1,3-甘油二酯的制备研究进展

1,3-甘油二酯是一种重要的功能油脂,而且是一种重要的药物中间体,对其制备具有重要意义。基于此,从1,3-甘油二酯的分析测定、合成方法到分离提取来制备1,3-甘油二酯的一整套过程的研究进展进行了综述。1,3-甘油二酯的分析测定方法主要有高效液相色谱法、气相色谱法和薄层层析法;其合成方法主要有化学法和生物法,其中生物法绿色健康、环保友好,是一种很有发展前景的方法;分离提取目前主要有层析法、分子蒸馏法和溶剂结晶法,新的分离方法还有待去探索开发如分子印迹法等。     

甘油二酯对可可脂类油脂结晶和巧克力起霜的影响研究进展

可可脂类油脂富含对称型单不饱和甘油三酯,通过科学调控可可脂类油脂中的甘油二酯,可优化巧克力的调温工艺,控制其起霜行为。综述了可可脂类油脂中甘油二酯的含量与种类(包括脂肪酸饱和度、链长和位置分布),探讨其对油脂结晶行为的作用规律,进而分析其对巧克力抗霜性能的影响。在适宜含量下1,3-甘油二酯可促进油脂形成巧克力需要的Ⅴ型晶体结构,而1,2-甘油二酯则倾向于使油脂形成Ⅱ～Ⅳ型晶体结构,且作用程度受脂肪酸链长的影响。然而,高含量甘油二酯无法使油脂形成Ⅴ型晶体结构。当甘油二酯含量在3%～5%时,油脂虽可形成Ⅴ型晶体结构,但需根据具体含量对结晶参数进行调整。因此,有必要通过分提、复配等技术对可可脂类油脂的甘油二酯进行调控。     

水活度对全细胞脂肪酶GZUF36催化合成1,3-甘油二酯中的影响

1,3-甘油二酯是一种健康油脂,并且可用来合成药物的中间材料,但其在天然油脂中含量有限。所以本文用黑曲霉GZUF36全细胞脂肪酶法甘油解合成了1,3-甘油二酯,重点探讨了影响其合成的关键因素:水活度。控制水活度的方法包括用饱和盐溶液平衡酶粉、水合盐平衡反应体介质、水合盐控制水活度水合盐分别预平衡酶粉和反应介质。结果表明,水活度显著地影响1,3-甘油二酯的产量和脂肪酶对1,3-甘油二酯的选择性。过高或过低的水活度都不利于全细胞脂肪酶GZUF36催化甘油解反应合成1,3-甘油二酯,水活度过低会导致酶活力不高,水活度过高则易使催化反应向水解方向进行。当用aw=0.58的水合盐分别预平衡酶粉和反应介质时,1,3-甘油二酯的得率和脂肪酶对1.3-甘油二酯的选择性都为最优,分别是26.100%和83.056%。本文为进一步优化全细胞脂肪酶GZUF36催化的选择性合成1,3-甘油二酯中的反应体系奠定了基础。     

响应曲面法优化Lipozyme TL IM催化大豆油制备1,3-甘油二酯

为优化固定化脂肪酶Lipozyme TL IM催化大豆油制备高纯度1,3-甘油二酯,在单因素实验的基础上,选择催化温度、酶添加量、水添加量作为自变量,1,3-甘油二酯产率作为响应值,利用Central Composite Design响应曲面设计方法,研究各反应条件对1,3-甘油二酯产率的影响.再利用Design Expert模拟二次多项式回归方程的预测模型,确定最佳条件组合.结果表明:在酶解温度为55℃,加水量30％,加酶量45％,1,3-甘油二酯在产率为56.82％,1,2-甘油二酯产率为12.45％,甘油二酯合计69.86％.     

乳化剂对无溶剂体系合成1，3-甘油二酯的影响

在无溶剂体系中,通过脂肪酶Novozyme 435催化甘油解法合成功能性油脂1,3-甘油二酯。在底物摩尔比1∶1(甘油与天然谷物调和油比),脂肪酶添加量8%(占底物质量)条件下,研究酶催化反应过程中不同HLB值的蔗糖脂肪酸酯和反应温度对1,3-甘油二酯生成量的影响。研究结果表明,选择HLB值为11的亲水性蔗糖脂肪酸酯,在添加量0.5%,反应温度60℃下反应24 h,1,3-甘油二酯生成量最高,为42.61%。     

固定化脂肪酶催化酱油糟油脂合成甘油二酯的研究

本文以酱油糟为研究对象,利用实验室自主研发的低温连续相变萃取技术提取其中的油脂,并对提取出来的油脂主要理化指标进行分析,选用自制的具有Sn-1,3位专一性固定化脂肪酶HM IM对其油脂中丰富的游离脂肪酸(含量为61.50%)酯化合成甘油二酯。通过酶的添加量、甘油添加量、反应温度、搅拌速度、金属离子和不同辅助脱酸处理这几个因素进行优化,得到最佳的酯化合成甘油二酯条件为:自制固定化脂肪酶00S添加量1%、甘油添加量为理论所需量的1.2倍、反应温度65℃、真空度0.098 MPa+氮吹,2 mmol K+处理固定化酶、搅拌速度120 r/min下反应10 h,酱油糟油脂的游离脂肪酸酯化率达96.36%,甘油二酯含量达48.65%(43.65%的1,3-DG、5.00%的1,2-DG)、2.24%FFA、9.31%的MG,39.80%的TG,而且酯化反应不会改变油脂的脂肪酸组成。酶法酯化处理既降低了酱油糟油脂的酸值、又富含功能性甘油二酯,对其开发再利用提供新的途径。     

部分省份食用植物油中脂肪酸氯丙醇酯含量水平调查分析

研究调查了10个植物油品种的491批次食用植物油样本中3-氯-1,2-丙二醇脂肪酸酯(3-MCPD酯)、2-氯-1,3-丙二醇脂肪酸酯(2-MCPD酯)、1,3-二氯-2-丙醇脂肪酸酯(1,3-DCP酯)和2,3-二氯-1-丙醇脂肪酸酯(2,3-DCP酯)4种脂肪酸氯丙醇酯的含量水平,为监管工作提供基础数据。用间接测定法以七氟丁酰基咪唑(HFBI)为衍生剂,GC-MS测定,多同位素内标法定量。调查结果显示:所有样本均未检出1,3-DCP酯和2,3-DCP酯;3-MCPD酯检出率为83.3%,检出结果范围为0.104~8.580 mg/kg;2-MCPD酯检出率为66.0%,检出结果范围为0.106~4.320 mg/kg;3-MCPD酯和2-MCPD酯的同时检出率为65.2%;脂肪酸氯丙醇酯含量水平较高的油脂品种是棕榈油和油茶籽油,含量水平较低的油脂品种是橄榄油和大豆油;不同植物油品种脂肪酸氯丙醇酯含量水平差异较大。     

脱臭温度下油脂成分对3-氯丙醇酯和缩水甘油酯形成的影响及其形成机理的推测

摘要：以甘油一酯（MAG）、1,2-甘油二酯（1,2-DAG）、1,3-甘油二酯（1,3-DAG）、甘油三酯（TAG）和四丁基氯化铵（TBAC）纯品作为前体物质,正十六烷为基质模拟化学体系,于氮气保护下240 ℃加热2 h以模拟油脂脱臭过程,考察甘油酯类型和游离脂肪酸对3-氯丙醇酯（3-MCPDE）和缩水甘油酯（GEs）形成的影响。结果表明：3-MCPDE和GEs含量与MAG、DAG含量呈正相关,而与TAG含量关系不大,其中MAG的影响最大;在MAG体系中,3-MCPDE和GEs含量与游离脂肪酸含量呈正相关,在1,2-DAG和1,3-DAG体系中,游离脂肪酸含量对3-MCPDE和GEs含量的影响较小,而在TAG体系中,游离脂肪酸含量对3-MCPDE和GEs含量基本无影响;除TAG体系外,在添加脂肪酸的MAG和DAG体系中,3-MCPDE含量远高于未添加脂肪酸的体系;在游离脂肪酸含量相同时,不同甘油酯类型的体系中3-MCPDE和GEs含量大小分别为MAG>1,2-DAG>1,3-DAG>TAG和MAG>1,3-DAG>1,2-DAG>TAG。     

氯丙醇检测方法研究进展

氯丙醇类化合物是国际公认的食品污染物,包括3-氯-1,2-丙二醇(3-MCPD)、2-氯-1,3-丙二醇(2-MCPD)以及双氯取代的1,3-二氯-2-丙醇(1,3-DCP)和2,3-二氯-1-丙醇(2,3-DCP)。文章综述了国内外氯丙醇检测方法的研究进展,包括气相色谱(GC)法、气质联用(GC/MS)法、气相色谱双串联质谱(GC/MS/MS)联用法以及其它新的检测方法。     

橄榄油甘油二酯异构体的HPLC-ELSD测定方法建立及其应用

目的:建立一种用于测定橄榄油中甘油二酯异构体的方法,并应用于评价橄榄油的新鲜度。方法:采用正相高效液相色谱法,以异丙醇-正己烷为流动相进行梯度洗脱,应用蒸发光散射检测器进行检测。结果:1,3-甘油二酯(1,3-DAG)和1,2-甘油二酯(1,2-DAG)在20 min内实现分离,分别在11.16～111.62 μg/mL和9.50～95.00 μg/mL范围内线性关系良好,平均回收率分别为103.2%和101.4%,重复性及日间精密度RSD均低于4.0%;应用该方法测定了6批特级橄榄油的甘油二酯异构体含量,测定结果与橄榄油氧化情况相关性较好。结论:该方法能够准确、快速地测定橄榄油甘油二酯异构体的含量。     

The polymorphism of diglycerides

The polymorphism of 1,3-dipalmitin, 1,2-dipalmitoyl-sn-glycerol, 1.2-dipalmitoyl-rac-glycerol has been examined by X-ray diffraction, IR spectroscopy and Raman spectroscopy and differential scanning calorimetry. The respective spectra and thermographs are presented for these polymorphs. Optically active 1,2-diglycerides exhibit very different thermal behaviour from the optically inactive 1,2-diglycerides. A novel HPLC system was devised for the separation of the diglycerides.     

The characteristics of percolation olive oils produced with a new processing enzyme aid

Theee olive varieties ( Leccino , Coratina and Dritta ) were processed using the percolation–centrifugation system with a new olive enzyme processing aid, 'Cytolase 0'. The enzyme adjuvant led to oils characterized (with respect to reference oils) mostly by: (i) higher contents of total phenols, o -diphenols, tyrosol- and hydroxytyrosol-aglycones, tocopherols, trans -2-hexenal and other pleasant aromatic volatile compounds, and triterpene alcohols; (ii) higher values of oxidative stability and carotenoid colour index; (iii) higher ratios of 1,2-diglycerides/1,3-diglycerides, campesterol/stigmasterol, trans -2-hexenal/hexanal, and trans -2-hexenal/total aroma; (iv) lower values of turbidity; (v) similar values of acidity, peroxide index, carbonyl index, UV (ultraviolet) spectrophotometric indices and alcoholic index; (vi) similar contents of saturated fatty acids, unsaturated fatty acids, triglycerides, diglycerides, sterols, and green lipochromes; and (vii) similar sensory score. Stigmastadienes, trans -oleic, trans -linoleic, and trans -linolenic acid isomers were not detected. Hence, the enzyme adjuvant produced better qualitative characteristics and higher extraction outputs.     

延黄牛脂制备高纯度1,3-甘油二酯工艺

目的:提高延黄牛脂的附加值。方法:采用固定化脂肪酶Lipozyme RM IM为催化剂,牛油脂肪酸乙酯(乙醇解法自制)和甘油为原料制备1,3-甘油二酯,运用氢谱考察脂肪酶添加量、底物摩尔比、反应时间以及反应温度对粗反应混合物中1,3-甘油二酯含量的影响,并阐明醇解酯交换前后牛油与甘油二酯产物的脂肪酸组成变化。结果:1,3-甘油二酯合成最佳条件为脂肪酶Lipozyme RM IM质量分数为1%,底物摩尔比(n_{脂肪酸乙酯}∶n_甘油)2∶1,反应时间6 h,反应温度50℃。此条件下的1,3-甘油二酯生成率为72.5%,甘油酯得率为77%;纯化后纯度提高至90.79%。与原油(延黄牛脂)相比,甘油二酯产物中亚油酸和油酸含量分别升高了13.65%,6.47%,饱和度降低了7.17%。结论:延黄牛脂制备1,3-甘油二酯不仅能改变甘油酯结构,还可以改变牛脂的脂肪酸组成,降低其饱和度。     

Influence of diglycerides on crystallisation of palm oil

Palm oil diglycerides and pure synthetic diglycerides of 1,2‐(2,3‐) and 1,3‐dipalmitoylglycerol, dioleoylglycerol and palmitoyloleoyl glycerol were added to purified triglycerides of palm oil. The crystallisation kinetics of the samples were studied by spectrophotometry, NMR analysis of the solid fat content and by X‐ray diffraction of the crystals obtained at 20°C. Palm oil diglycerides inhibited the nucleation process and retarded the crystal growth rate of palm triglycerides. The effect was less pronounced at a higher degree of supercooling. The extent of inhibition depended on the nature of the diglycerides and the concentration. © 1999 Society of Chemical Industry     

A method based on 1H NMR spectral data useful to evaluate the hydrolysis level in complex lipid mixtures

A new approach is reported, based on Proton Nuclear Magnetic Resonance (¹H NMR) spectral data, to study qualitatively and quantitatively the hydrolysis level in complex lipid mixtures. It is based on a detailed study of the spectra of tri-, di-, mono-glycerides and fatty acids. It has been validated by using mixtures of standard compounds which simulate the complex mixtures formed in lipid digestion. To this aim the ¹H NMR spectra of pure compounds, as well as of mixtures containing different known proportions of triglycerides, 1,2-diglycerides, 1,3-diglycerides, 1-monoglycerides, 2-monoglycerides and fatty acids, were acquired. These mixtures cover a very broad range of concentrations and simulate edible oils and fats of vegetable and animal origins, including fish, with different levels of hydrolysis. Equations based on spectral data are proposed to determine the molar percentage of the different kinds of compounds that may be formed in lipid digestion, thus allowing quantification of the advance of lipolysis. To the best of our knowledge, this is the first time that a method based on ¹H NMR spectral data to quantify triglycerides, as well as partial glycerides and fatty acids in complex lipid mixtures coming from lipolysis is proposed. This methodology offers many advantages and is useful not only in the fields of food technology and nutrition, but also in others like enzymology, pharmacology, medicine and petrochemistry, among others.     

Solvent-free production of 1,3-diglyceride of CLA: Strategy consideration and protocol design

Enzymatic production of a homogeneous 1,3-diglyceride of polyunsaturated fatty acids (PUFAs) was carried out using Novozym 435 as biocatalyst and conjugated linoleic acid (CLA) as a model fatty acid. Three different operation modes, namely, magnetic stirring under vacuum, vacuum-driven N₂ bubbling and incubation with molecular sieves, were examined to find an efficient protocol for the enzymatic production. Studies on the effects of mass transfer showed that the occurrence of mass transfer limitation was strongly dependent on the operational modes. Vacuum-driven N₂ bubbling proved to be capable of eliminating mass transfer resistance, creating effective interaction for a multiple-phase reaction system and yielding an efficient water removal and a faster reaction rate. Hence, vacuum-driven N₂ stirring was considered as the best choice among the tested strategies for the production of pure 1,3-diglyceride of PUFAs with industrial interests, because it gave a higher yield of the desired product, higher productivity and lower impurity content due to its suppression of the acylmigration of 1,3-diglyceride to 1,2-diglyceride. The yield of 92–96% 1,3-dCLG could be obtained when 5 mmol of glycerol were incubated with 10–12 mmol CLA for about 3 h at 45–55 °C and a pressure less than 10 mbar, with enzyme loading of 40–70 g l⁻¹. Among the operational parameters, temperature and reaction time were found to have profound effects on the acylmigration and yield of 1,3-diglyceride. Moreover, the enzyme showed excellent operational stability in this protocol under the optimized conditions (little activity loss of enzyme was observed after 10 consecutive batch reactions), indicating the potential of this technology for industrial application.     

Polymorphism of rapeseed oil with a low content of erucic acid and possibilities to stabilise the β′-crystal form in fats

The polymorphism of hydrogenated rapeseed oil with a low content of erucic acid was investigated by X-ray diffraction. The triglyceride composition in the hydrogenated oil was determined by a combination of gas chromatography and high-performance liquid chromatography. A characteristic feature of this oil is its rapid transition to the β-form. Possibilities of influencing the β′→ β transition are discussed. The addition of diglycerides appears to provide a means for delaying the β′rarr; β transition. Thus, 1,2-diglycerides show a strong stabilising effect on the β′ triglyceride crystals form, which is considered to be of technical interest.     

Lipolytic activity of Williopsis californica and Saccharomyces cerevisiae in extra virgin olive oil

Inoculation trials performed with three strains of yeasts, isolated from extra virgin olive oil, Williopsis californica 1,639, Saccharomyces cerevisiae 1,525 and Candida boidinii 1,638, demonstrated that some yeast can lower the quality of the oil during storage. Laboratory tests highlighted a substantial increase in the total diglycerides and free fatty acids in the samples of oil inoculated with the lipase-producing strains of yeasts, W. californica 1,639 and S. cerevisiae 1,525, while in the samples of oil inoculated with the lipase-negative strain C. boidinii 1,638 no differences were found in respect to the uninoculated control. The acidity of the extra virgin olive oil, inoculated with the lipase-producing strains W. californica 1,639 and S. cerevisiae 1,525, during 2 weeks of incubation at 30 degrees C increased respectively from 0.62% to 1.50 and 1.62%, exceeding the limit of 0.8% established by current regulations for this commercial category of olive oil, while in the oil inoculated with the lipase-negative strain and in the uninoculated control, the acidity remained constant throughout. Furthermore, the two strains of lipase-producing yeasts also increased the concentration of the 1.3-diglyceride isomer in the oil lowering the values of the total 1.2-diglycerides/total 1.3-diglycerides ratio considered to be an important index of quality for an extra virgin olive oil. The lipolytic activity of lipase-producing strains W. californica 1,639 and S. cerevisiae 1,525 showed an optimum pH of 6 and 7.5 and an optimum temperature of 20 degrees C and 30 degrees C respectively. Nevertheless, the lipolytic activity was negatively influenced by glucose and polyphenols when the concentration was higher than 0.25% and 0.4% (wt/vol) respectively.     

Characteristics of Mackerel Mince Lipid Hydrolysis

The lipid hydrolysis products in minced mackerel stored under vacuum at 2–3°C for 15 days were free fatty acids (FFA) and 1,2-diglycerides (DG); no 1,3-DG br monoglycerides were obsrved, Lysophosphati-dylcholine fLPC and lvsoohosohatidvlethanolamine (LPE) increased and then ddcreased with time: Lysiphosphatidylserie was not observed. More FFA, 1,2-DG, LPC, and LPE were detected in dark muscle than in light muscle. The initial fatty acid composition, by GC, showed no significant differences (α-0.05) among whole, light and dark muscles. When the lipid extracts were separated into three fractions (1: mostly triglycerides; 2: mostly FFA, DG, and cholesterol; and 3: mostly phospholipid), the degree of unsaturation of the lipids in fraction 2 was between those in fraction 1 and 3 and increased with time. These findings support the possibility that fish lipid hydrolysis may affect fish wholesomeness.     

食品中氯丙醇脂肪酸酯风险及应对措施概述

氯丙醇酯作为食品中的一类新型污染物,得到世界各国和地区的广泛关注和研究。氯丙醇酯类物质种类繁多,其反应机制、毒理学和代谢转化研究尚不成系统,目前研究表明氯丙醇酯类物质具有一定的肾脏、生殖毒性及潜在的致癌性。国内外对氯丙醇酯的检测方法、污染水平和暴露评估等研究主要集中在3-氯-1,2-丙二醇酯和2-氯-1,3-丙二醇酯,且主要集中在婴儿配方奶粉、油炸和烘焙食品等脂肪含量较高的食品上。通过研究发现,普通消费者从日常饮食中摄入氯丙醇酯类物质一般是无风险的,但对于一些年轻群体和仅食用配方奶粉的婴幼儿来说风险较高。国内外已相继开展了对氯丙醇酯的研究和监控,以减少食品中氯丙醇酯类物质污染水平。目前只有欧盟制定了3-氯-1,2-丙二醇酯在部分食用油和婴幼儿配方食品中的限量要求,仅国际食品法典委员会制定了植物油中3-氯-1,2-丙二醇酯的控制规范,来降低氯丙醇酯的风险。鉴于此,本文主要对氯丙醇酯形成机制、代谢、毒性、国内外食品中氯丙醇酯的污染水平、风险评估研究进展及其管理控制措施等方面进行了简要概述,以期为食品安全监管提供参考。