Gas chromatographic separation of the fatty acid methyl esters prepared from dairy products,partially hydrogenated oils,and refined vegetable oils applying a negative temperature gradient

To date no single gas chromatographic method can simultaneously measure all fatty acids (FA), including trans-FA (TFA), that are contained in dairy products, partially hydrogenated oils (PHO), and refined vegetable oils. Using 100% poly(biscyanopropyl siloxane) capillary columns, ruminant and dairy fats are preferentially analyzed by applying temperature programs that separate short chain FA, but not trans-18:3 from 20:1. Refined vegetable oils and PHO are preferentially analyzed by applying isothermal elutions that provide quantification of all 18 carbon TFA including trans-18:3 FA, but not of all short chain FA. In this short communication, we propose a temperature program method capable of simultaneously measuring short chain FA and all 18 carbon TFA including trans-18:3 by applying a negative temperature gradient after the elution of trans-18:1. A simplified version of the method is also described for equipment not able to perform negative temperature gradients.     

A multivariate study of the relationship between fatty acids and volatile flavor components in olive and walnut oils

The relationships between FA and the volatile profiles of olive and walnut oils from Argentina were studied using GC and solid-phase microextraction coupled with GC-MS. The major volatiles were aldehydes and hydrocarbons, produced mainly through the oxidative pathways. n-Pentane, nonanal, and 2,4-decadienal were predominant in walnut oils, whereas nonanal, 2-decenal, and 2-undecenal were the most abundant components in olive oils. A multivariate analysis applied to the chemical data emphasized the differences between the oils and allowed us to see a pattern of covariation among the FA and the volatile compounds. The main differences between walnut and olive oils were the presence of larger amounts of short-chain (C₅–C₆) saturated hydrocarbons and aldehydes in the former and the greater quantities of medium-chain (C₇–C₁₁) compounds in olive oil. This can be explained by their different origins, mainly from the linoleic acid in walnut oil or almost exclusively from the oleic acid in olive oil.     

Selective enzymatic synthesis of lower acylglycerols rich in polyunsaturated fatty acids

Selective elimination of saturated fatty acid (FA) residues from vegetable oils (sesame, borage, olive, and sunflower) produces lower acylglycerols rich in unsaturated and polyunsaturated FA residues. Novozym® 435 releases greater amounts of saturated FA residues than unsaturated residues relative to other biocatalysts. Highest selectivity is obtained for 50 wt‐% Novozym® 435 with respect to the oil and [ethanol]/[FA] = 25. Under these conditions, 88–90% of saturated residues are released versus only 44–45% of unsaturated residues. Product acylglycerols have ratios of unsaturated to saturated residues of 21.5–23.0 relative to 5.22–4.73 for the precursor oils. Ethanolysis is faster with Novozym® 435. This biocatalyst is much less susceptible to inactivation by ethanol than other biocatalysts. However, in excess ethanol ([EtOH]/[FA] = 4.5–25), the rate and conversion both increased when the ethanol concentration decreased. The best combination of conversion and selectivity is obtained with 50 wt‐% Novozym® 435 and [ethanol]/[FA] = 4.5. Under these conditions, the product obtained in 1 h contains ca. 48% triacylglycerols, 24% diacylglycerols and 28% monoacylglycerols; the mol percentage of unsaturated FA residues is 12.3 times greater than that of saturated residues. The sn‐2 position is essentially free of saturated FA residues. After use for ten reaction cycles, Novozym® 435 retains 80% of its initial activity.     

Selective Seasonal Fatty Acid Accumulation and Mobilization in the Wild Raccoon Dog (<Emphasis Type="Italic">Nyctereutes procyonoides</Emphasis>)

Previous studies on laboratory rodents, rabbits and humans have demonstrated that fatty acid (FA) mobilization from white adipose tissue (WAT) is selective and its efficiency is related to FA structure. Selective FA mobilization was also documented in a carnivore, the farmed raccoon dog (Nyctereutes procyonoides), fasted for 8 weeks. The present study explored whether similar selectivity of FA mobilization was manifested in wild mammals experiencing seasonal food scarcity and abundance. Fractional mobilization from and incorporation into WAT of a wide spectrum of FA were studied by gas–liquid chromatography from the subcutaneous WAT of free-ranging raccoon dogs with the same individuals sampled in consecutive seasons. The wintertime FA mobilization was selective and mostly confirmed the patterns of FA release in captivity. Mobilization correlated inversely with the FA chain length but increased with unsaturation and when the first double bond was located closer to the methyl end. 18–20C n-3 polyunsaturated FA (PUFA) and 14–17C monounsaturated FA (MUFA) were preferentially mobilized while 19–24C saturated FA and MUFA were preserved during wintering. The summertime FA incorporation correlated inversely with the chain length and increased with unsaturation and in MUFA and PUFA with double bonds closer to the methyl end. The principles of selective FA mobilization were valid in wild mammals. FA incorporation was also selective and reversed the wintertime losses of the preferably mobilized FA.     

Thermal degradation of FA and catfish and menhaden oils at different refining steps

The thermal degradation (weight loss) of individual FA and of catfish and menhaden oils collected from different refining steps was investigated by thermogravimetric analysis. The heat resistance of FA was partially dependent on chain length and degree of unsaturation. The weight loss of catfish and menhaden oils increased with increased heating temperatures, regardless of the oil refining process. All oil samples (except crude catfish oil) were decomposed after the heating temperature reached 550°C. Based on the thermogravimetric curves, the following thermal stability sequence at different refining steps for both catfish and menhaden oils was proposed: crude > degummed > neutralized > bleached > deodorized oils.     

Determining Ethyl Esters in Fish Oil with Solid Phase Microextraction and GC–MS

The long-chain polyunsaturated fatty acids (PUFA) found in fish oil, specifically eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) play an important part in human health. As a result, fish oil supplements are commonly consumed by people around the world. Supplements in the form of triacylglycerols (TAG) can be sold at a premium price, compared to those in the ethyl ester (EE) forms. Producers of TAG supplements require a simple, rapid method to determine the authenticity of their raw material. Here, we describe a method to quantify EE in fish oil using solid phase microextraction headspace analysis and GCMS. Despite the variation in linear ranges of the calibration curves with volatility of the EE, 30 individual FA were quantified including common saturated FA such as palmitic and stearic acid, as well as longer chain PUFA, such as EPA and DHA. The method was then applied to three commercial fish oils in the TAG form and two of the products were found to contain EE, with one containing EE above 1.5% w/w, indicating that contamination had occurred. With growing consumer interest in fish oil products, the method proposed here will help resolve future issues of authenticity in fish oils.     

Zeolite‐catalyzed isomerization of oleic acid to branched‐chain isomers

Branched‐chain (bc) saturated fatty acids (SFA) have potential as oleochemical intermediates since they have better oxidative stability than linear unsaturated fatty acids (UFA) and have better low‐temperature properties than linear SFA. Previous studies in converting UFA to bc‐FA using clay catalysts have resulted in only modest yields and conversions. Recent reports, however, have suggested that certain zeolites can be effective catalysts for converting UFA to bc‐FA in higher yields and conversions. In this work, we examined the scope and potential of the zeolite‐catalyzed synthesis of bc‐FA starting from readily available monounsaturated linear FA. Our results show that common UFA such as oleic acid can be converted to bc‐isomers using modified Ferrierite zeolite catalysts with high conversions (98%) and high selectivity (85%) and that the zeolite catalysts are reusable for at least three cycles. The positions of branching (methyl) on the FA chain were determined from the GC‐MS spectra of the picolinyl esters of the bc‐FA.     

Elongation of very Long-Chain (>C<Subscript>24</Subscript>) Fatty Acids in <Emphasis Type="Italic">Clarias gariepinus</Emphasis>: Cloning,Functional Characterization and Tissue Expression of <Emphasis Type="Italic">elovl4</Emphasis> Elongases

Elongation of very long‐chain fatty acid 4 (Elovl4) proteins participate in the biosynthesis of very long‐chain (>C₂₄) saturated and polyunsaturated fatty acids (FA). Previous studies have shown that fish possess two different forms of Elovl4, termed Elovl4a and Elovl4b. The present study aimed to characterize both molecularly and functionally two elovl4 cDNA from the African catfish Clarias gariepinus. The results confirmed that C. gariepinus possessed two elovl4‐like elongases with high homology to two previously characterized Elovl4 from Danio rerio, and thus they were termed accordingly as Elovl4a and Elovl4b. The C. gariepinus Elovl4a and Elovl4b have open reading frames (ORF) of 945 and 915 base pairs, respectively, encoding putative proteins of 314 and 304 amino acids, respectively. Functional characterization in yeast showed both Elovl4 enzymes have activity towards all the PUFA substrates assayed (18:4n‐3, 18:3n‐6, 20:5n‐3, 20:4n‐6, 22:5n‐3, 22:4n‐6 and 22:6n‐3), producing elongated products of up to C₃₆. Moreover, the C. gariepinus Elovl4a and Elovl4b were able to elongate very long‐chain saturated FA (VLC‐SFA) as denoted by increased levels of 28:0 and longer FA in yeast transformed with elovl4 ORF compared to control yeast. These results confirmed that C. gariepinus Elovl4 play important roles in the biosynthesis of very long‐chain FA. Tissue distribution analysis of elovl4 mRNAs showed both genes were widely expressed in all tissues analyzed, with high expression of elovl4a in pituitary and brain, whereas female gonad and pituitary had the highest expression levels for elovl4b.     

FA composition of the oil extracted from farmed atlantic salmon (Salmo salar L.) viscera

The FA composition of visceral oil extracted from farmed Atlantic salmon (Salmo salar L.) viscera was studied. Seventeen FA were identified in the extracted visceral oil, and the major FA were 18∶1n9, 16∶0, 16∶1n7, 20∶5n3 (EPA), 14∶0, and 22∶6n3 (DHA). The percentages of saturated, monounsaturated, and polyunsaturated FA in the total FA were 31.7, 36.0, and 32.2%, respectively. Compared with other fish oils, oil from farmed Atlantic salmon had much higher EPA (1.64 g/100 g) and DHA (1.47 g/100 g) contents. The FA profile of the salmon visceral oil was similar to that of the salmon fillet. Thus, the salmon visceral oil could be a replacement for the oil obtained from edible salmon fillet and used in functional foods or feeds requiring a high level of omega-3 FA. Furthermore, producing visceral oil is also beneficial to salmon fish industry by adding value back to the processing waste.     

Correlating chemical structure and physical properties of vegetable oil esters

The influence of FA ester chemical structures on the rheology and crystallization temperature of those compounds was evaluated using methyl, n-butyl, n-octyl, and 2-ethyl-1-hexyl FA esters with different chain lengths and different degrees of unsaturation. The rheological properties were analyzed in a high-precision rheometer at various temperatures, and the crystallization temperatures were determined by DSC. Esters produced from the esterification of pure FA and from the transesterification of vegetable oils (i.e., soybean, corn, linseed, and babassu coconut oils) were evaluated. The length of the FA chain was shown to have a marked influence on the viscosity and crystallization temperature of the systems, whereas branching affected only the crystallization temperature to a significant extent. The viscosity and crystallization temperature of the systems were also influenced by the degree of unsaturation. One double bond was shown to increase viscosity, whereas two or three double bonds caused a decrease in the viscosity of the systems. Unsaturation lowered the crystallization temperature in all cases, regardless of the number of double bonds. From all the oils studied, methyl esters from babassu coconut oil presented the lowest crystallization temperatures.     

Fatty acid profiles of 80 vegetable oils with regard to their nutritional potential

The current concern for fat intake in western countries has raised the question of the individual fatty acid (FA) impact on health. This important issue has strengthened the awareness of nutritionists and food manufacturers for the control of the FA profile of food products. The aim of this review is to provide a classification of the FA profiles of 80 vegetable oil sources, according to their nutritional potential. The first part of the review focuses on lipoprotein metabolism, and on the impact of each dietary FA on blood lipid composition (LDL‐cholesterol, HDL‐cholesterol and circulating triacylglycerols). In the second part of the review, the oil sources are clustered by similar FA profiles, and the classification is discussed with regard to the individual FA action on blood lipid composition. Apart from the major vegetable seeds, the clustering highlighted some interesting nutritional oil sources containing mainly α‐linolenic acid (camelina, linseed, perilla and stock oils), or interesting amounts of the two essential FA (purslane, chia, raspberry seed, sea buckthorn seed and salicorn oils). Furthermore, this classification provides a useful tool for the formulation of the FA profile of food products.     

Structuring of edible oils by long-chain FA,fatty alcohols,and their mixtures

Homologous series of fatty alcohols and FA with chain lengths ranging from 16 to 22 carbons were evaluated for their oil-structuring potential by texture profile analysis (TPA). FA, fatty alcohols, and their mixtures were found to structure sunflower oil and several vegetable oils at levels as low as 2% (w/w) for the pure ingredients. Mixtures of fatty alcohols and FA with the same chain lengths, at 5% (w/w) in sunflower oil, showed a synergistic effect below 20°C at the composition ratios of 7∶3 and 3∶7 (w/w). This synergistic effect in the 7∶3 stearyl alcohol/stearic acid mixture was due to effects on the microstructure of the composite material. The larger number of small crystals observed in the mixture was attributed to effects on the crystallization kinetics as a result of minimal interfacial tension at the specific 1∶3 and 3∶1 molecular ratios.     

Characterization of cold-pressed onion, parsley, cardamom, mullein, roasted pumpkin, and milk thistle seed oils

Cold-pressed onion, parsley, cardamom, mullein, roasted pumpkin, and milk thistle seed oils were characterized for their fatty acid (FA) composition, tocopherol content, carotenoid profile, total phenolic content (TPC), oxidative stability index (OSI), color, physical properties, and radical-scavenging capacities against peroxyl (oxygen radical-scavenging capacity) and stable DPPH (diphenylpicrylhydrazyl) radicals. Parsley seed oil had the highest oleic acid content, 81 g/100 g total FA, and the lowest saturated fat among the tested oils. Roasted pumpkin seed oil contained the highest level of total carotenoids, zeaxanthin, β-carotene, cryptoxanthin, and lutein at 71 μmol/kg and 28.5, 6.0, 4.9, and 0.3 mg/kg oil, respectively. Onion seed oil exhibited the highest levels of α- and total tocopherols under the experimental conditions. One of the parsley seed oils exhibited the strongest DPPH scavenging capacity and the highest oxygen radical absorbance capacity (ORAC) value of 1098 μmol Trolox equiv/g oil. However, ORAC values of the tested seed oils were not necessarily correlated to their DPPH scavenging capacities under the experimental conditions. The highest TPC of 3.4 mg gallic acid equiv/g oil was detected in one of the onion seed oils. The OSI values were 13.3, 16.9–31.4, 47.8, and 61.7 h for the milk thistle, onion, mullein, and roasted pumpkin seed oils, respectively. These data suggest that these seed oils may serve as dietary sources of special FA, tocopherols, carotenoids, phenolic compounds, and natural antioxidants. An erratum to this article is available at .     

Wax analysis of vegetable oils using liquid chromatography on a double-adsorbent layer of silica gel and silver nitrate-impregnated silica gel

A chromatographic method is described to measure the crystallizable wax content of crude and refined sunflower oil. It can also be applied to any other vegetable oil. The preparative liquid chromatography step on a glass column containing a silica gel adsorbent superimposed upon a silver nitrate-impregnated silica gel support is used to isolate a wax fraction which is then analyzed by gas chromatography. The recovered wax fraction contains, in addition to the crystallizable waxes, hydrocarbons and other compounds with gas chromatographic retention times corresponding to waxes with chain lengths C₃₄−C₄₂. These compounds are short-chain saturated waxes in fruit oils, such as grapeseed and pomace. In seed oils such as sunflower, soybean or peanut, the compounds initially referred to as “soluble esters” are identified as monounsaturated waxes, esters of long-chain saturated fatty acids, and a monounsaturated alcohol, mainly eicosenoic alcohol. Such waxes are absent from corn or rice bran oils.     

Diet,Plasma, Erythrocytes,and Spermatozoa Fatty Acid Composition Changes in Young Vegan Men

There has been increasing interest in vegan diets, but how this dietary pattern regulates tissue fatty acids (FA), especially in men, is unclear. Our aim was to evaluate the effect of a vegan diet on plasma, erythrocyte, and spermatozoa FA composition in young men. Two groups consisting of 67 young (18–25 years old) men were studied. One group following an omnivore diet but did not consume fish, shellfish or other marine foods (control, n = 33), and another group following a vegan diet (vegan, n = 34) for at least 12 months were compared. Dietary intake was assessed via a food frequency questionnaire and a 24-h recall. FA composition was measured in plasma, erythrocyte phospholipids, and spermatozoa by gas–liquid chromatography. Compared to controls, the vegan group had higher reported intakes of carbohydrate, dietary fiber, vitamins (C, E, K, and folate), and minerals (copper, potassium) but lower intakes of cholesterol, trans FA, vitamins B₆, D, and B₁₂, and minerals (calcium, iron, and zinc). Vegan's reported a lower saturated FA and not arachidonic acid intake, both groups did not intake eicosapentaenoic acid and docosahexaenoic acid (DHA), but vegan's showed a higher alpha linolenic acid ALA intake. Vegans had higher plasma, erythrocyte phospholipid, and spermatozoa ALA, but lower levels of other n-3 polyunsaturated fatty acid (PUFA), especially DHA. Vegans were characterized by higher ALA, but lower levels of other n-3 PUFA, especially DHA in plasma, erythrocytes, and spermatozoids. The biological significance of these findings requires further study.     

Identification and Characterization of Phospholipids with Very Long Chain Fatty Acids in Brewer’s Yeast

Yeast lipids and fatty acids (FA) were analyzed in Saccharomyces pastorianus from seven breweries and in the dietary yeast supplement Pangamin. GC–MS identified more than 30 FA, half of which were very‐long chain fatty acids (VLCFA) with hydrocarbon chain lengths of ≥22 C atoms. Positional isomers ω‐9 and ω‐7 were identified in FA with C18–C28 even‐numbered alkyl chains. The most abundant ω‐7 isomer was cis‐vaccenic acid. The structure of monounsaturated FA was proved by dimethyl disulfide adducts (position of double bonds and cis geometric configuration) and by GC–MS of pyridyl carbinol esters. Ultra‐high performance liquid chromatography‐tandem mass spectrometry with negative electrospray ionization identified the phospholipids phosphatidylethanolamine, phosphatidylinositol and phosphatidylcholine, with more than 150 molecular species. Wild‐type unmutated brewer's yeast strains conventionally used for the manufacture of food supplements were found to contain VLCFA.     

Hydrocarbons and alcohols of basking shark and pig liver lipids

Four samples of the unsaponifiables of basking shark liver oil were adsorbed on alumina and eluted to yield Fractions 1–5, inclusive. Analyses by temperature programmed GC and by silica gel chromatography showed hydrocarbons in the first four fractions with squalene increasing to Fraction 3 and the pristane level being highest in Fraction 1. Aside from pristane and squalene, other hydrocarbons occurred at levels of 420–750 mg% in the oils on a weight basis, of which about 60% constituted a series of n-paraffins (relative carbon number range: 15.0–38.0) together with smaller amounts of at least one branched chain saturated group. Unsaturated hydrocarbons eluted mainly after squalene. The oils contained up to 460 mg% sterol and 78–270 mg% alcohols of C₁₀ to C₃₀, the ratio of saturated to unsaturated members being about 1.6. The composition of the unsaponifiable lipids of pig liver was quite different from that of the marine oils. It contained 10.6% sterol in addition to 400 mg% alcohols, the latter consisting of 81.8% saturated components (C₁₂ to C₃₁; ratio of saturated: unsaturated members, 4.4). The hydrocarbons comprised 450–700 mg% of the unsaponifiable mixture and squalene, paraffins and additional unsaturated components occurred at levels of 20.6, 24.4 and 11.9 mg%, respectively. The saturated hydrocarbons were high in normal homologs of relative carbon number range, 15 to 36; pristane could not be detected.     

Surface activity of the lipid products hydrolyzed with lipase and phospholipase A-2

We examined thein vitro surface activity, immersional wettability and adhesional wettability shown by aqueous solutions of soy lysophospholipid (SLP)/monoglyceride (MG)/fatty acid (FA), SLP/FA and SLP/MG, and found that many lipid mixtures showed significant surface activity when their MG and FA components consisted of polyunsaturated FA and/or medium chain FA. The more unsaturated the FA, the higher the surface activity. A mixture of SLP/medium chain fatty acid MG (medium chain MG)/medium chain FA showed the highest surface activity, and was comparable to an Aerosol-OT surfactant, the most effective wetting agent. SLP/polyunsaturated FA monoglyceride (polyunsaturated MG)/polyunsaturated FA, SLP/medium chain FA, and SLP/polyunsaturated FA, SLP/medium chain MG, and SLP/polyunsaturated MG also showed a high degree of activity. Wettability decreased rapidly when the amount of saturated, long chain FA moieties increased. It is recognized that the degrees of unsaturation and the chain length of FAs in the lipid mixtures have a decisive influence on surface activities. Higher ratios of MG and FA to SLP gave higher activity; and solubilizers such as bile salts were necessary to dissolve them in water.     

Oxidative decarboxylation of unsaturated fatty acids

Long‐chain internal olefins were prepared by silver(II)‐catalyzed oxidative decarboxylation of unsaturated fatty acids by sodium peroxydisulfate. Similar to saturated carboxylic acids, 1‐alkenes were the major decarboxylation product in the additional presence of copper(II), whereas in the absence of copper(II) alkanes were predominantly formed. In both cases, the internal unsaturation of the fatty acids remained largely intact, although the moderate yields indicated that side reactions occurred to a significant extent. The simple procedure makes this multistep one‐pot reaction useful for the synthesis of a variety of internally unsaturated hydrocarbons. The purified products, almost all of which are prepared for the first time, may serve as reference compounds for studies on the heterogeneously catalyzed decarboxylation of triglycerides and fatty acids in the absence of hydrogen. Practical applications: The products of the chemistry described in this contribution, i.e., unsaturated long‐chain hydrocarbons, provide bio‐based building blocks for further chemical modification toward products which may be applied as (bio)fuels, lubricants, solvents, and polymeric materials.     

Photoelectron resonance capture ionization mass spectrometry of fatty acids in olive oil

Photoelectron resonance capture ionization (PERCI) mass spectrometry has been developed for the direct online analysis of organics, including lipids. Analysis is conducted without the need for sample preparation or chemical derivatization such as methylation, foregoing the use of harmful or toxic chemicals. PERCI is currently being adapted towards the analysis of edible oils. Herein, as a proof of principle of the simplicity and potential utility of this method towards the analysis of edible oils, we present the analysis of the prevalent fatty acids (FA) in Tuscan extra‐virgin olive oil along with triolein and linolenic acid (LNA) standards. Ionization of olive oil results in little fragmentation of the prevalent FA, which are measured as their molecular ions, [FA–H]^?. The relative concentrations of these FA determined by PERCI were in good agreement with established values. Further utility of PERCI was demonstrated by interrogation of ozonized LNA and olive oil, with 13 of the 18 anticipated products of the ozonolysis of LNA measured as their molecular anions, [M–H]^?. Similarly, the PERCI mass spectrum of ozonized olive oil showed all the anticipated ions of the predominant FA, oleic acid, as well as many molecular ions arising from less abundant unsaturated FA.