Study on the characteristics of glycerides and phospholipids in human milk from Tibet.

The unique geographical characteristics and food culture of Tibet can affect the nutrition of human milk lipids. But little has been done in the comparison of the lipids between Tibet and other areas. This study gives in-depth analysis of the species, concentration and composition of lipid subclasses at the molecular level of the Tibetan human milk. There were averagely 132 ± 30 species of lipids, among which triglycerides (TAGs), phosphatidylethanolamine (PE) and sphingomyelin (SM) accounted for 79.78% of the total species number in the Tibetan human milk samples. The contents of TAG, SM, phosphatidylcholine (PC), and PE in the Tibetan human milk were 85.84%, 17.79%, 25.94% and 55.81% of those in the comparative human milk of China, respectively. The contents of TAGs and diglycerides (DAGs) with PUFAs in Tibetan human milk were significantly lower than those in the comparative group. However, the content and percentage of TAGs and DAGs with odd-chain saturated fatty acids were both higher in the Tibetan human milk than those in the comparative human milk. In total, 18 molecular species of lipids were downregulated and 5 ones were upregulated in the Tibetan human milk compared with those in the comparative human milk of China. The profile of lipids in the Tibetan human milk at the molecular level provided the scientific basis for maternal diet and supplemented the Chinese human milk lipids database.

Identification of Glyceryl MonoRicinoleate (GMR) isomers using RP-HPLC and regio-specificity of lipases.

In the present study, we report a reverse-phase high-performance liquid chromatography (RP-HPLC) method for separation of the regio-isomers of Glyceryl MonoRicinoleate (GMR) identified using position specificity of lipases. The approaches explored to identify these regio-isomers include LC-mass spectrometry, UV spectroscopy, and selective hydrolysis with lipases. A distinct UV absorption spectrum and λmax values for each isomer were noted, and mass spectral analysis further revealed their molecular weight. Lastly, the purified regio-isomers were subjected to hydrolysis with two distinctive regio-specific lipases to identified as sn-2 and sn-1(3) GMR. The current methodology of using analytic tool and enzyme specificity provides a useful platform for identifying regio-isomers for structured lipid synthesis.

The Lipid and Glyceride Profiles of Infant Formula Differ by Manufacturer, Region and Date Sold.

We tested the hypothesis that the lipid composition of infant formula is consistent between manufacturers, countries and target demographic. We developed techniques to profile the lipid and glyceride fraction of milk and formula in a high throughput fashion. Formula from principal brands in the UK (2017-2019; bovine-, caprine-, soya-based), the Netherlands (2018; bovine-based) and South Africa (2018; bovine-based) were profiled along with fresh British animal and soya milk and skimmed milk powder. We found that the lipid and glyceride composition of infant formula differed by region, manufacturer and date of manufacture. The formulations within some brands, aimed at different target age ranges, differed considerably where others were similar across the range. Soya lecithin and milk lipids had characteristic phospholipid profiles. Particular sources of fat, such as coconut oil, were also easy to distinguish. Docosahexaenoic acid is typically found in triglycerides rather than phospholipids in formula. The variety by region, manufacturer, date of manufacture and sub-type for target demographics lead to an array of lipid profiles in formula. This makes it impossible to predict its molecular profile. Without detailed profile of the formula fed to infants, it is difficult to characterise the relationship between infant nutrition and their growth and development.

Interesterification of rice bran wax and palm olein catalyzed by lipase: Crystallization behaviours and characterization.

Rice bran wax (RBW) is a traditional plant based natural wax and an increasingly popular component in textiles, fruit coatings and cosmetics. Properties of RBW can be modified by acyglycerols, and the resulting products can possess features with great potential in different applications. In this study, RBW was interesterified with palm olein (POL) catalyzed by Lipozyme TL IM, and the effects of RBW on the crystallization rate, solid fat content (SFC) and thermodynamic properties were investigated. The crystallization rates of RBW-based enzymatically interesterified (EIE) products were significantly higher than both the starting mixture and fully hydrogenated rapeseed oil (FHRSO). The EIE RBW-based samples were predominantly crystallized in ß' form, and presented a much smoother SFC profile as compared to physically blended raw materials. The SFC values were significantly decreased, conversely increased, and remained constant, and at 10 °C, 20-30 °C, and 35-40 °C as the wax ester and acylglycerols compositions changes. Overall, RBW-based samples after EIE showed an increased hardness and good surface properties, which make it a potential plastic fats substitute.

Simultaneous high-temperature gas chromatography with flame ionization and mass spectrometric analysis of monocarboxylic acids and acylglycerols in biofuels and biofuel intermediate products.

Triacyl-, diacyl- and monoacylglycerols (TAGs, DAGs, MAGs) along with monocarboxylic acids (MCAs) are intermediate products in many triacylglycerol oil-to-biofuel conversion pathways. Accumulation of these compounds leads to poor biofuel characteristics and may result in fuel system damage. We developed a method for simultaneous identification and quantification of a wide range of MCAs (C4-C18), MAGs, DAGs, and TAGs. The method is based on trimethylsilylation followed by high temperature GC with programmed temperature vaporizer (PTV) injection coupled to parallel FID and MS detectors (HTGC-FID/MS). To minimize the discrimination of both low and high molecular weight species typically occurring on the injector, we optimized injection conditions using a central composite design. The critical variables were the time at initial temperature (40 °C), splitless time, and the interaction between these two parameters. Among three tested electron ionization source/quadrupole analyzer temperatures, a 350/200 °C setting provided the highest response and signal-to-noise ratio for TAGs and did not have an effect on MAGs and DAGs. Similar results were obtained when quantifying target analytes in intermediate products of soybean oil cracking with FID and MS (using specific acylglycerol fragmentation ions). The instrumental FID limits of detection (LODs) were 0.07-0.27 ng for most of the target analytes. Selected ion monitoring (SIM) LODs were 0.01-0.05 ng for MCAs and 0.03-0.14 ng for acylglycerols. For the total ion current (TIC), LODs observed increased with acyl chain length and degree of unsaturation, resulting in an increase from 0.05 to 0.18 ng for MCAs (C5 to C18) and from 0.03 to 1.8 ng for acylglycerols (TAGs C8 to C22). Deviations in the repeatability of sample preparation, intra- and inter-day analyses, including sample stability over an eight-day time period, did not exceed 10% variance. These results demonstrate that the developed method is accurate and robust for the determination of acylglycerols and MCAs produced during the processing of TAGs into biofuels.

Rapid and direct determination of fatty acids and glycerides profiles in Schisandra chinensis oil by using UPLC-Q/TOF-MSE.

Fatty acids and glycerides are globally accepted quality and nutrition indicators of oils. Schisandra chinensis (S. chinensis) is a good functional oil source, with an oil content of 10-50% (dry weight). In this study, the UPLC-Q/TOF-MSE technique was developed to profile FFA and glycerides in the S. chinensis oils directly. The results showed that all of the 36 FFA calibration equations of the mixture standard had good linear relationships (R2 > 0.99). The limit of detection for the tested compounds ranged from 0.0001 to 0.0200 µg/mL, while the limit of quantification ranged from 0.0005 to 0.1300 µg/mL. In total, seventeen FFAs, six diglycerides and 20 triglycerides were identified. Linoleic, oleic, stearic and palmitic acids were the most abundant FFAs in the S. chinensis oils. It was also found that S. chinensis oil is rich in the L-L, L-L-L, O-L-L and O-L-O glycerides. These results will be helpful for the use of this technique in physicochemical evaluation and for further application development.

Characterization, anti-oxidative effect of grape seed powder and in silico affinity profiling of polyphenolic and extra-phenolic compounds for calpain inhibition.

Vitis vinifera grape is a highly cultivated crop and solid wastes generated by the wine industry are largely under exploited. Plentiful studies have intended analyzing the polyphenolic content of grape seeds but characterization of non phenolic compounds is rather scarce. The present study aimed at the selective extraction of lipid, phenolic and aqueous phases from grape seed powder (GSP) in order to establish their intimate composition, as well as their antioxidant and chelating properties underlying partly their biological effects. Major non phenolic compounds identified in the lipid phase were glyceryl-monostearate and 2-monostearin whereas fructofuranose and sucrose were the most abundant in the aqueous phase. Among the most abundant compounds detected in the various phases, the polyphenol quercetin exhibited the best affinity and free binding energy towards the active site of the calcium-dependent protease calpain. Polyphenols likely constitute the bioactive part of GSP that should be exploited as safe modulators of intracellular signaling which is likely at the basis of their health beneficial effects. Nevertheless other compounds as lipids or sugars should be valorized along with polyphenols to improve their bioavailability into highly protected organs as brain or eye.

Modifications of Atlantic salmon by-product oil for obtaining different ω-3 polyunsaturated fatty acids concentrates: An approach to comparative analysis.

Omega-3 polyunsaturated fatty acids (ω-3 PUFAs) rich 2-monoacylglycerols (2-MAG), omega-3 polyunsaturated free fatty acids (ω-3 PUFFAs) concentrate, and PUFA enriched acylglycerols were prepared from salmon frame bone oil (SFBO) by enzymatic alcoholysis, urea complexation, and enzymatic esterification, respectively. The yields of 2-MAG, ω-3 PUFFAs concentrate, and PUFA enriched acylglycerols were 40.25, 16.52, and 15.65%, respectively. ω-3 PUFFAs concentrate and PUFA enriched acylglycerols showed darker red color than SFBO and 2-MAG due to aggregation of astaxanthin pigment in ω-3 PUFFAs concentrate during urea complexation. The viscosity and specific gravity of SFBO and PUFA enriched acylglycerols showed similar values whereas 2-MAG and ω-3 PUFFAs showed significantly (p < 0.05) lower values. Stability parameters like acid value, peroxide value, free fatty acid value, and p-anisidine value of SFBO and ω-3 PUFAs concentrates were within acceptable limits except extreme high acid value and free fatty acid value of ω-3 PUFFAs concentrate. Thermogravimetric analysis showed similar and higher thermal stability of SFBO and PUFA enriched acylglycerols than 2-MAG and ω-3 PUFFAs concentrate. The ω-3 PUFAs content in 2-MAG, ω-3 PUFFAs concentrate, and PUFA enriched acylglycerols was increased to 20.81, 52.96, and 51.74% respectively from 13.54% in SFBO. ω-3 PUFFAs concentrate and PUFA enriched acylglycerols showed higher DPPH and ABTS radical scavenging activity than SFBO and 2-MAG. The results obtained from this study suggest the production of PUFA enriched acylglycerols rich in ω-3 PUFAs supplements from fish oil for human and pet animals.

Effects of fatty acid glyceride product SILOhealth 104 on the growth performance and carcass composition of broiler chickens.

Butyric acid is the primary energy source for colonocytes, and has shown potential as an alternative to in-feed antibiotics, due to its antimicrobial activity and positive effects on production performance traits of broiler chickens. SILOhealth 104 (SILO S.P.A., Florence, Italy) is a commercial product mainly containing mono- and di-glycerides of butyrate with a small portion of propionic, caprylic, capric, and lauric acid mono- and di-glycerides. Its effects on broiler performance and carcass composition have yet to be evaluated. Four-hundred-eighty day-old male Ross 308 birds were divided into different dietary treatment groups with equal starting weights and fed a diet containing 0, 500, 1,000, 2,000, or 3,000 ppm of SILOhealth 104 for 35 days. There were no significant differences in overall average daily gain or feed: gain ratio with the addition of SILOhealth 104 to the diets (P > 0.05). At 5 wk of age, abdominal fat weight was reduced in birds supplemented with SILOhealth 104 in a dose-responsive manner (P < 0.05), while breast muscle weight increased with supplementation, with significant increases in 2,000 ppm and 3,000 ppm birds compared to controls (P < 0.05). A significant reduction in gene expression of both forkhead box protein O4 and myostatin, 2 factors that can inhibit protein synthesis, was found in the breast muscle of all SILOhealth 104 treated birds (P < 0.05). In addition, gene expression in the adipose tissue, including acetyl-CoA carboxylase alpha and lipoprotein lipase, which are associated with lipid metabolism, was significantly decreased and increased, respectively, by the supplementation of SILOhealth 104 (P < 0.05). These data suggest that the components of SILOhealth 104 can positively affect the deposition of muscle, while reducing abdominal fat deposition in broiler chickens.

Ultrasound Assisted Synthesis of Hydroxylated Soybean Lecithin from Crude Soybean Lecithin as an Emulsifier.

Soybean lecithin is a by-product obtained during degumming step of crude soybean oil refining. Crude soybean lecithin (CSL) contains major amount of phospholipids (PLs) along with minor amount of acylglycerols, bioactive components, etc. Due to presence of PLs, CSL can be used as an emulsifier. Crude soybean lecithin (CSL) was utilized to synthesize hydroxylated soybean lecithin (HSL) by hydroxylation using hydrogen peroxide and catalytic amount of lactic acid to enhance the hydrophilicity and emulsifying properties of CSL. To reduce the reaction time and to increase rate of reaction, HSL was synthesized under ultrasound irradiation. The effect of different operating parameters such as lactic acid, hydrogen peroxide, temperature, ultrasonic power and duty cycle in synthesis of HSL were studied and optimized. The surface tension (SFT), interfacial tension (IFT) and the critical micelle concentration (CMC) of the HSL (26.11 mN/m, 2.67 mN/m, 112 mg/L) were compared to CSL (37.53 mN/m, 6.22 mN/m, 291 mg/L) respectively. The HSL has better emulsion stability and low foaming characteristics as compared to CSL. Therefore, the product as an effective emulsifier can be used in food, pharmacy, lubricant, cosmetics, etc.

Production of Biodiesel from Acid Oil via a Two-Step Enzymatic Transesterification.

A two-step enzymatic transesterification process in a solvent-free system has been developed as a novel approach to the production of biodiesel using acid oil from rice bran oil soapstock. The acid oil consisted of 53.7 wt% fatty acids, 2.4 wt% monoacylglycerols, 9.1 wt% diacylglycerols, 28.8 wt% triacylglycerols, and 6.0 wt% others. Three immobilized lipases were evaluated as potential biocatalysts, including Novozym 435 from Candida antarctica, Lipozyme RM IM from Rhizomucor miehei, and Lipozyme TL IM from Thermomyces lanuginosus. The effects of molar ratio of acid oil to ethanol, temperature, and enzyme loading were investigated to determine the optimum conditions for the transesterification with the three immobilized lipases. The optimum conditions of the three immobilized lipases were a molar ratio of 1:5 (acid oil to ethanol), the temperature range of 30-40°C, and the enzyme loading range of 5-10%. The two-step transesterification was then conducted under the optimum conditions of each lipase. The stepwise use of Novozym 435 and Lipozyme TL IM or Lipozyme RM IM and Lipozyme TL IM resulted in similar or higher levels of yield to the individual lipases. The maximum yields obtained in both stepwise uses were ca. 92%.

Identification and characterization of phenolic compounds in hydromethanolic extracts of sorghum wholegrains by LC-ESI-MS(n).

Hydromethanolic extracts of brown, red, and white sorghum whole grains were analysed by LC-MS(n) in negative ESI mode within the range m/z 150-550amu. Besides the flavonoids already reported in sorghum, a number of flavonoids were also identified in the sorghum grain for the first time, including flavanones, flavonols and flavanonols, and flavan-3-ol derivatives. Various phenylpropane glycerides were also found in the sorghum grain, the majority of them are reported here for the first time, and a few of them were detected with abundant peaks in the extracts, indicating they are another important class of phenolic compounds in sorghum. In addition, phenolamides were also found in sorghum grain, which have not been reported before, and dicaffeoyl spermidine was detected in high abundance in the extracts of all three type sorghum grains. These results confirmed that sorghum is a rich source of various phenolic compounds.

Formation and reduction of 3-monochloropropane-1,2-diol esters in peanut oil during physical refining.

In the present study, lab-scale physical refining processes were investigated for their effects on the formation of 3-monochloropropane-1,2-diol (3-MCPD) esters. The potential precursors, partial acylglycerols and chlorines were determined before each refining step. 3-MCPD esters were not detected in degummed and bleached oil when the crude oils were extracted by solvent. While in the hot squeezed crude oils, 3-MCPD esters were detected with low amounts. 3-MCPD esters were generated with maximum values in 1-1.5h at a certain deodorizing temperature (220-260°C). Chlorine seemed to be more effective precursor than partial acylglycerol. By washing bleached oil before deodorization with ethanol solution, the precursors were removed partially and the content of 3-MCPD esters decreased to some extent accordingly. Diacetin was found to reduce 3-MCPD esters effectively.

Use of re-esterified palm oils, differing in their acylglycerol structure, in weaning-piglet diets.

Re-esterified oils are new fat sources obtained from chemical esterification of acid oils with glycerol (both economically interesting by-products from oil refining and biodiesel industries, respectively). The different fatty acid (FA) positional distribution and acylglycerol composition of re-esterified oils may enhance the apparent absorption of saturated fatty acids (SFA) and, thus, their overall nutritive value. The aim of the present study was to investigate the potential use of re-esterified palm oils, in comparison with their corresponding acid and native oils, and also with an unsaturated fat source in weaning-piglet diets. The parameters assessed were: FA apparent absorption, acylglycerol and free fatty acid (FFA) composition of feces, and growth performance. One-hundred and twenty weaning piglets (average weight of 8.50±1.778 kg) were blocked by initial BW (six blocks) and randomly assigned to five dietary treatments, resulting in four piglets per pen and six replicates per treatment. Dietary treatments were a basal diet supplemented with 10% (as-fed basis) of native soybean oil (SN), native palm oil (PN), acid palm oil (PA), re-esterified palm oil low in mono- (MAG) and diacylglycerols (DAG) (PEL), or re-esterified palm oil high in MAG and DAG (PEH). Results from the digestibility balance showed that SN reached the greatest total FA apparent absorption, and statistically different from PN, PA and PEL (P0.05), but PEH achieved the greatest total FA apparent absorption. Animals fed PEL, despite the fact that PEL oil contained more sn-2 SFA, did not show an improved absorption of SFA (P>0.05). Animals fed PA and PN showed similar apparent absorption coefficients (P>0.05), despite the high FFA content of PA oil. The acylglycerol and FFA composition of feces was mainly composed of FFA. There were no significant differences in growth performance (P>0.05). Results of the present study suggest that, despite the different acylglycerol structure of re-esterified oils, there were no significant differences in digestibility or performance with respect to their corresponding PN and PA oils in weaning-piglet diets.

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.

Influence of oil composition on the formation of fatty acid esters of 2-chloropropane-1,3-diol (2-MCPD) and 3-chloropropane-1,2-diol (3-MCPD) under conditions simulating oil refining.

The toxicological relevance and widespread occurrence of fatty acid esters of 2-chloropropane-1,3-diol (2-MCPD) and 3-chloropropane-1,2-diol (3-MCPD) in refined oils and fats have recently triggered an interest in the mechanism of formation and decomposition of these contaminants during oil processing. In this work, the effect of the main precursors, namely acylglycerols and chlorinated compounds, on the formation yield of MCPD esters was investigated in model systems simulating oil deodorization. The composition of the oils was modified by enzymatic hydrolysis, silica gel purification and application of various refining steps prior to deodorization (namely degumming, neutralization, bleaching). Partial acylglycerols showed greater ability, than did triacylglycerols, to form MCPD esters. However, no direct correlation was found between these two parameters, since the availability of chloride ions was the main limiting factor in the formation reaction. Polar chlorinated compounds were found to be the main chloride donors, although the presence of reactive non-polar chloride-donating species was also observed.

On-line hydrogenation of biodiesel-Toward a reference method for the determination of glycerides and total glycerol.

A reliable method with ensured traceability of the measurement results for free and bound glycerol (as monoacylglycerides, diacylglycerides and triacylglyerides) in biodiesel was developed, giving results beyond the state of the art of the current standard methodologies. The proposed method is based on an on-line hydrogenation using gas chromatography coupled to flame ionization detection and hydrogen as carrier gas. After sample introduction the hydrogenation takes place on a fused silica pre-column coated with a palladium catalyst. This approach allows an immediate and reliable hydrogenation of vegetable oils and biodiesels from different feedstocks. All glycerides are converted into their saturated analogues, resulting in simplified chromatograms with structurally clearly defined analytes, and increased sensitivity for trace amounts of compounds. The method has been successfully in-house validated and combined uncertainty values have been assigned to the final results, which were less than 8% for free glycerol, the sum of monoacylglycerides, the sum of diacylglycerides and the sum of triacylglycerides.

An experience with paediatric burn wounds treated with a plant-derived wound therapeutic.

OBJECTIVE: To observe the efficacy of a plant-derived wound dressing (1 Primary Wound Dressing®), a mixture of hypericum and neem oil, in different types of paediatric burns. METHOD: A retrospective review was conducted over the complete healing course of 9 paediatric patients with a mean age of 8.17±3.35 (1-11 years), presenting mixed, partial or full-thickness burns. The treatment applied by the wound care specialist consisted of daily cleansing of the wound with a saline solution and application of 1 Primary Wound Dressing on the whole wound surface. There was no application of a secondary dressing. The time to heal, wound size, ease of handling, pain and complications were recorded. Procedural and background pain were observed in six of the patients older than 5 years (mean age 9.6±2.39, range 8-11 years). Due to the small number of patients examined during the period studied, it was not possible to perform statistical analyses. RESULTS: The mean wound size was 50.76±48.32cm2 (4.63-132.0cm2). A rapid induction of granulation tissue and re-epithelialisation was observed. Time to complete healing was 16.6±4.69 days (10-22 days). No complications related to wound infection was observed. The 6 patients older than five years reported a strong relief of pain, from an initial value of 7-8 out of 10 to 0 out of 10 within the first week of treatment. This remained at the 0 out of 10 level during the second and third weeks of treatment. CONCLUSION: This retrospective, non-controlled examination suggests that 1 Primary Wound Dressing could be an effective therapy for the treatment of burn wounds, with benefits including pain reduction and simplicity of use. Further evaluations with a larger population are required to document the effectiveness of this plant-derived wound dressing in a controlled fashion. DECLARATION OF INTEREST: There were no external sources of funding for this study. F. Carnevali is a researcher and co-inventor of 1 Primary Wound Dressing®.

[Determination of undecanoic acid and 13-methyl-tetradecanoic acid connected to the glyceride with internal standard method and its application to the identification of adulteration of illegal cooking oil].

On the basis of the source of illegal cooking oil (heated vegetable oil and animal oil) and the important referents reflecting their sources, namely, undecanoic acid and 13-methyl-tetradecanoic acid connected to the glyceride, their corresponding ramifications in edible oil were detected with internal standard method. The sensitivity and selectivity of this method were improved by the on-line cleanup and preconcentration. The detection limits of the method were 0.070 mg/kg for undecanoic acid and 0.006 mg/kg for 13-methyl-tetradecanoic acid. Additionally, most of the normal vegetable oils have lower levels of both fatty acids than illegal cooking oils. It was suggested to evaluate the quality of edible oils to some extent on the basis of the contents of undecanoic acid and 13-methyl-tetradecanoic acid.

Green coffee oil analysis by high-resolution nuclear magnetic resonance spectroscopy.

In this work, we show how an extensive and fast quantification of the main components in green coffee oil can be achieved by NMR, with minimal sample manipulation and use of organic solvents. The approach is based on the integration of characteristic NMR signals, selected because of their similar relaxation properties and because they fall in similar spectral regions, which minimizes offset effects. Quantification of glycerides, together with their fatty acid components (oleic, linoleic, linolenic and saturated) and minor species (caffeine, cafestol, kahweol and 16-O-methylcafestol), is achieved in less than 1h making use of (1)H and (13)C spectroscopy. The compositional data obtained are in reasonable agreement with classical chromatographic analyses.