OPO结构脂在模拟婴儿胃肠道消化环境中脂肪酸的释放特性

为研究OPO（1,3-二油酸2-棕榈酸甘油三酯）结构脂在模拟婴儿胃肠道消化环境中脂肪酸释放特性,根据母乳脂肪酸组成特点,制备了OPO结构脂（O组）。以婴儿配方粉中常用植物油混合物（Z组）作对比,测定脂肪酸组成及Sn-2位脂肪酸分布情况,分析消化过程中脂肪酸释放率、粒径和zeta电位的变化,并测定了消化产物中游离脂肪酸和甘油单酯的组成。研究结果表明：相同处理条件下,O组粒径集中分布在波长198 nm处,Z组大多分布在波长229 nm处,粒径分布的不同反映在初始消化阶段脂肪酸释放率存在明显差异（P<0.05）。胃液模拟30 min后,O组脂肪酸释放率为12.83%,Z组为8.92%。经体外胃肠道消化后测得O组脂肪酸的最终释放率为62.67%,而Z组仅为51.83%。结论：O组和Z组Sn-2位脂肪酸的分布存在差异（P<0.05）,薄层色谱法验证了O组Sn-2位棕榈酸的含量较高且与母乳更接近。在模拟体外消化时表现出更高的脂肪酸释放率。消化产物甲酯化后经气-质谱分析,O组消化后产物中游离不饱和脂肪酸和棕榈酸甘油单酯含量均高于Z组。将本试验获得的OPO油脂和混合植物油脂在体外的消化脂肪酸释放特性进行比较,可得出如下结论：OPO结构脂经消化后,其脂肪酸更有利于婴儿消化吸收利用。     

人乳脂替代品1,3-二油酸-2-棕榈酸甘油三酯的研究进展

母乳是婴儿最理想的营养来源,母乳脂肪酸在甘油三酯骨架上具有高度特异性的位置分布,其70%为sn-2棕榈酸酯。棕榈酸在甘油三酯中酰基化位置的不同直接影响棕榈酸在人体内的生理功能,sn-2棕榈酸酯易被人体吸收。1,3-二油酸-2-棕榈酸甘油三酯(1,3-dioleoyl-2-palmitoyl triglyceride,OPO)是一种典型的sn-2棕榈酸酯。普通婴儿配方奶粉主要以牛乳为原料,虽然牛乳脂肪酸组成与母乳脂肪酸接近,但其脂肪酸结构与母乳存在显著差异,牛乳中的sn-2棕榈酸酯远低于母乳中sn-2棕榈酸酯含量。以牛乳为基础,在奶粉中加入1,3-二油酸-2-棕榈酸甘油三酯(OPO),较之一般的配方奶粉更加接近母乳,更宜作为人乳脂替代品?本文介绍了人乳脂替代品1,3-二油酸-2-棕榈酸甘油三酯(OPO)的研究意义、结构特点、制备方法和分析测定方法等研究进展,旨在为婴儿配方奶粉的开发提供有益参考。     

不同大分子乳化剂构建番茄红素纳米乳液的体外消化规律比较

通过体外模拟消化,研究以辛烯基琥珀酸酯化（octenyl succinic anhydride,OSA）变性淀粉、乳清分离蛋白（whey protein isolate,WPI）、酪蛋白酸钠（sodium caseinate,SC）为乳化剂构建的番茄红素纳米乳液的消化规律。结果表明,消化过程中纳米乳液的液滴大小、Zeta电位和微观结构取决于乳化剂类型,OSA变性淀粉和蛋白质类乳化剂构建的纳米乳液分别在肠和胃阶段发生水解,液滴聚集,乳液平均粒径增大,同时Zeta电位绝对值达到最小。经胃肠消化后3 种乳化剂构建的番茄红素纳米乳液游离脂肪酸释放率的大小排序为OSA变性淀粉（92.25%）＞SC（86.53%）＞WPI（79.88%）,高于对照组的48.7%,表明纳米乳液包埋体系能有效改善番茄红素的消化特性,且以OSA变性淀粉构建的纳米乳液表现出比蛋白质类乳化剂更高的番茄红素生物利用率,达到（25.60±3.08）%。     

单、双脂肪酸甘油酯(亚麻酸)乳液的制备及特性研究

针对单、双脂肪酸甘油酯（亚麻酸）极易氧化的特点,该研究以乳清浓缩蛋白（WPC）、大豆分离蛋白（SPI）、酪蛋白酸钠（SC）和吐温80（T80）为乳化剂制备乳液,考察乳化剂类型对乳液的理化性质、氧化稳定性和消化特性的影响。结果表明,乳液均具有较小的粒径（131.97~224.87 nm）,且在两周贮藏期内保持稳定。乳液包载能够提高单、双脂肪酸甘油酯（亚麻酸）的氧化稳定性,相比T80（过氧化值为377.40 mmol/kg）,蛋白质对油脂的氧化保护效果更好,其中SPI稳定的乳液过氧化值最低为197.73 mmol/kg。体外模拟消化试验表明,乳化剂类型对游离脂肪酸的释放影响较小,但蛋白稳定的乳液在胃消化阶段更容易发生液滴聚集;亚麻籽油的脂质水解程度最低为23.93%,而单、双脂肪酸甘油酯（亚麻酸）的初始消化速度更快,最终脂解程度更高（46.33%）。因此,蛋白质乳液能有效提高单、双脂肪酸甘油酯（亚麻酸）的氧化稳定性,且单、双脂肪酸甘油酯（亚麻酸）相比亚麻籽油具有更好的消化效率,有望替代亚麻籽油作为人体亚麻酸的食物来源。     

OPO乳脂粉对肠道微生物体外发酵的影响

为进一步明晰1,3-二油酸-2-棕榈酸甘油三酯（1,3-dioleoyl-2-palmitoyl triglyceride,OPO）乳脂粉的营养机制,扩大OPO的食用范围,以健康大学生粪便为菌源,采用体外厌氧发酵模式,结合传统培养技术和气相色谱法,研究不同含量OPO乳脂粉24 h体外发酵过程中pH值、短链脂肪酸的变化,并分析肠道微生物,计算益生元指数和B/E（Bifidobacterium/Enterobacter）值。结果显示：OPO乳脂粉的添加,能迅速降低发酵液的pH值,下降幅度与OPO含量呈正比;能大幅度增加总短链脂肪酸含量,其中,乙酸和丁酸含量增加明显;能显著促进双歧杆菌和乳酸杆菌的生长,抑制肠杆菌和拟杆菌的生长,提高益生元指数和B/E值,其中OPO含量50 g/kg组在24 h发酵过程中益生元指数最高,在发酵12 h内双歧杆菌增加数量、B/E值均与OPO含量呈正比。因此,综合考虑益生效率,推荐添加组为50 g/kg组。     

大豆酱油渣及低温豆粕中可溶性膳食纤维对油脂体稳定性及消化特性的影响

从大豆酱油渣及低温豆粕中提取了可溶性膳食纤维（Soybean soluble dietary fiber,SSDF）,并以Zeta电位绝对值、平均粒径、过氧化值（POV值）、硫代巴比妥酸值（TBARS值）以及游离脂肪酸（FFA）释放率等为特征参数探究了SSDF对大豆油脂体（soybean oil body,SOB）乳液的理化稳定性和消化特性的影响规律。结果表明：一定浓度的SSDF可有效降低SOB乳液体系的Zeta电位、平均粒径、POV值、TBARS值,显著改善SOB乳液的理化稳定性;此外体外模拟消化试验表明,SSDF可显著降降低SOB乳液的FFA释放率,有效延缓SOB中的脂质在胃肠道内的消化进程。本研究可为酱油渣、豆粕等加工副产物的高值化利用,为提高SOB的加工适性以及开发新型含SOB及SSDF的低脂保健食品提供理论依据及参考。     

酪蛋白凝胶颗粒对水包油型乳液稳定性及体外消化的影响

研究酪蛋白凝胶颗粒对水包油型乳液的稳定性及体外消化特性的影响,并与商业乳化剂酪蛋白酸钠和吐温20作对比。采用马尔文激光粒度仪、纳米粒度电位仪及稳定性分析仪分别测定水包油乳液的粒径、电位和稳定性;通过模拟体外消化,研究酪蛋白凝胶颗粒稳定的乳液的消化情况并比较不同种类乳化剂对乳液体外消化特性的影响。结果表明,酪蛋白凝胶颗粒稳定的乳液不稳定性指数为0.71±0.09,显著低于酪蛋白酸钠和吐温稳定乳液的不稳定性指数(分别为1.43±0.09,1.62±0.15),稳定性最好。这与酪蛋白软颗粒能以完整的颗粒结构存在于油水界面后形成较强的空间位阻和静电作用有关。体外消化结果表明,酪蛋白凝胶颗粒稳定的乳液脂肪酸的释放量为(86.13±3.91)%,而酪蛋白酸钠和吐温20稳定的乳液脂肪酸释放量分别为(86.79±3.61)%,(91.62±1.31)%,三者并没有显著性差异,表明酪蛋白凝胶颗粒对乳液消化不产生抑制作用。乳化剂种类虽对初始消化速率有影响,但不影响消化终点。结论:酪蛋白凝胶颗粒能够较好地稳定水包油乳液且不影响其消化。     

罗非鱼蛋白-乳清蛋白复合乳液负载β-胡萝卜素的研究

为了提高β-胡萝卜素的稳定性和生物利用率,充分利用双蛋白的营养和功能特性优势,以罗非鱼分离蛋白（tilapia protein isolate,TPI）和乳清分离蛋白（whey protein isolate,WPI）混合液作为乳化剂,通过高压均质结合热处理制备负载β-胡萝卜素的TPI-WPI复合乳液,探讨两种蛋白的质量比（2∶1、1∶1、1∶2）对乳液稳定性、抗氧化活性及体外消化特性的影响。结果表明,与单一的TPI和WPI乳液比较,TPI-WPI复合蛋白乳液的稳定性及β-胡萝卜素的生物利用率提高。当TPI与WPI质量比为1∶2时,复合蛋白乳液初始粒径为259.18 nm,zeta电位绝对值为28.23 mV,乳液的稳定性好,4℃贮藏21 d不分层;当TPI与WPI质量比为2∶1时,复合蛋白乳液贮藏21 d后β-胡萝卜素保留率达到62.36%,DPPH自由基和ABTS+自由基清除率分别为54.83%和40.11%,经体外消化后,β-胡萝卜素的生物利用率达24.76%,乳液游离脂肪酸释放率高。因此,WPI的添加可以提高复合蛋白乳液的稳定性,而TPI的添加可以提高乳液负载β-胡萝卜素的稳定性和生物利用率。研究结果可为混合蛋白构建稳定的乳液体系及活性成分的递送提供参考。     

新型固定化Aspergillus oryzae脂肪酶催化合成1,3-二油酸-2-棕榈酸甘油三酯

旨在评价自制固定化sn-1,3专一性脂肪酶Aspergillus oryzae L03(AOL)替代商业固定化脂肪酶酸解法合成1,3-二油酸-2-棕榈酸甘油三酯(1,3-dioleoyl-2-palmitoylglycerol,OPO)的可行性。以棕榈硬脂和油酸为原料,在正己烷体系中,得到AOL最适反应参数为:反应温度55℃,反应时间1 h,棕榈硬脂和油酸的底物摩尔比为1∶8,正己烷与底物混合物质量比1∶6,AOL水分含量3.5%(w/w)。该条件下,OPO含量44.9%,三棕榈酸甘油酯(PPP)含量为3.23%,sn-2位棕榈酸占所有棕榈酸比例为68.86%,符合国家标准。同时,AOL和LipozymeRM IM、LipozymeTL IM两种商品酶对比,AOL的催化速率最快,1 h达到最大OPO生产峰值。经过8次循环使用,AOL具有良好的操作稳定性。研究表明,自制的新型廉价固定化酶AOL具有良好的应用前景。     

1,3-二油酸-2-棕榈酸甘油三酯的合成

为了解决1,3-二油酸-2-棕榈酸甘油三酯(OPO)合成过程中不利于脂肪酶活性发挥的问题,采用两步法合成OPO,首先以棕榈硬脂和高油酸葵花籽油为原料通过化学酯交换反应得到相对棕榈硬脂熔点降低的油脂,再以化学酯交换油脂与油酸为原料在Lipozyme RM IM脂肪酶催化下进行酶法酸解反应得到OPO。通过单因素实验及响应面优化实验确定最优反应条件为棕榈硬脂与高油酸葵花籽油质量比2∶1、化学酯交换油脂与油酸质量比1∶1.45、酶添加量4%、反应温度50.40℃、反应时间5.29 h,该条件下OPO含量为27.26%,sn-2位棕榈酸占总棕榈酸含量为67.36%。以棕榈硬脂为原料合成OPO的新工艺一方面提升了高油酸植物油的使用价值,另一方面酶法酸解反应的温度适中,更有利于脂肪酶活性发挥。     

Role of calcium and calcium-binding agents on the lipase digestibility of emulsified lipids using an in vitro digestion model

Controlling the digestibility of lipids within the gastrointestinal tract is important for developing food and pharmaceutical products. In vitro digestion methods are commonly used to study the influence of formulation composition and microstructure on lipid digestibility. In this paper, we focus on the impact of calcium and calcium-binding agents on the rate of lipid droplet digestion in corn oil-in-water emulsions monitored using a pH-stat method. The rate of fatty acid production increased with increasing calcium, e.g., the free fatty acids released after 20 min digestion was <12% for 0 mM CaCl2, but >95% for 20 mM CaCl2. The ability of calcium to increase the digestion rate was found for three different emulsifiers used to stabilize the initial lipid droplets: lyso-lecithin, caseinate and β-lactoglobulin. For these three systems, the initial rate of lipid digestion increased in the following order lyso-lecithin > β-lactoglobulin > caseinate at both 0 and 20 mM CaCl2, but the rate was considerably faster at higher calcium levels for all systems. The addition of EDTA, a calcium chelating agent, to emulsions containing 20 mM CaCl2 caused an appreciable decrease in lipid digestion rate, reducing the amount of free fatty acids produced after 20 min from around 97% to 32% when the EDTA level was increased from 0 to 5 mM. Finally, we examined the impact of two anionic polysaccharides (pectin and alginate) on the rate of lipid digestion in emulsions containing 20 mM CaCl2. High methoxy pectin, which does not bind calcium strongly, did not have a major effect on the rate of digestion, whereas alginate, which does bind calcium strongly, depressed the rate considerably. This study has important implications for designing and testing delivery systems that control lipid digestion.     

The simulated in vitro infant gastrointestinal digestion of droplets covered with milk fat globule membrane polar lipids concentrate

Milk fat globule membrane polar lipids (MPL) are increasingly used as the surface-active components for emulsions in many infant food products. However, the precise effect of the emulsifier MPL on the digestion of lipids during gastrointestinal digestion has not been elucidated. This study investigated the lipid digestion of droplets covered with MPL with different sizes in a simulated in vitro infant gastrointestinal digestion assay. The well-used surface-active component casein was used as a control. Four types of emulsions were formulated: small and large droplets covered with MPL concentrate (MPL-S and MPL-L, with volumetric means of 0.35 ± 0.01 and 4.04 ± 0.01 μm, respectively), and small and large droplets covered with casein (CN-S and CN-L, with volumetric means of 0.44 ± 0.01 and 4.09 ± 0.03 μm, respectively). The emulsions were subjected to in vitro gastrointestinal digestion using a semidynamic model mimicking infant digestion. Through the determination of particle size evolution, zeta-potential, and microstructure of emulsions, the lipid droplets covered with MPL were found to be more stable than that of the CN-S and CN-L during gastrointestinal digestion. Moreover, although CN-S and CN-L showed a higher initial lipolysis rate at the beginning of gastric digestion, droplets covered by MPL exhibited a significantly higher amount of free fatty acid release during later digestion. The amount of free fatty acid release of the emulsions in both gastric and intestinal digestion could be generally classified as MPL-S ≥ MPL-L > CN-S > CN-L. Our study highlights the crucial role of MPL in the efficient digestion of emulsions and brings new insight for the design of infant food products.     

In vitro study of intestinal lipolysis using pH-stat and gas chromatography

Developing healthy products requires in-depth knowledge of digestion. This study focuses on lipid digestion in relation to emulsion properties typically followed by pH-stat. Although this is a fast and easy method to follow the overall digestion, it provides no details on lipid digestion products. Thus, the aims of the present study were to use gas chromatography (GC) to determine all products present during lipolysis, i.e. monoglycerides (MG), diglycerides (DG) and triglycerides (TG), and to compare this method with the pH-stat method for free fatty acids (FFA). Fine, medium and coarse emulsions stabilized with two different emulsifiers (whey protein isolate (WPI) or gum arabic) were digested under in vitro intestinal conditions. Although the amount of FFA increased for both methods for WPI stabilized emulsions, the amount of FFA was 2-3 times higher when determined by GC compared with pH-stat. GC analysis showed decreasing amounts of MG and DG with increasing droplet size for both emulsions. Molar ratios of FFA/DG and MG/DG were twofold higher for WPI than for gum arabic stabilized emulsions. This indicates that the total production of lipolytic products (i.e. FFA + MG + DG) depends on the droplet size and the emulsifier but their proportions only depend on the emulsifier. Although pH-stat provides a fast measure of FFA release, it is influenced by the emulsifier type at the oil-water interface and therefore care should be taken when interpreting pH-stat results. We suggest combining this method with GC for accurate FFA determination and further evaluation of all lipolytic products.     

Zein/tannic acid complex nanoparticles‐stabilised emulsion as a novel delivery system for controlled release of curcumin

The present work was aimed to evaluate the potential of the gel‐like Pickering emulsions (50%, v/v, oil) stabilised by zein/tannic acid (TA) complex colloidal particles (ZTPE) as a new encapsulation system of lipophilic ingredients. Compared with sodium caseinate‐stabilised emulsions (SCE) and bulk oil, the better protective effects of ZTPE on the chemical stability of curcumin were observed when they suffered from UV light, and the lipid oxidation rate remarkably reduced in ZTPE. Also, the zein particle layers loaded with TA around the oil droplets can provide protection against harsh gastric environment, facilitating to slow down the release of free fatty acids (FFA) and curcumin during in vitro simulated digestion. These findings show that ZTPE have a good potential to act as an efficient encapsulating agent to protect functional ingredients from degradation and control their release during digestion, which can further improve the bioavailability of bioactive ingredients.     

Effects of water-soluble soybean polysaccharide on rheological properties,stability and lipid digestibility of oil-in-water emulsion during in vitro gastrointestinal digestion

Water-soluble soybean polysaccharide (SSPS) is a naturally occurring emulsifier. SSPS was used as the sole emulsifier to stabilize an oil-in-water (O/W) emulsion. The effects were investigated of different SSPS concentrations (3–20% (w/w)) on the lipid digestibility, rheological properties and stability of O/W emulsions during in vitro digestion model. The droplet size of the emulsions tended to increase during the oral phase because the emulsions were unstable and droplets coalesced, except with a SSPS concentration of 20% (w/w). The presence of SSPS markedly reduced the free fatty acid (FFA) content after its stabilized O/W emulsion passed through in vitro gastrointestinal digestion. The amount of FFA significantly decreased as the concentration of SSPS increased due to SSPS stabilization film on oil droplet surface and high viscous system. SSPS may be an attractive alternative ingredient to control the lipid digestibility of emulsions for various food products.     

Characteristic of microencapsulated 1,3-dioleoyl-2-palmitoylglycerol and its application in infant formula powder

1,3-Dioleoyl-2-palmitoylglycerol (OPO) is used widely as a food additive. However, the structured lipid is sensitive to oxygen and light in the production process. This results in the loss of its original nutrition and production of harmful substances. In this study, the OPO was microencapsulated using whey protein isolate and maltodextrin as the wall material and monostearin as an emulsifier by spray-drying technology added to the infant formula milk, in order to improve its oxidative stability. The OPO microcapsules were released in vitro through the simulated gastrointestinal tract; sensory evaluation of microencapsulated OPO in infant formula powder and nutrition absorption of microencapsulated OPO in infant formula power by animal experiments were investigated. The results showed that OPO microcapsules have a slow-release effect; after 2 h of simulated gastric fluid digestion, only 16.1 ± 3.2% of the oil was released from microencapsulated OPO, and after another 2 h of simulated intestinal fluid digestion, there was 92.3 ± 2.8% of oil released from the microencapsulated OPO. The infant formula with microencapsulated OPO has a uniform colour and no odour. The quality of infant formula with microencapsulated OPO was obviously better than infant formula with OPO by storage test. Everted mice gut sac experiments confirmed that microencapsulation did not affect absorption of mice to OPO in infant formula and prevented the loss of calcium. The study confirmed that addition of microencapsulated OPO makes infant formula more efficient for product quality and nutrition absorption.     

Fabrication,characterization and lipase digestibility of food-grade nanoemulsions

The behavior of nanoemulsion-based delivery systems within the gastrointestinal tract determines their functional performance. In this study, the influence of particle radius (30–85 nm) on the in vitro digestion of nanoemulsions containing non-ionic surfactant stabilized lipid (corn oil) droplets was examined using simulated small intestine conditions. Nanoemulsions were prepared by a combination of high-pressure homogenization and solvent (hexane) displacement. Lipid droplets with different sizes were prepared by varying the oil-to-solvent ratio in the disperse phase prior to homogenization. The fraction of free fatty acids (FFA) released from emulsified triacylglycerols (TG) during digestion was measured by an in vitro model (pH-Stat titration). Nanoemulsions exhibited a lag-period before any FFA were released, which was explained by inhibition of lipase adsorption to the oil–water interface by free surfactant. After the lag-period, the digestion rate increased with decreasing oil droplet diameter (increasing specific surface area). The total amount of FFA released from the emulsions increased from 61% to 71% as the mean droplet radius decreased from 86 nm to 30 nm. The incomplete digestion of the emulsified lipids could be explained by inhibition of lipase activity by the release of fatty acids and/or by interactions between lipase and surfactants molecules.     

紫苏油多层乳液的制备及乳液油脂体外消化特性

针对紫苏油在贮藏过程中极易氧化的特点,以大豆分离蛋白、壳聚糖和海藻酸钠为乳化剂,采用静电层层自组装技术对紫苏油进行包封,使紫苏油保持良好的物理稳定性,并能达到油脂缓释的目的。分别对紫苏油单层乳液、双层乳液和三层乳液微观形态和稳定性进行考察,建立体外模拟消化模型,通过气相色谱测定3 种乳液消化前后的脂肪酸组成。结果表明：壁材质量分数为大豆分离蛋白1.0%、壳聚糖2.0%、海藻酸钠1.5%时制备的3 种乳液粒径较小、电位较高,具有良好的理化稳定性。在酸性条件下的多层乳液能够更好保护多不饱和脂肪酸,随着包埋层数的增加,紫苏油的氧化速率越慢。体外模拟消化结果表明三层乳液较单层和双层乳液具有更好的缓释效果,多层乳液可以保证油脂中脂肪酸有效释放。本实验阐明不同界面层对紫苏油消化和脂肪酸释放特性的影响,为指导油脂缓释体系加工提供一定的参考。