No. 93-3-1) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with approval of the Director.