Analysis of the intestinal microbiota and profiles of blood amino acids and acylcarnitines in neonates with hyperbilirubinemia.

OBJECTIVE: This study aimed to discuss the distinctive features of the intestinal microbiota in neonates with hyperbilirubinemia and to comprehensively analyse the composition of the intestinal microbiota as well as the levels of free amino acids and acylcarnitines in the peripheral blood of neonates experiencing hyperbilirubinemia. RESULTS: At the phylum level, Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, and Chloroflexi were the five predominant microbial groups identified in both the hyperbilirubinemia and control groups. Alpha diversity analysis, encompassing seven indices, showed no statistically significant differences between the two groups. However, Beta diversity analysis revealed a significant difference in intestinal microbiota structure between the groups. Linear discriminant analysis effect size (LEfSe) indicated a significant reduction in the abundance of Gammaproteobacteria and Enterobacteriaceae within the hyperbilirubinemia group compared to that in the control group. The heatmap revealed that the control group exhibited increased abundances of Escherichia and Bifidobacterium, while the hyperbilirubinemia group exhibited increased levels of Enterococcus and Streptococcus. Regarding blood amino acids and acylcarnitines, there were greater concentrations of citrulline (Cit), arginine (Arg), ornithine (Orn), and valine (Val) in the hyperbilirubinemia group than in the control group. The hyperbilirubinemia group also exhibited significant increases in medium-chain fatty acids (C6, C8), long-chain fatty acids (C18), and free carnitine (C0). CONCLUSION: By comparing neonates with hyperbilirubinemia to those without, a significant disparity in the community structure of the intestinal microbiota was observed. The intestinal microbiota plays a crucial role in the bilirubin metabolism process. The intestinal microbiota of neonates with hyperbilirubinemia exhibited a certain degree of dysbiosis. The abundances of Bacteroides and Bifidobacterium were negatively correlated with the bilirubin concentration. Therefore, the fact that neonates with hyperbilirubinemia exhibit some variations in blood amino acid and acylcarnitine levels may provide, to a certain degree, a theoretical basis for clinical treatment and diagnosis.

Evaluation of high oleic sunflower oil oleogels with beeswax, beeswax-glyceryl monopalmitate, and beeswax-Span80 in cookie preparation.

BACKGROUND: Shortening is used widely in cookie preparation to improve quality and texture. However, large amounts of saturated and trans fatty acids present in shortening have adverse effects on human health, and much effort has been made to reduce the use of shortening. The use of oleogels might be a suitable alternative. In this study, the oleogels of high oleic sunflower oil with beeswax (BW), BW-glyceryl monopalmitate (BW-GMP), and BW-Span80 (BW-S80) were prepared and their suitability to replace shortening in cookie preparation was evaluated. RESULTS: The solid fat content of BW, BW-GMP, and BW-S80 oleogels was significantly lower than that of commercial shortening when the temperature was not higher than 35 °C. However, the oil-binding capacity of these oleogels was almost similar to that of shortening. The crystals in the shortening and oleogels were ß' form mainly; however, the morphology of crystal aggregates in these oleogels was different from that of shortening. The textural and rheological properties of doughs prepared with the oleogels were similar, and clearly different from those of dough with commercial shortening. The breaking strengths of cookies made with oleogels were lower than that of cookies prepared with shortening. However, cookies containing BW-GMP and BW-S80 oleogels were similar in density and color to those prepared with shortening. CONCLUSION: The textural properties and color of cookies with BW-GMP and BW-S80 oleogels were very similar to those of the cookies containing commercial shortening. The BW-GMP and BW-S80 oleogels could act as alternatives to shortening in the preparation of cookies. © 2023 Society of Chemical Industry.

Enhanced transesterification of ethyl ferulate with glycerol for preparing glyceryl diferulate using a lipase in ionic liquids as reaction medium.

Glyceryl diferulate (DFG) is a water-soluble ester of ferulic acid. A novel ionic liquid (IL) system for enzymatic transesterification of ethyl ferulate (EF) with glycerol to produce DFG was developed. Of three ILs with different anions (BF4 (-), PF6 (-) and TF2N(-)) and cations (BDMIM, OMIM, HMIM, BMIM, and EMIM), EMIMTF2N proved the best using a commercial lipase. It had a significant protective effect against thermal inactivation of the enzyme. High EF conversion (~100 %) and DFG yield (45 %) were achieved using 45 mg enzyme/ml; temperature, 70 °C; reaction time, 12 h.

Effects of food formulation on bioavailability of phytosterols: phytosterol structures, delivery carriers, and food matrices.

Daily intake of phytosterols (PSs) as a diet supplement can lower blood-cholesterol levels and reduce the risks of cardiovascular diseases. However, the high crystallinity, low water solubility, easy oxidizability, and other characteristics of PSs restrict their application and bioavailability in food products. The formulation parameters including the structures of PSs, delivery carriers, and food matrices may play an important role in the release, dissolution, transport, and absorption of PSs in functional foods. In this paper, the effects of formulation parameters, including phytosterol structures, delivery carriers, and food matrices, on the bioavailability of phytosterols are summarized and suggestions are provided for the formulation design of functional foods. The side chain and hydroxyl esterification group of PSs may significantly affect their lipid or water solubilities and micellization capacities, which in turn affect the bioavailability of PSs. Selecting suitable delivery carriers based on the characteristics of the food system can reduce the crystallinity and oxidation of PSs and control the release of PSs, thereby improving the PS stability and delivery efficiency. Moreover, the ingredients of the carriers or food products would also influence the release, solubility, transport, and absorption of PSs in the gastrointestinal tract (GIT).

Deep eutectic solvents as switchable solvents for highly efficient liquid-liquid microextraction of phenolic antioxidant: Easily tracking the original TBHQ in edible oils.

Synthetic antioxidant tert-butylhydroquinone (TBHQ) is easily oxidized to tert-butylquinone (TQ) during the storage of edible oils, resulting in an obvious decrease in the content of TBHQ in edible oils. Therefore, it is quite desirable to develop a simple analytical method for accurately tracking the original content of TBHQ in edible oils. In this work, deep eutectic solvents (DESs) have been successfully used in room temperature vortex-assisted liquid-liquid microextraction (VALLME) of TBHQ from edible oils. The DES composed of ethylene glycol and choline chloride (ChCl) could selectively extract TBHQ from edible oils containing both TBHQ and TQ. The DES composed of sesamol and ChCl (molar ratio of 3:1) could efficiently reduce TQ to TBHQ and extract TBHQ from edible oils. The whole VALLME process only required 5 min at room temperature. This switchable DESs-based VALLME with common RP-HPLC analysis showed high efficiency and good performance with linearity (R2 = 0.9999) in 5-500 mg/kg range, detection limit of 0.02 mg/kg, recoveries of 96.1-106.0% and intra-/inter-day precision below 2.0%. This analytical method is suitable for detecting the current content of TBHQ and tracking the original content of TBHQ during the storage of edible oils at room temperature, respectively.

Thermal Properties, Microstructure and Crystallization of Blends of Leaf Lard and Cottonseed Oil Stearin.

Blend oils composed by leaf lard (LL) and cottonseed oil stearin (COS) were prepared and the thermal property, microstructure and crystallization of these blends were investigated in the present study. Solid fat content (SFC), thermal behaviors, triacylglycerols composition, crystal structure and morphology of the LL and COS blends were determined by pulsed nuclear magnetic resonance (pNMR), differential scanning calorimetry (DSC), high-performance liquid chromatography (HPLC), X-ray diffractometer (XRD) and polarized light microscope (PLM), respectively. SFC profiles and iso-solid diagrams indicated that SFCs of all blends were almost close to the weighted averages of the fat components at temperatures beyond 20°C; however, below 20°C, SFCs of blends exhibited higher than those of the weighted averages of the fat components. With the content of COS increasing, palmitic acid and linoleic acid in the blends increased, while stearic acid and oleic acid decreased; monounsaturated-disaturated (USS) and triunsaturated (UUU) glycerides in the blends enhanced, while monosaturated-diunsaturated (UUS) glycerides declined. The melting temperature of the blends decreased with the increase of COS content. The crystal forms in LL were ß' and ß, and the packing pattern was double and triple chain length (2L and 3L). With COS in blends increasing, ß' form crystals and 3L pattern reduced. Polarized light micrographs showed that the number of crystal particles in the blends raised with the increase of COS content, meanwhile, the grainsize of the sample gradually decreased. Visual appearances of the blends indicated that blending LL with COS could efficiently reduce the graininess of LL. The addition of COS had a significant effect on the crystallization behavior of LL. LL presented one-step crystallization at 10°C and 20°C, while COS showed two-step crystallization at 10°C and one-step crystallization at 20°C. However, the blends exhibited obvious two-step crystallization at 10°C, one-step or slight two-step crystallization at 20°C.

Solvent-free synthesis of phytosterol linoleic acid esters at low temperature.

Phytosterol unsaturated fatty acid esters show much higher oil solubility than free phytosterol. Thus, development of a green and low-cost method for the preparation of phytosterol fatty acid esters is highly desirable in the food industry. Herein, we have developed a simple chemical method toward efficient preparation of phytosterol linoleic acid esters at very mild temperature (60 °C) using 4-dodecylbenzenesulfonic acid (DBSA) as the catalyst. In this work, low-temperature esterification of phytosterols (soybean sterol) with linoleic acid could produce the corresponding phytosterol esters above 95% conversion under solvent-free conditions. In addition, this simple method could be applied to produce phytosterol esters through esterification of phytosterol with an unsaturated fatty acid mixture resulting from the hydrolysis of various vegetable oils. Importantly, no extra organic solvents and no extra water-removal operations or equipment were required in this chemical esterification method. The mechanism investigation suggested that the DBSA-catalyzed low-temperature esterification would form micro-emulsions of water-in-oil (W/O), which could achieve automatic separation of water from the hydrophobic system to avoid reverse reaction hydrolysis and rapidly promote the equilibrium reaction towards phytosterol esters.

Enzymatic preparation of phytosterol esters with fatty acids from high-oleic sunflower seed oil using response surface methodology.

Phytosterol esters are functional compounds that can effectively reduce plasma cholesterol concentration, and have wide applications in the food industry. In this study, a simple and efficient enzymatic method was successfully applied to synthesize phytosterol oleic acid esters with fatty acids from high-oleic sunflower seed oil. Among the tested lipases, Candida rugosa lipase (CRL) exhibited higher catalytic activity in the esterification of phytosterols with fatty acids (oleic acid 84%) from high-oleic sunflower seed oil. Box-Behnken design and response surface methodology were used to investigate the influence of reaction factors on the conversion of phytosterols. The maximum conversion of phytosterols (96.8%) and yield of phytosterol esters (92%) could be obtained under optimal conditions: reaction temperature 50 °C, a molar ratio of phytosterols to fatty acids at 1 : 2.3, enzyme loading of 5.8%, isooctane volume of 2 mL and reaction time of 2 h. It was noteworthy that this enzymatic esterification method indeed expended a much shorter reaction time (2 h) than that observed in previous reports. In general, the enzymatic preparation of phytosterol oleic acid esters with fatty acids from high-oleic sunflower seed oil will be a simple and rapid method for producing unsaturated fatty acid esters of phytosterol with both higher oil solubility and oxidative stability, which is beneficial as functional food ingredients.

Chemoenzymatic synthesis of feruloylated monoacyl- and diacyl-glycerols in ionic liquids.

Feruloylated monoacyl- and diacyl-glycerols (FMAGs and FDAGs) are lipophilic antioxidants and potential UV absorbers. FMAGs and FDAGs were synthesized by a novel chemoenzymatic method: firstly, ferulic acid was esterified with glycerol to synthesize glyceryl ferulate, using p-toluenesulfonic acid as chemical catalyst in 1-butyl-3-methylimidazolium tetrafluoroborate ([Bmim]BF(4)); secondly, glyceryl ferulate was esterified with oleic acid to synthesize FMAGs and FDAGs, using Novozym 435 as biocatalyst in 1-butyl-3-methylimidazolium hexafluorophosphate ([Bmim]PF(6)). The conversion of ferulic acid and yield of glyceryl ferulate in the first reaction were both 98%. The yields of FMAGs and FDAGs in the second reaction reached 34 +/- 2% and 66 +/- 3%, respectively.

A highly efficient microextraction technique based on deep eutectic solvent formed by choline chloride and p-cresol for simultaneous determination of lignans in sesame oils.

For simultaneous determination of lignans (i.e. sesamin, sesamolin and sesamol) in sesame oils, a highly efficient ultrasonic-assisted liquid-liquid microextraction (UALLME) technique was developed based on deep eutectic solvents (DESs) composed of choline chloride and p-cresol. Compared with the common polyols-based DESs (i.e. choline chloride-ethylene glycol), the phenolic DESs exhibited higher extraction efficiency for both polar lignans (i.e. sesamol) and non-polar lignans (i.e. sesamin and sesamolin) due to the π-π interaction between lignans and DES. After the UALLME process, DES phase was analyzed by HPLC-UV. Under optimized condition, all three lignans showed good linear calibration curves and low limit of detection. The recoveries were in the range of 97.3-120%, and the intra-day and inter-day precisions were less than 0.97%. Moreover, this method was used for simultaneous determination of lignans in 15 commercial oil samples. The results showed that the new microextraction technique achieved similar efficiency as the conventional liquid-liquid extraction method.

Amano Lipase PS-catalyzed Hydrolysis of Pine Nut Oil for the Fatty Acids Production Using Deep Eutectic Solvent as Co-solvent.

Free fatty acids (FFAs) are the important material used in food, personal care, emulsifiers, adhesives and surfactants. In order to enhance the preparation of FFAs, the effects of reaction variables, optimization, thermodynamic property for the Amano lipase PS catalyzed hydrolysis of pine nut oil (PNO) using deep eutectic solvents (DESs) as co-solvents were studied. The results showed that FFAs could be successfully prepared from pine nut oil through Amano lipase PS catalyzed hydrolysis using Choline chloride:Urea (ChCl:U, 1:2, mol/mol) as co-solvent. Under the optimal conditions (reaction temperature 46°C, water amount 38%, DES addition 43%, lipase dosage 7.6%, reaction time 13 h), the maximum content of FFAs in the products and degree of hydrolysis (DH) of oil were up to 89.1 ± 1.9% and 92.7 ± 2.2%, respectively. The effects of reaction variables on the hydrolysis increased in the order of DES addition < reaction temperature < reaction time < lipase dosage < water amount. The thermodynamics (Arrhenius equation) for the triglycerides hydrolysis was V = 4289.39·exp(-22942.09/RT) with the activation energy (Ea) of 22.94 kJ/mol. The Gibbs free energy (ΔG), enthalpy (ΔH) and entropy (ΔS) were 81.50 ± 2.64 kJ/mol, 20.18 ± 0.12 kJ/mol and -184.59 ± 0.36 J/mol/K, respectively. The lipase in the aqueous DES could be directly re-used for 3 times.

Engineering phytosterol-based oleogels for potential application as sustainable petrolatum replacement.

Phytosterol-based oleogels have been engineered in edible oils for potential applications as sustainable replacements for petrolatum. Oleogels have emerged with a crystal network structure with oil molecules trapped inside. In addition, the viscosity of highly thixotropic oleogels could be tuned by manipulating the concentration of phytosterols and monoglycerides, and the type of surface-active small molecules and bulk vegetable oils. Furthermore, viscous soft matter could also be tunably made with 8-20% oleogelators in olive oil with favourable water vapour occlusive and wettability properties, in addition to having good texture, and outstanding thixotropic and thermal reversibility properties. These properties are quite similar to those of commercial petrolatum. This work demonstrates that the natural phytosterol-oleogels in edible oils can be a novel source of sustainable and green replacements for petrolatum.

Characterization of the chemical composition of lotus plumule oil.

Characterizations of lotus plumule and plumule oil, focusing on approximate composition analysis of lotus plumule powder and fatty acid composition, lipid classes, triglyceride (TG) profiles, and sterol analysis of the plumule oil, were conducted in this work. The results revealed that the lotus plumule constitutes 7.8% moisture, 4.2% ash, and 12.5% crude oil and 26.3% protein on the dry base. Lotus plumule oil is rich in linoleic acid (50.4%) and oleic acid (13.5%), and the dominating saturated fatty acids are palmitic acid (18.0%) and behenic acid (6.8%). The principal components of TG in lotus plumule oil are LLL (12.80%), beta-PLL (11.27%), beta-POL (8.28%), beta-PLO (8.58%), and beta-BeLL (8.32%). Lipid class assay of the crude oil gave the saponification value of 153.4 KOH mg/g and tocopherol content 390 mg/100 g. A distinct characteristic of lotus plumule oil is that its unsaponifiable matter is incredibly high, up to 14-19%, which consists mainly of beta-sitosterol (32%), Delta(5)-avenasterol (20%), and campesterol (6.3%). The major occurring form of sterols was found to be steryl ester. This work might be useful to develop innovative applications of lotus plumule oil.

Thermal Losses of Tertiary Butylhydroquinone (TBHQ) and Its Effect on the Qualities of Palm Oil.

The rules and patterns of thermal losses of tertiary butylhydroquinone (TBHQ) in palm oil (PO) and its effect on the qualities of PO were investigated by oven heating method. Volatilization and transformation products of TBHQ in PO were also studied in detail under heating treatment. Results showed that at low temperature (< 135°C), TBHQ had better antioxidative properties, while its antioxidative potency to PO was significantly weakened at high temperature (≥ 135°C). In addition, as heating temperatures increased and heating time prolonged, losses of TBHQ significantly increased in PO. Volatilization was the major pathway for losses of TBHQ in PO under heating treatment. Meanwhile, a small portion of TBHQ was transformed and the major transformation product was 2-tertbutyl-1,4- benzoquinone (TQ). Moreover, TQ and several decomposition products of PO were also observed in the volatilization products of TBHQ.

Increase of Oleic Acid Content in Phosphatidylcholine through Lipase-catalyzed Interesterification: Optimization by Response Surface Methodology.

In order to obtain phosphatidylcholine (PC) with higher amount of oleic acid, the interesterification between soybean PC and Camellia oleifera oil (COO) rich in oleic acid catalyzed by lipase was studied in hexane. For this aim three commercially available immobilized lipases (Novozym 435, Lipozyme TLIM and Lipozyme RMIM) were assayed and Novozym 435 was finally selected for further optimization. The effects of the factors, such as PC concentration, substrate ratio, water amount, lipase dosage and temperature, on the oleic acid content in PC and PC recovery during the interesterification were investigated. The conditions of the interesterification were optimized using response surface methodology. The optimum conditions were as follows: lipase dosage 13 % (based on the mass of PC and COO), reaction temperature 55°C, water amount 5% (based on the mass of PC), reaction time 8 h, PC concentration 0.3g/mL (PC/hexane), PC-to-COO ratio 1:3 (acyl groups in PC/acyl groups in COO, mol/mol). Under these conditions, oleic acid content and PC recovery were 40.8 ± 0.5% and 69.0 ± 2.8%, respectively. Analysis of variance (ANOVA) showed that the regression models were adequate for predicting the interesterifiction. The orders of reaction variables affecting on oleic acid content and PC recovery were water amount > reaction time > lipase dosage > reaction temperature, and water amount > reaction temperature > lipase dosage > reaction time, respectively.

Lysophosphatidylcholine synthesis by lipase-catalyzed ethanolysis.

Lysophosphatidylcholine (LPC) is amphiphilic substance, and possesses excellent physiological functions. In this study, LPC was prepared through ethanolysis of phosphatidylcholine (PC) in n-hexane or solvent free media catalyzed by Novozym 435 (from Candida antarctica), Lipozyme TLIM (from Thermomcyces lanuginosus) and Lipozyme RMIM (from Rhizomucor miehei). The results showed that three immobilized lipases from Candida Antarctica, Thermomcyces lanuginosus and Rhizomucor miehei could catalyze ethanolysis of PC efficiently. In n-hexane, the LPC conversions of ethanolysis of PC catalyzed by Novozyme 435, Lipozyme TLIM and Lipozyme RMIM could reach to 98.5 ± 1.6%, 94.6 ± 1.4% and 93.7 ± 1.8%, respectively. In solvent free media, the highest LPC conversions of ethanolysis of PC catalyzed by Novozyme 435, Lipozyme TL IM and Lipozyme RM IM were 97.7 ± 1.7%, 93.5 ± 1.2% and 93.8 ± 1.9%, respectively. The catalytic efficiencies of the three lipases were in the order of Novozyme 435 > Lipozyme TLIM > Lipozyme RMIM. Furthermore, their catalytic efficiencies in n-hexane were better than those in solvent free media.

Detection and quantification of adulteration of sesame oils with vegetable oils using gas chromatography and multivariate data analysis.

This study was performed to develop a hierarchical approach for detection and quantification of adulteration of sesame oil with vegetable oils using gas chromatography (GC). At first, a model was constructed to discriminate the difference between authentic sesame oils and adulterated sesame oils using support vector machine (SVM) algorithm. Then, another SVM-based model is developed to identify the type of adulterant in the mixed oil. At last, prediction models for sesame oil were built for each kind of oil using partial least square method. To validate this approach, 746 samples were prepared by mixing authentic sesame oils with five types of vegetable oil. The prediction results show that the detection limit for authentication is as low as 5% in mixing ratio and the root-mean-square errors for prediction range from 1.19% to 4.29%, meaning that this approach is a valuable tool to detect and quantify the adulteration of sesame oil.

Solvent-free enzymatic transesterification of ethyl ferulate and monostearin: optimized by response surface methodology.

In this study, enzymatic transesterification of ethyl ferulate (EF) and monostearin for feruloylated lipids production was investigated. Enzyme screening and the effect of feruloyl acceptors on the transesterification were also studied. Effects of reaction variables (reaction temperatures, enzyme load, and reaction time) on the transesterification were optimized using response surface methodology (RSM). The optimum conditions were as follows: reaction temperature 74°C, reaction time 23h, and enzyme load 20% (w/w, relative to the weight of substrates). Under these conditions, EF conversion was 98.3±1.1%, and the transesterification product was consisted of 19.2±2.1% glyceryl ferulate (FG), 32.9±1.9% diferuloylated glycerols (DFG), 36.6±2.2% feruloylated monoacylglycerols (FMAG), 9.1±2.0% feruloylated diacylglycerols (FDAG), and 0.5% ferulic acid (FA). Analysis of variance (ANOVA) showed that the regression equation was adequate for predicting EF conversion. The activation energies for hydrolysis to form FG+DFG and transesterification to form FMAG+FDAG were calculated as 22.45 and 51.05kJ/mol, respectively, based on Arrhenius law.