Direct quantification of fatty acids in dairy powders with special emphasis on trans fatty acid content

In this study a validated procedure for accurate determination of fatty acids in dairy products, with special emphasis on total trans fatty acids (TFA) content is presented. Dairy fat naturally contains 4–6% of trans fatty acids, mainly trans-octadecenoic acids (i.e. vaccenic acid), and 0.3–1.5% of conjugated linoleic acids (CLA). The proposed procedure does not require lipid extraction, and transesterification of lipids could be carried out directly on dairy products. Optimal analytical conditions have been developed to allow accurate determination of TFA content without prior fractionation of cis/trans FAME isomers by thin-layer chromatography. The methodology requires the use of a highly polar open tubular capillary column having at least 100 m length. CLA and other fatty acids from C4:0 (butyric) acid to long-chain polyunsaturated fatty acids (LC-PUFAs) could also be analyzed. Therefore, the methodology presented is versatile and could be used for both targeted analysis (e.g. determination of TFA in dairy products) and determination of the broad fatty acid profile in dairy products.     

Formation of trans fatty acids in edible oils during the frying and heating process

To assess an impact of heated edible oils on intake of trans fat, the formations of trans fatty acids (TFAs) in cooking conditions was estimated by a frying and heating model system. For the frying model, sliced raw potatoes (10% of the frying oil (w/w)) were fried in commercially available canola oil at 160, 180 and 200 °C, and the 10 frying cycles were performed. The TFAs contained both in fried potatoes and in frying oils were measured by gas chromatography (GC). Lipids content of raw potatoes was about 0.1% (w/w) and TFAs in the raw potatoes were negligible. On the other hand, fried potatoes contained lipids at the level of 8.8%–9.2% and their fatty acid composition was mostly in correspondence with that of the frying oil. The TFAs amount of potatoes fried by the tenth frying operation was at the level of 0.99–1.05 g/100 g lipids. When 100 g potatoes fried in this process were consumed, the TFAs intake was estimated at less than 0.1 g. After 10 frying operations, TFAs content, acid values and peroxide values of the frying oils were measured and compared with those of corresponding heated canola oils without food. The amounts of trans 18:1 FAs contained both in the frying oil and in heated oil were less than the quantitative limit (0.047 g/100 g oil). The increases of trans 18:2 FAs and trans 18:3 FAs of the used frying oil were 0.02 g/100 and 0.05 g/100 g, respectively, compared with those of the fresh oil. trans 18:2 FAs accumulation in the heated oil was slightly less than that in the frying oil. To elucidate TFAs accumulation in various edible oils during cooking, six kinds of commercially available edible vegetable oils were heated to 180 °C in glass test tubes. Small changes in TFAs amounts were observed after four hours heating. These results suggested that an ordinary frying process using unhydrogenated edible oils has little impact on TFAs intake from edible oils.     

Effects of antioxidants on heat-induced trans fatty acid formation in triolein and trilinolein

To elucidate the relation between heat-induced cis/trans isomerisation and thermal oxidative degradation of double bonds in unsaturated lipids, we investigated the effects of several edible antioxidants on these two molecular structural changes of double bonds in triolein (cis-9, 18:1) and trilinolein (cis-9, cis-12, 18:2), which were heated at 180 °C. trans Isomerisation and oxidative degradation of each cis double bond in the triacylglycerols during heating were evaluated on the basis of the increase in the amount of trans isomers and decrease in the amount of cis isomers by gas chromatography analysis. The synchronous suppression in trans isomerisation and cis deterioration of double bonds in these triacylglycerols were found by addition of antioxidants, whose inhibitory effects were associated with their kinds and concentrations. When triolein was heated in the presence of antioxidant, suppression of heat-induced trans isomerisation was directly proportional to that of cis deterioration. The results of our analysis suggested that the appropriate addition of antioxidants to edible oils during processing and cooking would facilitate the control of not only thermal oxidative degradation but also heat-induced trans isomerisation of double bonds in unsaturated lipids.     

Effect of weaning status on lipids of Galician Blond veal: Total fatty acids and 18:1 cis and trans isomers

The effect of weaning at different ages (NW = not weaned, W5 = 5.5 months and W2 = 2 months) on fatty acids (FA) of the Longissimus thoracis (LT) muscle was studied in 36 Galician Blond (GB) calves. Total FA (TFA) were determined by gas chromatography (GC) and 18:1 isomers by a combination of reversed-phase high performance liquid chromatography (HPLC) and GC. NW group showed higher (P < 0.001) values of n-3 polyunsaturated FA (PUFA), conjugated linoleic acid (CLA) and 18:1trans-11 compared to LT from W5 and W2 calves. W2 calves showed the highest levels of n-6 PUFA (P < 0.01), 18:1trans-10 and 18:1trans-6/7/8 (P < 0.001). Generally, W5 calves had intermediate values for TFA and 18:1 isomers. As the suckling period was longer, GB milk and veal FA profiles became more similar, it seems that muscle FA were partially transmitted by the milk FA intake due to the persistence of the reticular grove closing reflex.     

Analysis and formation of trans fatty acids in hydrogenated soybean oil during heating

Hydrogenated oil has been widely used for production of shortenings or margarine, however, the presence of trans fatty acids may be detrimental to human health. The objectives of this study were to develop an improved method for analysis of trans fatty acids and evaluate their formation in both unhydrogenated and hydrogenated soybean oil during heating at 160, 180 and 200 °C for varied length of time. Results showed that among the four columns tested, an Agilent HP-88 column (100 × 0.25 mm I.D., 0.2-μm film thickness) could resolve eight trans fatty acids and nine cis fatty acids simultaneously within 31 min with injector temperature 240 °C, detector temperature 250 °C, and column temperature 170 °C in the beginning, maintained for 24 min, increased to 220 °C at 7.5 °C/min, 230 °C at 10 °C/min, and maintained for 5 min. The contents of both cis and trans fatty acids showed a decreased trend for the increase of heating time or temperature. No trans fatty acid formation was observed even after extensive heating of unhydrogenated and hydrogenated soybean oil for 24 h. This phenomenon demonstrated that trans fatty acids can only be formed under severe conditions.     

Effect of Ganoderma lucidum (Reishi mushroom) or Olea europaea (olive) leaves on oxidative stability of rabbit meat fortified with n-3 fatty acids

The objective of the present study was to evaluate the effect of Ganoderma lucidum (Reishi mushroom) or Olea europaea (olive tree) leaves on oxidative stability of rabbit meat fortified with n-3 fatty acids. Forty-eight slovenska kunka (SIKA) rabbits were divided into four homogeneous groups. The control group ( CONT−) received diet with 6% palm fat; other groups received diet with 6% linseed oil and were either unsupplemented (CONT +) or supplemented with 1% of G. lucidum (REISHI) or O. europaea leaves (OLIVE). Rabbits were slaughtered and fatty acid composition, concentration of vitamin E and malondialdehyde (MDA) in back muscle were analyzed. The results showed that linseed oil addition improved fatty acid composition by increasing polyunsaturated fatty acid (PUFA) proportion, decreasing proportion of saturated fatty acid (SFA) and reducing n-6/n-3 ratio in rabbit meat. Groups that were supplemented with linseed oil had lower content of α-tocopherol and higher content of γ-tocopherol, compared to the CONT − group. The addition of potential antioxidants did not effectively prevent oxidation of rabbit meat.     

伊拉兔肉肌内脂肪酸组成及温度对其脂肪氧化的研究

研究了伊拉肉兔后腿肉的肌内脂肪含量,并分析测定了其脂肪酸组成,同时研究了兔腿肉在常温(15±0.5)℃和低温(4±0.5)℃贮藏条件下的脂肪氧化稳定性。结果表明,兔腿肉肌内脂肪主要含有19种脂肪酸,其中棕榈酸(16∶0)、硬脂酸(18∶0)、油酸(18∶1)、亚油酸(18∶2)的总含量占脂类总脂肪酸含量的71.06%左右。伊拉兔肉肌内脂肪中主要是不饱和脂肪酸,尤以多不饱和脂肪酸居多。在常温(15±0.5)℃和低温(4±0.5)℃贮藏条件下,兔腿肉POV、TBA值的差异极显著(p<0.01)。常温条件下,第5d兔肉出现异味并逐渐变成臭味,而低温条件下,第7d才开始出现轻微异味,低温冷藏对保证肉品质量有重要意义。      

The effect of heat treatment on the cholesterol oxides,cholesterol, total lipid and fatty acid contents of processed meat products

The effects of heat treatment on the formation of cholesterol oxides and on alterations of fatty acid composition were investigated in processed meat products. Meatballs (beef), hamburger (beef and Chester), sausage (pork, chicken and Chester) and frankfurter (mixed meat, chicken and Chester) were analysed. There was no cholesterol oxide formation caused by heat treatment of the samples analysed. The fatty acid compositions, calculated as g/100 g sample, showed alterations only between the raw and grilled beef hamburger. Only the cholesterol levels were significantly changed when comparing the raw and grilled pork sausages and the raw and grilled Chester hamburger, the values being lower in the grilled samples. Also, the total lipid contents of grilled beef hamburgers were lower than the values.     

Effect of different cooking methods on lipid oxidation and formation of free cholesterol oxidation products (COPs) in Latissimus dorsi muscle of Iberian pigs

The aim of this work was to study the influence of different cooking methods (grilled (GR), fried (FP), microwave (MW) and roasted (RO)) on lipid oxidation and formation of free cholesterol oxidation products (COPs) of meat from Iberian pigs that have been fed on an intensive system. Moisture and total lipid content, TBARs, hexanal and COPs were measured in Latissimus dorsi muscle samples. Cooking did not produce changes in total lipid content in meat but induced significantly higher lipid oxidation (TBARs and hexanal values) (p < 0.001) and cholesterol oxidation (COPs) (p < 0.01). When the different cooking methods were studied, the grilled method was the least affected by lipid oxidation (TBARs and hexanal) compared to the others. There were no significant differences among different cooking methods on COPs values. The most abundant cholesterol oxides were both 7α-hydroxycholesterol and 7β-hydroxycholesterol in all groups studied.     

GC-MS quantification of fatty acid profile including trans FA in the locally manufactured margarines of Pakistan

Ten margarine brands of Pakistan were analyzed for their fatty acid composition with emphasis on trans fatty acids (TFA) using GC-MS. Saturated, cis-monounsaturated and polyunsaturated fatty acids were present at 24.2–58.1, 5.7–35.4 and 3.8–37.4% of total fatty acids, respectively. Among the saturated fatty acids, palmitic acid (16.9–33.8%) was dominant in all analyzed margarine brands and its higher amount indicates that palm oil was a major contributor in the margarine manufacturing. Among samples tested only one contained a low level of TFA (2.2%) while the rest contained very high amounts of TFA (11.5–34.8%) which clearly shows that hydrogenated oils were used in the formulation of margarines. Fatty acid profiles demonstrated that all samples belong to the hard margarine category containing high amounts of trans and saturated fatty acids which is an alarming issue for the health of consumers.     

Milk fatty acid profile and dairy sheep performance in response to diet supplementation with sunflower oil plus incremental levels of marine algae

In an attempt to develop strategies for enhancing the nutritional value of sheep milk fat, dairy ewe diet was supplemented with 3 incremental levels of marine algae (MA), in combination with sunflower oil, to evaluate the effects of these marine lipids on milk fatty acid (FA) profile and animal performance. Fifty Assaf ewes in mid lactation were distributed in 10 lots of 5 animals each and allocated to 5 treatments (2 lots per treatment): no lipid supplementation (control) or supplementation with 25 g of sunflower oil/kg of DM plus 0 (SO), 8 (SOMA₁), 16 (SOMA₂), or 24 (SOMA₃) g of MA (56.7% ether extract)/kg of DM. Milk production and composition, including FA profile, were analyzed on d 0, 3, 7, 14, 21, and 28 of treatment. Neither intake nor milk yield were significantly affected by lipid addition, but all MA supplements decreased milk fat content from d 14 onward, reaching a 30% reduction after 28 d on SOMA₃. This milk fat depression might be related not only to the joint action of some putative fat synthesis inhibitors, such as trans-9,cis-11 C18:2 and probably trans-10 C18:1, but also to the limited ability of the mammary gland to maintain a desirable milk fat fluidity, that would have been caused by the noticeable increase in trans-C18:1 together with the lowered availability of stearic acid for oleic acid synthesis through Δ⁹-desaturase. Furthermore, all lipid supplements, and mainly MA, reduced the secretion of de novo FA (C6:0-C14:0) without increasing the yield of preformed FA (>C16). Supplementation with sunflower oil plus MA resulted in larger increases in cis-9,trans-11 C18:2 than those observed with sunflower oil alone, achieving a mean content as high as 3.22% of total FA and representing a more than 7-fold increase compared with the control. Vaccenic acid (trans-11 C18:1) was also significantly enhanced (on average +794% in SOMA treatments), as was C22:6 n-3 (DHA) content, although the transfer efficiency of the latter, from the diets to the milk, was very low (5%). However, the highest levels of MA inclusion (SOMA₂ and SOMA₃) reduced the milk n-6:n-3 ratio, but MA supplements caused an important increase in trans-10 C18:1, which would rule out the possibility that this milk has a healthier fat profile before determining the specific role of each individual FA and ensuring that this trans-FA is at least innocuous in relation to cardiovascular disease risk.     

Muscle lipids and fatty acid profiles of three edible fish from the Mauritanian coast: Epinephelus aeneus, Cephalopholis taeniops and Serranus scriba

Muscle lipids and fatty acids (FA) of three largely consumed seawater species of Serranidae (Epinephelus aeneus, Cephalopholis taeniops, and Serranus scriba) from the Mauritanian coast, were determined through 1 year. The lipid contents were relatively poor, ranging from 0.8% to 2.3% showing a significant seasonal dependency. Amongst the 35 FA identified, 35–51% were saturated FA (SFA), 21–33% monounsaturated FA (MUFA), and 18–34% polyunsaturated FA (PUFA). Palmitic acid was found to be the main SFA, and was seasonal dependent, with a mean value of 30.5% for E. aeneus, 27.9% for C. taeniops, and 20.9% for S. scriba. Amongst MUFA, oleic acid, with 11–16%, was the main acid in all three species. The n6 PUFA level was low, in particular for C. taeniops (1.3–1.6%), and a little higher for S. scriba (3.6–4.2%). The three species were characterised by high amounts of n3 PUFA. Amongst them, docosa-4,7,10,13,16,19-hexaenoic acid (DHA, 22:6n3) was the highest with 5–9% for C. taeniops, 13–17% for S. scriba, and 10–16% for E. aeneus. Eicosa-5,8,11,14,17-pentaenoic acid (EPA, 20:5n3), was the second highest n3 PUFA, with 4–13%.     

Effects of trans fatty acids on markers of inflammation in bovine mammary epithelial cells

Trans fatty acids (tFA) contribute to inflammation. The objective was to investigate the effects of tFA on mRNA expression of proinflammatory markers in cultured bovine mammary epithelial cells (MAC-T cell line). Bovine mammary epithelial cells were grown in Dulbecco's Modified Eagle Medium containing 10% fetal bovine serum. Cells were then subcultured in a medium lacking fetal bovine serum, to which incremental concentrations (up to 90 μM) of elaidic acid (trans-9 C18:1) or linoleidic acid (trans-9, trans-12 C18:2) were added. Bovine serum albumin (fatty acid-free) solutions were added and cells were collected at specific time points over 48 h. Then, RNA was extracted and converted to complementary DNA for quantitative real-time PCR analysis of proinflammatory gene expression. Presence of elaidic acid caused increases in mRNA expression of interleukin (IL)-1β (3.4-fold; dose-independently over a 6-h period) and intercellular adhesion molecule (ICAM)-1 (up to 1.4-fold) relative to that for cells treated with no tFA, whereas expression of IL-6 and IL-8 was reduced 0.75- and 0.85-fold, respectively. Presence of linoleidic acid reduced mRNA expression of IL-6 and IL-8 relative to that for control (0.95- and 0.87-fold, respectively). Trans mono- and dienoic fatty acids upregulated mRNA expression of IL-1β and ICAM-1, whereas expression of IL-6 and IL-8 was downregulated in MAC-T cells. Because these genes are ultimately involved in inflammation, elaidic or linoleidic acid, either directly fed or formed in the rumen during biohydrogenation, may alter the risk for mastitis in vivo.     

Effects of fatty acid oxidation products (green odor) on rumen bacterial populations and lipid metabolism in vitro

This study investigated the effects of green odor fatty acid oxidation products (FAOP) from cut grass on lipid metabolism and microbial ecology using in vitro incubations of rumen microorganisms. These compounds have antimicrobial roles in plant defense, and we hypothesized that they may influence rumen lipid metabolism. Further, they may partially explain the higher levels of conjugated linoleic acid cis-9, trans-11 in milk from cows grazing pasture. The first of 2 batch culture experiments screened 6 FAOP (1 hydroperoxide, 3 aldehydes, 1 ketone, and 1 alcohol) for effects on lipid profile, and in particular C₁₈ polyunsaturated fatty acid biohydrogenation. Experiment 2 used the most potent FAOP to determine effects of varying concentrations and identify relationships with effects on microbial ecology. Batch cultures contained anaerobic buffer, rumen liquor, and FAOP to a final concentration of 100 μM for experiment 1. Triplicates for each compound and controls (water addition) were incubated at 39°C for 6 h. The hydroperoxide (1,2-dimethylethyl hydroperoxide, 1,2-DMEH) and the long chain aldehyde (trans-2 decenal) had the largest effects on lipid metabolism with significant increases in C_18:0 and C_18:1trans and reductions in C_12:0, C_14:0, C_16:0, C_18:1cis, C_18:2n-6, C_18:3n-3, C_20:0 and total branch and odd chain fatty acids compared with the control. This was associated with significantly higher biohydrogenation of C₁₈ polyunsaturated fatty acid. In experiment 2, 1,2-DMEH was incubated at 50, 100, and 200 μM for 2, 6, and 24 h. Increasing 1,2-DMEH concentration resulted in a significant linear increase in C_18:1trans-10, trans-11, conjugated linoleic acid, and C_18:0 and a linear decrease in C_18:2n-6 and C_18:3n-3, although the scale of this response declined with time. Microbial profiling techniques showed that 1,2-DMEH at concentrations of 100 and 200 μM changed the microbial community from as early as 2 h after addition, though microbial biomass remained similar. These preliminary studies have shown that FAOP can alter fatty acid biohydrogenation in the rumen. This change was associated with changes in the microbial population that were detected through DNA and branched- and odd-chain fatty acid profiling approaches.     

Conjugated linoleic acid production by cells and enzyme extract of Lactobacillus delbrueckii ssp. bulgaricus with additions of different fatty acids

The objective of the study was to determine the effect of fatty acid additions to the cells and enzyme extract of Lactobacillus delbrueckii ssp. bulgaricus (CCRC14009) on CLA production. Washed cells of L. delbrueckii ssp. bulgaricus, obtained by cultivation in a MRS broth, were mixed with BSA and each of the three fatty acids: linoleic, oleic, and linolenic acids in sodium phosphate buffer at pH 6.5. After incubation at 37 °C for 108 h, CLA concentration was analyzed by HPLC. Enzyme extract from the culture was also reacted with each fatty acid at 50 °C for 10 min at pH 5 to test for CLA production. Results showed that linoleic acid addition to the culture improved CLA production, indicating the presence of linoleic acid isomerase activity in the culture. The crude enzyme extract from the culture was observed to be capable of oleic and linolenic acid conversions into CLA, demonstrating the possible presence of desaturase activity in the enzyme extract.     

Trans fatty acids in a range of UK processed foods

A survey to determine the trans fatty acid content of a range of processed foods was carried out in response to recent reformulation work by the food industry to lower the artificial trans fatty acid content of processed products. Sixty two composite samples, made up of between 5 and 12 sub-samples, were collected in 2010 and were analysed for fatty acids, and a range of nutrients. The foods analysed included pizza, garlic bread, breakfast cereals, quiche, fat spreads, a range of fish and meat products, chips, savoury snacks, confectionery and ice cream. Levels of trans fatty acids were reduced considerably compared with previous UK analyses of similar foods where comparisons are possible. Concentrations of trans elaidic acid (t9-C18:1) from hydrogenated oils in all samples were <0.2 g/100 g food. These results confirm information provided by the food industry in 2007 on the levels of trans fats in key processed food sectors.     

Stability of lipids and fatty acids in frozen and gamma irradiated Patagonian toothfish (Dissostichus eleginoides) from the Southwestern Atlantic

Patagonian toothfish were captured in the Southwestern Atlantic Ocean (FAO Zone N° 41). The fatty acid profile of total lipids and the triacylglycerol and phospholipid content of control and irradiated samples (1 and 5 kGy) stored at −18 °C were analyzed at 0 and 293 days post irradiation. The fatty acids are mainly monounsaturated acids (47 g/100 g total fatty acids), the most abundant one being oleic acid (18:1 n-9). This is followed in order of abundance by saturated fatty acids (26 g/100 g total fatty acids), consisting mainly of palmitic acid (16:0). Polyunsaturated fatty acids were less abundant (17 g/100 g total fatty acids) and consisted mainly of eicosapentaeonic (20:5 n-3) and docosahexaenoic (22:6 n-3) acids. Triacylglycerol content was 563.07 mg/mL oil, whereas phospholipids were 11.21 mg/mL oil. Gamma irradiation did not significantly affect the fatty acid profile or triacylglycerol and phospholipid content of P. toothfish stored for 293 days at −18 °C. The results suggest that the species exhibits a marked stability when subjected to irradiation and prolonged storage in the frozen state.     

Evaluation of an automated hydrolysis and extraction method for quantification of total fat,lipid classes and trans fat in cereal products

The utility of an automated acid hydrolysis–extraction (AHE) system was evaluated for extraction of fat for the quantification of total, saturated, polyunsaturated, monounsaturated, and trans fat in cereal products. Oil extracted by the AHE system was assessed for total fat gravimetrically and by capillary gas chromatography (GC) for total fat, lipid classes, and trans fat. All AHE system results were compared with parallel determinations using the standard AOAC Method 996.01 or a modified version for trans fatty acids. For gravimetric and gas chromatographic evaluations, the AHE system results were equivalent to those using the standard AOAC Method (α = 0.01). Thus, the AHE oil extraction system can be used for measurement of total, saturated, polyunsaturated, monounsaturated, and trans fat with sufficient accuracy for nutrition labeling purposes, while having the advantages of reduced use of solvent, operator exposure to solvent, operator time, and potential for operator error.     

Conjugated linoleic acid (CLA) and polyunsaturated fatty acids in muscle lipids of lambs from the Patagonian area of Argentina

The concentrations of fatty acids were measured in total lipids, triacyglycerol and phospholipid fractions of intramuscular fat (IMF) from the Longissimus dorsi (LD) muscle of 10 lambs reared to approximately 30kg live weight on natural pasture with their dams. Fatty acid composition was also measured in 25 (five of each) Semitendinosus (ST), Semimembranosus (SM), Rectus femoris (RF), Gluteus (GLU) and Tensor fascia latea (TFL) muscles. Intramuscular fat percentages were similar for all muscles. Aspects of the fatty-acid patterns of relevance to human nutrition tended to favor the leg muscles with lower saturated fatty acids (SFA %), n-6/n-3 fatty acid ratios (p<0.01) and higher concentrations of the conjugated linoleic acid (CLA) (p<0.05). The estimated fatty acid concentrations (mg/100g of meat) showed higher contribution of arachidonic (C20:4 n-6), eicosapentanoic (C20:5 n-3), docosapentanoic (C22:5 n-3) and docosahexanoic (C22:6 n-3) acids in leg compared to LD lipids.