Quantitative Analysis of Nervonic and Erucic Acids in Human Milk: Comparison with Infant Formula with Different Fat Sources and Nutritional Stages.

This study aims to quantitatively analyze nervonic (C24:1 n-9) and erucic (C22:1 n-9) acids in human milk (HM) collected from Korean mothers, along with the infant formulas (IFs) with different fat sources and nutritional stages. The content of nervonic and erucic acids in HM from Korean mothers were 2.13 and 2.81 mg/100 g, accounting for 0.06 and 0.08% of the total fatty acids (FAs), respectively. These acids in IFs based on fat from plant-oil formula were higher than those in IFs based on cow milk formula. The composition of erucic acid in IFs based on cow milk formula and plant-oil formula increased with the nutritional stages, while nervonic acid only increased in IFs based on the plant-oil formula.

Development and Physicochemical Properties of Low Saturation Alternative Fat for Whipping Cream.

We developed an alternative whipping cream fat using shea butter but with low saturation. Enriched stearic-oleic-stearic (SOS) solid fat was obtained from shea butter via solvent fractionation. Acyl migration reactant, which mainly contains asymmetric SSO triacylglycerol (TAG), was prepared through enzymatic acyl migration to obtain the creaming quality derived from the ß'-crystal form. Through enzymatic acyl migration, we obtained a 3.4-fold higher content of saturated-saturated-unsaturated (SSU) TAG than saturated-unsaturated-saturated (SUS) TAG. The acyl migration reactant was refined to obtain refined acyl migration reactant (RAMR). An alternative fat product was prepared by blending RAMR and hydrogenated palm kernel oil (HPKO) at a ratio of 4:6 (w/w). The melting points, solid fat index (SFI), and melting curves of the alternative products were similar to those of commercial whipping cream fat. The alternative fat had a content of total unsaturated fatty acids 20% higher than that of HPKO. The atherogenic index (AI) of alternative fat was 3.61, much lower than those of whipping cream fat (14.59) and HPKO (1220.3), because of its low atherogenic fatty acid content and high total unsaturated fatty acids. The polymorphic crystal form determined by X-ray diffraction spectroscopy showed that the ß'-crystal form was predominant. Therefore, the alternative fat is comparable with whipping cream that requires creaming quality, and has a reduced saturated fat content.

Presence of Cholesterol in Non-Animal Organisms: Identification and Quantification of Cholesterol in Crude Seed Oil from Perilla frutescens and Dehydrated Pyropia tenera.

Studies have reported that cholesterol, a molecule found mainly in animals, is also present in some plants and algae. This study aimed to determine whether cholesterol exists in three dehydrated algae species, namely, Pyropia tenera, Saccharina japonica, and Undaria pinnatifida, and in one plant species, namely, Perilla frutescens (four perilla seed oil samples were analyzed). These species were chosen for investigation because they are common ingredients in East Asian cuisine. Gas chromatography-flame ionization detection (GC-FID) analysis found that cholesterol was present in P. tenera (14.6 mg/100 g) and in all four perilla seed oil samples (0.3-0.5 mg/100 g). High-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD) also demonstrated that cholesterol was present in P. tenera (14.2 mg/100 g) and allowed the separation of cholesterol from its isomer lathosterol. However, cholesterol could not be detected by HPLC-ELSD in the perilla seed oil samples, most likely because it is only present in trace amounts. Moreover, liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed the presence of cholesterol in both P. tenera and perilla seed oil. MRM results further suggested that lathosterol (a precursor of cholesterol) was present in P. tenera.

In Vitro Study for Lipolysis of Soybean Oil, Pomegranate Oil, and Their Blended and Interesterified Oils under a pH-Stat Model and a Simulated Model of Small Intestinal Digestion.

In this study, two in vitro digestion models were employed to compare the rate of lipolysis in soybean oil (SBO), pomegranate oil (PGO), a physical blend (PHY, 1:1 molar ratio of SBO:PGO, w/w), and their enzymatically interesterified oil (IO). In the pH-stat digestion model (emulsified oils with bile salts), PGO emulsion containing 74.7% conjugated form of linolenic acid (CLn) showed a significantly lower release rate of free fatty acid (FFA) than the other oil emulsions (p < 0.05). In FFA release rates and oil droplet sizes between PHY and IO emulsions, no significant differences were observed (p > 0.05). In a simulated model of small intestinal digestion, the lipolysis rates of SBO, PGO, PHY, and IO after digestion for 30 min in digestion fluids were 80.4%, 66.5%, 74.8%, and 77.0%, respectively. The rate of lipolysis in PGO was significantly lower than that in SBO (p < 0.05), and the lowest lipolysis rate was observed in the conjugated form of trilinolenoyl glycerol (CLn-CLn-CLn).

Physicochemical characteristics of fat blend from hydrogenated coconut oil and acyl migrated palm mid-fraction.

Palm mid-fraction (PMF), which has a high content of symmetric POP, was converted to asymmetric PPO (APMF) via acyl migration. After solvent fractionation, the liquid phase of acyl migrated PMF (APMF-L) was obtained and blended with hydrogenated coconut oil (HCO, 50:50, w/w) to produce a fat blend (namely, an alternative fat blend) which had reduced saturated fatty acid content while having similar melting behavior to HCO. In an alternative fat blend, the major fatty acids were lauric (27.94), palmitic (26.93) and oleic (15.75 mol%) acid. The solid fat index was quite similar to that of HCO, especially at 28-44 °C. Nevertheless, an alternative fat blend had lower saturated fatty acid content, by 18%, compared to HCO. The content of highly atherogenic myristic acid was reduced by approximately 40%. The alternative fat blend in this study could be used as a raw material for non-dairy cream with low saturated fat content.

Oxidation Stability of O/W Emulsion Prepared with Linolenic Acid Enriched Diacylglycerol.

The sn-1,3-regiospecific Rhizomucor miehei lipase (Lipozyme RM IM) was employed to produce structured diacylglycerol (SL-DAG), which contained 67.3 mol% DAG with 27.2 area% of C18:3. To investigate the oxidative stability of the SL-DAG in emulsion form, 5% oil-in-water (O/W) emulsions were prepared with 200 and 400 ppm sinapic acid. It was shown that the hydroperoxide values of the control (without any antioxidant) was the highest (117.7 meq/L) on day 43 of storage and thereafter the value decreased. However, the emulsions with 200 and 400 ppm sinapic acid resulted in slow oxidation degree until day 64 of storage (30.3 and 7.3 meq/L, respectively). Aldehyde measurements for the 200 ppm sinapic acid emulsion (12.8 mmol/mol) and the 400 ppm sinapic acid emulsion (7.5 mmol/mol) also showed better oxidative stability than that for the 200 ppm catechin emulsion (27.4 mmol/mol) and the control (52.7 mmol/mol). Although the SL-DAG in the emulsions contains high levels of polyunsaturated fatty acids, the degree of oxidation in the emulsions can be reduced when sinapic acid is used as an antioxidant.

Effects of Caffeic Acid Phenethyl Ester and 4-Vinylcatechol on the Stabilities of Oil-in-Water Emulsions of Stripped Soybean Oil.

Caffeic acid phenethyl ester (CAPE) and 4-vinylcatechol (4-VC) were prepared for studying their antioxidative activities in emulsion. Oil-in-water emulsions of stripped soybean oil containing 200 ppm of CAPE, 4-VC, or α-tocopherol were stored at 40 °C in the dark for 50 days, and proton nuclear magnetic resonance ((1)H NMR) was used to identify and quantify the oxidation products. Emulsion droplet sizes, peroxide values, and levels of primary oxidation products (i.e., hydroperoxides) and secondary oxidation products (i.e., aldehydes) were determined. The results showed that CAPE (200 ppm) and 4-VC (200 ppm) had significantly greater antioxidant activities on the oxidation of stripped soybean oil-in-water emulsions than α-tocopherol (200 ppm). The peroxide values of CAPE (8.4 mequiv/L emulsion) and 4-VC (15.0 mequiv/L emulsion) were significantly lower than that of α-tocopherol (33.4 mequiv/L emulsion) (p < 0.05) on 36 days. In addition, the combinations of CAPE + α-tocopherol (100 + 100 ppm) or 4-VC + α-tocopherol (100 + 100 ppm) had better antioxidant activities than α-tocopherol (200 ppm). For CAPE + α-tocopherol, 4-VC + α-tocopherol, and α-tocopherol, the amounts of conjugated diene forms were 16.67, 13.72, and 16.32 mmol/L emulsion, and the concentrations of aldehydes were 2.15, 1.13, and 4.26 mmol/L emulsion, respectively, after 50 days of storage.

Relative oxidative stability of diacylglycerol and triacylglycerol oils.

To compare the oxidative stability between diacylglycerol (DAG) oil and conventional triacylglycerol (TAG) oil (that is, soybean oil), the prepared stripped diacylglycerol oil (SDO) and soybean oil (SSBO) were stored at 60 °C in the dark for 144 h. During storage peroxide values (POVs), contents of aldehydes, unsaturated fatty acids were measured to evaluate the oxidative stabilities of the 2 oils. The results showed the content of C18:2, C18:3, and total unsaturated fatty acid decreased faster in DAG oil than in soybean oil, whereas the decreased rate of C18:1 was similar in 2 oils. Also, both rate constants (K1 and K2) obtained from POV (K1 ) and total aldehydes (K2 ) indicated that DAG oil (K1 = 3.22 mmol/mol FA h(-1) , K2 = 0.023 h(-1)) was oxidized more rapidly than soybean oil (K1 = 2.56 mmol/mol FA h(-1) , K2 = 0.021 h(-1)), which was mainly due to the difference of acylglycerol composition of the 2 oils along with higher C18:3 (9.6%) in SDO than SSBO (5.7%). It is concluded that DAG was more easily oxidized than soybean oil at 60 °C in the dark for 144 h.

Effects of flavonoid glycosides obtained from a Ginkgo biloba extract fraction on the physical and oxidative stabilities of oil-in-water emulsions prepared from a stripped structured lipid with a low omega-6 to omega-3 ratio.

In this study, we have produced a structured lipid with a low ω6/ω3 ratio by lipase-catalysed interesterification with perilla and grape seed oils (1:3, wt/wt). A Ginkgo biloba leaf extract was fractionated in a column packed with HP-20 resin, producing a flavonoid glycoside fraction (FA) and a biflavone fraction (FB). FA exhibited higher antioxidant capacity than FB, showing 58.4 mmol gallic acid equivalent (GAE)/g-of-total-phenol-content, 58.8 mg quercetin equivalent (QUE)/g-of-total-flavonoid-content, 4.5 mmol trolox/g-of-trolox-equivalent antioxidant capacity, 0.14 mg extract/mL-of-free-radical-scavenging-activity (DPPH assay, IC50), and 2.3 mmol Fe2SO4 · 7H2O/g-of-ferric-reducing-antioxidant-power. The oil-in-water emulsion containing the stripped structured lipid as an oil phase with FA exhibited the highest stability and the lowest oil globule diameters (d43 and d32), where the aggregation was unnoticeable by Turbiscan and particle size analyses during 30 days of storage. Furthermore, FA was effective in retarding the oxidation of the emulsions.

Preparation of recombined milk using modified butterfats containing α-linolenic acid.

UNLABELLED: Modified butterfats (MBFs) were produced by lipase-catalyzed interesterification with 2 substrate blends (6:6:8 and 4:6:10, by weight) of anhydrous butterfat (ABF), palm stearin, and flaxseed oil in a stirred-batch type reactor after short path distillation. The 6:6:8 and 4:6:10 MBF contained 21.7% and 26.5%α-linolenic acid, respectively. Total saturated fatty acids of the MBFs ranged from 41.4% to 47.4%. The cholesterol contents of the 6:6:8 and 4:6:10 MBFs were 21.0 and 12.1 mg/100 g, respectively. In addition, the melting points of the 6:6:8 and 4:6:10 MBFs were 32 °C and 31 °C, respectively. After preparation of recombined milks (oil-in-water emulsions) with MBFs, the stability of emulsions prepared with the MBFs (6:6:8 and 4:6:10) was compared to those with ABF during 10-d storage at 30 °C. Skim milk powder (containing 1% protein) was added to prepare emulsions as an emulsifier. Microstructures of emulsions freshly prepared with the ABF and the MBFs consisted of uniform fat globules with no flocculation during 10-d storage. With respect to fat globule size distribution, the volume-surface mean droplet diameter (d(32)) of the 6:6:8 and 4:6:10 MBF emulsions ranged between 0.33 and 0.34 µm, which was similar to the distribution in ABF emulsion. PRACTICAL APPLICATION: Milk, an expensive dairy food, has been widely used in various milk-derived food products. Modified butterfats (MBFs) contain α-linolenic acid as an essential fatty acid. Emulsion stability of recombined milks (oil-in-water emulsions) with MBFs was similar to that in anhydrous butterfat emulsion during 10-d storage. They may be a promising alternative for reconstituted milks to use in processed milk-based products.

Antioxidative activities of Ginkgo biloba extract on oil/water emulsion system prepared from an enzymatically modified lipid containing alpha-linolenic acid.

UNLABELLED: The desired mix of alpha-linolenic acid (ALA)-enriched structured lipid (SL) and physically blended lipid (PB) was prepared from grape seed oil and perilla oil at a weight ratio of 3:1. The major triacylglycerol species (LnLnL) in PB was drastically increased after interesterification (SL), from 0.5% to 16.8%. After the reaction, the total unsaturated fatty acid at the sn-2 position was decreased from 98.83% in PB to 91.36% in SL. The reduction of vitamin E compounds was also observed. Compared with a PB-based emulsion, SL-based emulsions showed oxidative instability, as assessed by lipid hydroperoxide (LOOH) and 2-thiobarbituric acid-reactive substances (TBARS) values, which was mainly due to the SL which contained less LA, ALA, and ΣUSFA at the sn-2 position and less γ-tocopherol than did PB. PB-, and SL-based emulsions with Ginkgo biloba extract (GBE) which showed significantly lower values of LOOH and TBARS compared to a blank control. GBE was effective in retarding the oxidation of the emulsion by quenching the free radicals in the water phase of the emulsion and inhibiting the formation of primary and secondary oxidation products. These results indicate that GBE could be used as an antioxidant additive for stabilizing ALA-enriched emulsions. PRACTICAL APPLICATION: The results suggest the possibility to supplement Ginkgo biloba extract in alpha linolenic acid-enriched structured lipid-based emulsions which would increase the therapeutic value and enhance the antioxidant potential of the emulsions.

Studies of reaction variables for lipase-catalyzed production of alpha-linolenic acid enriched structured lipid and oxidative stability with antioxidants.

Alpha-linolenic acid (ALA) enriched structured lipid (SL) was produced by lipase-catalyzed interesterification from perilla oil (PO) and corn oil (CO). The effects of different reaction conditions (substrate molar ratio [PO/CO 1:1 to 1:3], reaction time [0 to 24 h], and reaction temperature [55 to 65 °C]) were studied. Lipozyme RM IM from Rhizomucor miehei was used as biocatalyst. We obtained 32.39% of ALA in SL obtained under the optimized conditions (molar ratio-1:1 [PO:CO], temperature-60 °C, reaction time-15 h). In SL, the major triacylglycerol (TAG) species (linolenoyl-linolenoyl-linolenoyl glycerol [LnLnLn], linolenoyl-linolenoyl-linoleoyl glycerol [LnLnL]) mainly from PO and linoleoyl-linoleoyl-oleoyl glycerol (LLO), linoleoyl-oleoyl-oleoyl glycerol (LOO), palmitoyl-linoleoyl-oleoyl glycerol (PLO) from CO decreased while linolenoyl-linolenoyl-oleoyl glycerol (LnLnO) (18.41%), trilinolein (LLL) (9.06%), LLO (16.66%), palmitoyl-linoleoyl-linoleoyl glycerol (PLL) (9.69%) were increased compared to that of physical blend. Total tocopherol content (28.01 mg/100 g), saponification value (SV) (192.2), and iodine value (IV) (161.9) were obtained. Furthermore, oxidative stability of the SL was also investigated by addition of 3 different antioxidants (each 200 ppm of rosemary extract [SL-ROS], BHT [SL-BHT], catechin [SL-CAT]) was added into SL and stored in 60 °C oven for 30 d. 2-Thiobabituric acid-reactive substances (TBARS) value was 0.16 mg/kg in SL-CAT and 0.18 mg/kg in SL-ROS as compared with 0.22 mg/kg in control (SL) after oxidation. The lowest peroxide value (POV, 200.9 meq/kg) and longest induction time (29.88 h) was also observed in SL-CAT.

Oxidative comparison of emulsion systems from fish oil-based structured lipid versus physically blended lipid with purple-fleshed sweet potato (Ipomoea batatas L.) extracts.

The effects of the purple-fleshed sweet potato extract (PFSPE) on oxidation stabilities of a model oil-in-water emulsion prepared with enzymatically synthesized fish oil-soybean oil structured lipid (SL) versus physically blended lipid (PBL) without modification were evaluated. The anthocyanins in PFSPE were analyzed and identified by HPLC-MS. The fatty acid composition of SL was similar to that of PBL, except palmitic acid (1.48 in PBL and 9.61% in SL) and linoleic acid (62.47 in PBL and 49.58% in SL). Peonidin 3-caffeoylsophoroside-5-glucoside, peonidin-3-(6',6'-caffeoylferuloylsophoroside)-5-glucoside, peonidin-dicaffeoylsophoroside-5-glucoside, peonidin 3-(6',6"-caffeoyl-p-hydroxybenzoylsophoroside)-5-glucoside were identified as the major anthocyanin compounds in PFSPE. Different levels (200, 500, 1000 ppm) of PFSPE were added into both SL- and PBL-based emulsions, with 200 ppm catechin as comparison. Oxidation was monitored by measuring the peroxide value and thiobarbituric acid reactive substances. The antioxidant activity of PFSPE increased with an increased concentration, the concentration of 1000 ppm showed high antioxidant ability similar to that of catechin in both PBL- and SL-based oil-in-water emulsions. It is notable that the SL-based emulsion appeared to have better oxidative stability than the PBL-based emulsion.

Production of trans-free margarine stock by enzymatic interesterification of rice bran oil, palm stearin and coconut oil.

BACKGROUND: Trans-free interesterified fat was produced for possible usage as a spreadable margarine stock. Rice bran oil, palm stearin and coconut oil were used as substrates for lipase-catalyzed reaction. RESULTS: After interesterification, 137-150 g kg(-1) medium-chain fatty acid was incorporated into the triacylglycerol (TAG) of the interesterified fats. Solid fat contents at 25 degrees C were 15.5-34.2%, and slip melting point ranged from 27.5 to 34.3 degrees C. POP and PPP (beta-tending TAG) in palm stearin decreased after interesterification. X-ray diffraction analysis demonstrated that the interesterified fats contained mostly beta' polymorphic forms, which is a desirable property for margarines. CONCLUSIONS: The interesterified fats showed desirable physical properties and suitable crystal form (beta' polymorph) for possible use as a spreadable margarine stock. Therefore, our result suggested that the interesterified fat without trans fatty acid could be used as an alternative to partially hydrogenated fat.

Enrichment of pinolenic acid at the sn-2 position of triacylglycerol molecules through lipase-catalyzed reaction.

Reports have shown that Delta-5 polyunsaturated fatty acids (-5 PUFA) are enriched at sn-1,3 positions of triacylglycerols (TAG) in pine (Pinus koraiensis) nut oil (Pn). As a major Delta-5 PUFA, pinolenic acid (Pi) is about 14.2% in the oil, while the percentage of Pi at the sn-1 and/or sn-3 positions in TAG was found more than 20%. In this current study, the enhancement of Pi at the sn-2 position has been achieved by acyl migration during the lipase-catalyzed inter-esterification between Pn and palm stearin (Ps). After reaction, the proportion of Pi increased at sn-2 positional fatty acid even is similar to that in total fatty acid; for example, in the inter-esterified product of 50:50 (Pn:Ps), the same amount of Pi (7.1%) present was detected both at the sn-2 and sn-1,3 positions. However, the reduction of phytosterols and tocopherols are observed in the inter-esterified products.

Production and physicochemical properties of functional-butterfat through enzymatic interesterification in a continuous reactor.

Modified-butterfat (MBF) was synthesized with four blends (8:6:6, 6:6:8, 6:6:9, and 4:6:10, by weight) of anhydrous butterfat (ABF), palm stearin (PS) and flaxseed oil (FSO) through enzymatic interesterification in a continuous packed-bed reactor. Flow rate effect of 3, 5, 8 and 10 mL/min on enzymatic interesterification was investigated. By increasing the enzyme contact time with substrates (decreased flow rates), not only did melting and crystallization points shift to lower temperature but also the equivalent carbon number, ECN 36-38 from FSO decreased. Further all reactions were performed at flow rate of 5 mL/min (contact time 140 min) in a continuous reactor packed with 150 g of Lipozyme RM IM. After short path distillation, alpha-linolenic acid composition (%) of 8:6:6, 6:6:8, 6:6:9, and 4:6:10 MBFs were 16, 21, 23 and 25%, respectively. The contents of ECN 36-38, and ECN 48-50 decreased in the blends and MBFs for each substrate ratio. ECN 42-46 in the newly produced TAG increased. Melting points of MBFs were 38 degrees C (8:6:6), 35.5 degrees C (6:6:8), 34 degrees C (6:6:9), and 32 degrees C (4:6:10). MBFs interesterified with FSO contained phytosterols (17-36 mg/100 g) and tocopherols (116-173 microg/g). The products of 8:6:6, 6:6:8, 6:6:9 and 4:6:10 MBFs were softer (69, 88, 80, and 92%, respectively) than pure butterfat at refrigeration temperature. The polymorphic form changed from beta form (blends) to desirable crystalline structure of beta' form (MBFs). Crystal morphology of MBFs also changed and was composed of small spherulites of varying density.

Development and characterization of structured lipids containing capric and conjugated linoleic acids as functional dietary lipid molecules.

Recently, dietary oil with high diacylglycerol (DAG) contents, so called DAG-oil, was introduced in Japan and the USA. It was claimed that the oil mostly composed of DAG is metabolized differently from conventional triacylglycerol oil, reducing body weight and fat mass because DAG tends to be oxidized to provide energy rather than stored as fat in the body. Monoacylglcyerol and DAG could be prepared by lipase-catalyzed reactions including hydrolysis, esterification, and glycerolysis. In this study, modified lipid containing some DAG esterified with the health-beneficial medium-chain fatty acids and conjugated linoleic acid was produced by lipase-catalyzed reactions. Many health benefits of medium-chain fatty acids (C6:0-C12:0) and conjugated linoleic acid isomers have been reported, including anticarcinogenic and antiatherogenic activities, and being rapid energy sources for humans with little or no deposition as body fat. The produced lipid molecules in this study have potential applications as functional healthy dietary fats and oils.