Xiaoqiang Zou

Human Milk Fat Globules from Different Stages of Lactation: A Lipid Composition Analysis and Microstructure Characterization

The physicochemical properties of human milk fat globules (MFG) at different lactation stages from Danish mothers and the microstructure changes of MFG membrane (MFGM) at varied temperatures were investigated, and the relationship between chemical composition and the microstructure of MFGM was elucidated. The fat content in MFG was found to be significantly increased as lactation progressed, and colostrum MFG had the largest mean diameter of 5.75 ± 0.81 μm and the lowest ζ potential of -5.60 ± 0.12 mV. Chemical composition analyses of MFG revealed the following: (i) Colostrum milk fat constituted higher content in PUFAs (ω-6, and long-chain ω-6 and ω-3) than transitional and mature milk fats, with the corresponding lower content of SFA in its sn-2 position. (ii) The content of polar lipids among total lipids varied during lactation course (maximized at transitional stage); however, in terms of subclasses of polar lipids, no significant change of the relative content of sphingomyelin was observed, while the content of phosphatidycholine in mature milk was higher than that in colostrum and transitional milk. (iii) Inspection of fatty acid composition in phospholipids from different lactation milk revealed no remarkable and regular changes could be generalized; and no obvious difference of the morphologies of MFGM at different lactation stages can be visualized. An investigation of the microstructure change of MFGM vs temperature demonstrated that the segregated domains became larger as temperature decreased to 4 °C, while it became smaller when increased to 37 °C. This phenomenon indicated that, in addition to sphingimyelin and cholesterol, phospholipids might also contribute to increasing the segregated domains at lower temperature, while, at elevated temperature, these domains could be diminished, most likely due to a restructuring or distributing of sphingimyelin and cholesterol.

Lipid Composition Analysis of Milk Fats from Different Mammalian Species: Potential for Use as Human Milk Fat Substitutes

The lipid compositions of commercial milks from cow, buffalo, donkey, sheep, and camel were compared with that of human milk fat (HMF) based on total and sn-2 fatty acid, triacylglycerol (TAG), phospholipid, and phospholipid fatty acid compositions and melting and crystallization profiles, and their degrees of similarity were digitized and differentiated by an evaluation model. The results showed that these milk fats had high degrees of similarity to HMF in total fatty acid composition. However, the degrees of similarity in other chemical aspects were low, indicating that these milk fats did not meet the requirements of human milk fat substitutes (HMFSs). However, an economically feasible solution to make these milks useful as raw materials for infant formula production could be to modify these fats, and a possible method is blending of polyunsaturated fatty acids (PUFA) and 1,3-dioleoyl-2-palmitoylglycerol (OPO) enriched fats and minor lipids based on the corresponding chemical compositions of HMF.

Lipase-catalyzed preparation of human milk fat substitutes from palm stearin in a solvent-free system.

Human milk fat substitutes (HMFSs) were synthesized by lipozyme RM IM-catalyzed acidolysis of chemically interesterified palm stearin (mp = 58 °C) with mixed FAs from rapeseed oil, sunflower oil, palm kernel oil, stearic acid, and myristic acid in a solvent-free system. Response surface methodology (RSM) was used to model and optimize the reactions, and the factors chosen were reaction time, temperature, substrate molar ratio, and enzyme load. The optimal conditions generated from the models were as follows: reaction time, 3.4 h; temperature, 57 °C; substrate molar ratio, 14.6 mol/mol; and enzyme load, 10.7 wt % (by the weight of total substrates). Under these conditions, the contents of palmitic acid (PA) and PA at sn-2 position (sn-2 PA) were 29.7 and 62.8%, respectively, and other observed FAs were all within the range of FAs of HMF. The product was evaluated by the cited model, and a high score (85.8) was obtained, which indicated a high degree of similarity of the product to HMF.

Model for Human Milk Fat Substitute Evaluation Based on Triacylglycerol Composition Profile

Being the dominant components in human milk fat (HMF), triacylglycerol (TAG) composition might be the best approximation index to represent the composing characteristics of HMF. In this study, TAG composition of HMF from different lactation stages was analyzed by RP-HPLC-APCI-MS, and the establishment of a model for the precise evaluation of human milk fat substitutes (HMFSs) based on TAG composition was indirectly realized by employment of fatty acid composition and distribution and polyunsaturated fatty acid (PUFA) and TAG compositions. The model was verified by the selected fats and oils with specific chemical compositions, and the results revealed the degrees of similarity of these fats and oils in different evaluation aspects reflected their differences in corresponding chemical composition with HMF. The newly established evaluation model with TAG composition as a comparison base could provide a more accurate method to evaluate HMFSs and might have some inspirations for HMFS production in the future.

Identification of phospholipids classes and molecular species in different types of egg yolk by using UPLC-Q-TOF-MS

Egg phospholipids (PLs) are currently the products of greatest commercial interest with major area of importance in various fields. Therefore, in this study, duck, hen and quail egg yolk PLs were isolated by solvent extraction with chilled acetone precipitation, and subsequently separated and identified by using ultra-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Egg PLs were separated on hydrophilic interaction liquid chromatography (HILIC) with ethylene bridged hybrid (BEH) column by gradient elution using acetonitrile/ammonium formate as a mobile phase, and detected by mass spectrometry (MS) under electrospray ionization in positive and negative ion mode. Structural characterizations of 57 molecular species of egg yolk PLs were identified based on MS/MS fragment ion information and elemental composition in MassLynx 4.1 software. The obtained results showed that phosphatidylcholine (16:0-18:1), phosphatidylethanolamine (18:0-20:4), phosphatidylinositol (18:0-18:2), phosphatidylserine (18:0-18:2), sphingomyelin (d18:1/16:0) and lysophosphatidylcholine (16:0) were the predominant species among the different classes of egg yolk phospholipids.

Influence of ionic liquids on lipase activity and stability in alcoholysis reactions

Lipase activity and stability in ionic liquids containing N,N-dialkylimidazolium cations and different anions were investigated in alcoholysis reactions. Anions were the main factor affecting enzyme activity and stability, while hydrophobic and hydrophilic ionic liquids showed quite different phenomena. The initial enzyme activity in [Tf2N]− and [PF6]− based ionic liquids was much higher than that in ionic liquids containing anions such as [BF4]−, [N(CN)2]− and [OAc]−. Novozyme 435 retained approximately 5%, 40% and 100% of the blank sample activity after being immersed in [BF4]−, [PF6]− and [Tf2N]− based ionic liquids for 24 h. To reveal the influence of the ionic liquid properties, the initial enzyme activities were correlated with hydrophobicity (log P), polarity (ENT), hydrogen bond basicity (β) and viscosity. The initial enzyme activity increased with log P value and decreased with ENT value. In addition, the initial enzyme activity increased with β value in a narrow range (0.24–0.26) and then decreased continuously with the increase of β value. For [Tf2N]− based ionic liquids, the initial enzyme activity increased with viscosity, while the adverse relationship was found in [BF4]− and [PF6]− based ionic liquids. The enzyme secondary structure changes in ionic liquids were also investigated by FT-IR, which showed that the α-helix content decreased and the β-sheet content increased with the increase of immersion time.

β-d-Glucosidase-Catalyzed Deglucosidation of Phenylpropanoid Amides of 5-Hydroxytryptamine Glucoside in Safflower Seed Extracts Optimized by Response Surface Methodology

Beta-D-glucosidase-catalyzed deglucosidation of phenylpropanoid amides of 5-hydroxytryptamine (PAHAs) glucoside in safflower (Carthamus tinctorius L.) seed extracts, including N-(p-coumaroyl)serotonin glucoside (CSG) and N-feruloylserotonin glucoside (FSG), was optimized by response surface methodology (RSM). The Box-Behken design (BBD) was employed to evaluate the interactive effects of independent variables on the deglucosidation rates of CSG and FSG. The variables involved were pH (5.6-6.2), temperature (45-55 degrees C), and enzyme load (2.0-3.0%, relative to the weight of the total substrate). The substrate concentration was fixed at 3.3 g/L on the basis of factorial experiments. The optimum conditions obtained via RSM at a fixed time of 2 h were as follows: pH, 5.9; temperature, 48 degrees C; and enzyme load, 3.0%. Under these conditions, the actual deglucosidation rates of CSG and FSG were 75.5 and 42.2%, respectively, which agree well with the predicted values (75.3 and 41.9%) by RSM. The final incubation time (10 h) was determined by the time course of the deglucosidation under the above-mentioned optimum conditions, which gave the deglucosidation rates of both CSG and FSG above 90%. Simultaneously, 2-hydroxyarctiin, a typical cathartic beta-glucoside, was also removed by 80.3%.

Microstructural and lipid composition changes in milk fat globules during milk powder manufacture

The purpose of this study was to investigate the effects of milk powder processing conditions (pasteurisation, homogenisation and spray-drying) on the microstructure and composition of fat globules in cow milk. In general, the process of pasteurisation results in fewer changes in the microstructure and lipid composition than do other processing methods. The phospholipid, sterol, and the fatty acid composition of the phospholipids, with the exception of the fatty acids of total lipids, underwent significant changes under different processing conditions compared with the untreated milk fat globules. The contents and distribution of the phospholipids in pasteurised milk indicated a high level of stability, but the amounts of phosphatidylethanolamine, phosphatidylserine, and sphingomyelin were significantly affected by homogenisation. A reduction in the cholesterol content was observed after pasteurisation and homogenisation. The results indicate that processing had a significant effect on the composition and structure of the membrane of the phospholipids in milk fat globules.

Combined urea-thin layer chromatography and silver nitrate-thin layer chromatography for micro separation and determination of hard-to-detect branched chain fatty acids in natural lipids

A simple, fast and efficient procedure was developed for micro separation and enrichment of branched chain fatty acids (BCFA) from natural products using successive thin layer chromatography (TLC) technique coupling novel urea-TLC with AgNO3-TLC, which rely on the formation of urea adduction and AgNO3 bonding in methanol. These natural lipids contain a significant amount of straight chain fatty acids (FA). Fresh and fast urea-TLC and AgNO3-TLC plate making techniques were developed with more even coating and less coating material contamination before being utilized for separation. Goat milk fat was used as a model. Various experimental parameters that affect urea-TLC and AgNO3-TLC separation of BCFA were investigated and optimized, including coating of urea, concentration of original oil sample, mobile phase and sample application format. High efficiency of removal of straight chain FA was achieved with a low amount of sample in an easy and fast way. A total BCFA mix with much higher purity than previous studies was successfully achieved. The developed method has also been applied for the concentration and analysis of BCFA in cow milk fat and Anchovy oil.

Synthesis of 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids by lipase-catalyzed acidolysis of microbial oil from Mortierella alpina

Microbial oils (MOs) have gained widespread attention due to their functional lipids and health promoting properties. In this study, 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids (SLs) were produced from MO and oleic acid (OA) via solvent-free acidolysis catalyzed by Lipozyme RM IM. Under the optimal conditions, the content of unsaturated fatty acids (UFAs) increased from 60.63 to 84.00%, while the saturated fatty acids (SFAs) content decreased from 39.37 to 16.00% at sn-1,3 positions in SLs. Compared with MO, arachidonic acid (ARA) content at the sn-2 position of SLs accounted for 49.71%, whereas OA was predominantly located at sn-1,3 positions (47.05%). Meanwhile, the most abundant triacylglycerol (TAG) species in SLs were (18:1-20:4-18:1), (20:4-20:4-18:1), (18:1-18:2-18:1), (18:1-18:2-18:0) and (24:0-20:4-18:1) with a relative content of 18.79%, 11.94%, 6.07%, 5.75% and 4.84%, respectively. Such novel SLs with improved functional properties enriched with UFAs are highly desirable and have the potential to be used in infant formula.