草原苏尼特羊脑中磷脂型DHA萃取及其结构特性研究

旨在为绿色天然草原羊脑中磷脂型二十二碳六烯酸(DHA-PL)的开发利用提供科学数据,以苏尼特冻干羊脑为原料,采用溶剂萃取其中的粗脂肪,用冰丙酮沉淀得到磷脂,再采用硅胶柱层析法分离其中的磷脂酰胆碱(PC)和磷脂酰丝氨酸(PS)。测定了粗脂肪中胆固醇含量,以及磷脂中PC和PS的含量,分析了粗脂肪、磷脂、PC和PS中DHA含量,以及PC和PS中sn-2位的DHA含量。结果表明:采用体积比2∶1乙醇-正己烷萃取冻干苏尼特羊脑中粗脂肪,粗脂肪得率较高,为45%;羊脑粗脂肪中胆固醇含量为29.42mg/g,粗脂肪的脂肪酸组成中DHA含量为8.89%;磷脂的脂肪酸组成中DHA含量为11.41%;磷脂中PC和PS含量分别为27.67μg/g和10.69μg/g,PC和PS的sn-2位脂肪酸组成中DHA含量分别为8.06%和2.59%。综上,可以从苏尼特羊脑中制备易于吸收的DHA-PL。

Synthesis,Crystal Structure and Antitumor Activity of a New 3H-Phenanthro-[2,1-d]imidazole Derivative of Dehydroabietic Acid

The title compound （CI9H24BrN302, 3） was synthesized from dehydroabietic acid and its crystal structure was determined by single-crystal X-ray diffraction. The compound is of monoclinic system, slSace group C2 with a = 24.830（5）, b = 7.1410（14）, c = 13.981（3） A, p = 107.68（3）°, Z = 4, V= 2361.9（8） A3, Mr = 482.42, Dc = 1.357 Mg/m3, S = 1.003,μ = 1.772 mm1, F（000） = 1008, the final R = 0.0660 and wR = 0.1402 for 2244 observed reflections （I 〉 2o（/））. The crystal structure is stabilized by four hydrogen bonds （O（W）-H（WA）...O（3）, O（W）-H（WB）...O（1）, N（2）-H（2A）...O（W） and N（3）-H（3A）...N（1）） formed among the title compound, the crystal water and acetone molecules. The preliminary antitumor assay showed the title compound exhibited considerable inhibitory activity against HepG2 and SMMC-7721 cell lines with the ICs0 values of 17.1 and 10.2 μM, respectively.

Syntheses and Crystal Structures of 12-Oxime Dehydroabietic Acid Derivatives

Three novel dehydroabietate oxime derivatives were synthesized from dehydroabietic acid and their crystal structures were determined by X-ray crystallographic techniques. All of these 12-oxime dehydroabietic acid derivatives crystallize in orthorhombic system, space group P212121. In the structures, rings A and B exhibit chair and half-chair configurations, respectively and form trans ring junction with two methyl groups（C（19） and C（20）） in the axis positions. The oxime groups have E conformations. Comparison of conformations reveals subtle differences principally in ring B due to the modification in C（12）. The E-acetaldehyde oxime derivative 2c showed distinct BKα gate-opening activity with an ionic current increase of 164% at 30 μM versus the control current.

Synthesis,Crystal Structure and Cytotoxic Activity of a New N-Vinyl-1H-dibenzo[a,c]carbazole Derivative of the Dehydroabietic Acid

The title compound （C29H33NO2, 3） was synthesized and structurally characterized by elemental analysis, IR, MS, 1H- and 13C-NMR and single-crystal X-ray diffraction. The crystal is of monoclinic system, space group P21 with a = 14.428（3）, b = 7.3440（15）, c = 22.768（5） A, β = 95.17（3）°, V = 2402.7（8） A3, Z = 4, Mr = 427.56, Dc = 1.182 g/cm3, F （000） = 920, 2（MoKa） = 0.71073 A,μ = 0.073 mm-1, the final R= 0.0670 and wR= 0.1002 for 2437 reflections with I〉 2σ（I） Two crystallographically independent molecules with different conformations co-exist in the structure. The crystal structure is stabilized by intermolecular C-H…π interactions which make the molecules stack along the b axis. In addition, the preliminary cytotoxic assay showed that the title compound exhibited moderate inhibitory activity against KB and SW1116 cells.

Dehydroabietic acid chemosensitizes drug-resistant acute lymphoblastic leukemia cells by downregulating survivin expression

Objective:To explore the mechanism of drug resistance in acute lymphoblastic leukemia and the anti-tumor effect of combination therapy of dehydroabietic acid and vincristine against acute lymphoblastic leukemia cells.Methods:Acute lymphoblastic leukemia cells REH and CCRFCEM were employed to detect the anti-tumor effect of vincristine and doxorubicin on proliferation and apoptosis using EdU assay,human active caspase-3 Quantikine ELISA kit,and flow cytometry.Vincristine-resistant REH cells(REH-R),survivin knockdown and overexpressing REH cells were established to verify the role of survivin in drug resistance.Additionally,in vitro and in vivo assays were performed to determine the effect of dehydroabietic acid on the cytotoxicity of vincristine.Results:Vincristine and doxorubicin markedly suppressed proliferation and induced apoptosis of REH and CCRF-CEM cells.Survivin expression was upregulated in REH-R cells compared with REH cells.Knockdown of survivin expression obviously restored the sensitivity of REH-R cells to vincristine.Akt phosphorylation was also increased in REH-R cells compared to REH cells.In addition,LY294002,a PI3k/Akt pathway blocker,inhibited survivin expression and enhanced cytotoxicity of vincristine to REH-R cells.Dehydroabietic acid effectively reduced survivin expression in REH-R cells,thereby enhancing the therapeutic effect of vincristine on drug-resistant cells.Survivin overexpression markedly reduced the effect of dehydroabietic acid on enhancing the anti-proliferation and inducing apoptosis effect of vincristine.Moreover,the combination of dehydroabietic acid with vincristine significantly extended the survival rate in a mouse xenograft model of acute lymphoblastic leukemia,compared with vincristine treatment alone.Conclusions:Dehydroabietic acid may be used as a potential candidate for the treatment of acute lymphoblastic leukemia in combination with vincristine.

藻油DHA微胶囊制备及稳态化递送技术研究进展

藻油DHA为多不饱和脂肪酸,具有促进大脑生长发育,预防心血管疾病,参与免疫调节等多种生理功能,然而其极不稳定,易受到光、热、水及CO_(2)等因素的影响,导致其氧化、变质。目前,国内外对藻油DHA稳态化递送技术的研究颇多,微胶囊技术是其中一种较为成熟的方法,且应用最为广泛,能够有效增强藻油DHA的递送能力、贮藏稳定性及生物利用度。本文概述藻油DHA的来源、理化性质、生物活性及其递送体系等,重点探讨藻油DHA微胶囊稳态化靶向递送技术,包括藻油DHA微胶囊的制备方法、递送体系、递送机制、影响因素等,旨在为拓宽藻油DHA在食品领域的应用范围,提供一定的理论依据。

Two Dehydroabietic Acid-based Arylamines: Synthesis, Crystal Structure and Fluorescent Properties

Two dehydroabietic acid-based arylamines have been synthesized and characterized by FTIR, 1H NMR, 13C NMR, MS spectra and elemental analysis. Their spatial structures were determined by X-ray diffraction analysis. UV-Vis absorption and fluorescence spectral characteristics of these compounds in methanol were investigated. Their fluorescence emission spectra in different polarity solvents were further evaluated. Fluorescent properties and structural relationship of the compounds showed that fluorescence intensity and quantum yield inversely increase with the non-coplanar degree. In addition, the solvent polarity has different effects on the fluorescence emission spectra of two compounds.

微藻破囊壶菌产功能性脂肪酸DHA研究进展

二十二碳六烯酸(DHA)作为人体必需的多不饱和脂肪酸,在维护心血管健康、抗癌、支持视觉和脑功能等方面至关重要。传统的深海鱼油提取DHA方法存在鱼腥味重、工艺繁琐等问题,迫使研究者寻求更为高效、环保的替代方案。破囊壶菌(Thraustochytrids)凭借其生长迅速、低重金属污染以及高DHA含量的特性,成为工业化生产DHA的潜力微生物之一。当前在破囊壶菌发酵生产DHA的过程中,依然需要解决一系列关键问题,包括提高发酵产量、降低成本等。本文旨在全面阐述破囊壶菌发酵生产DHA的研究现状,包括菌株筛选与改良、DHA生物合成途径、遗传转化及代谢工程、发酵控制策略等方面。首先,总结归纳了对野生型菌株的自然筛选和诱变改良等方法,不断提高破囊壶菌中DHA产油量。其次,详细介绍了破囊壶菌DHA合成途径的研究进展,着重分析了生物合成途径中关键辅助因子在DHA生产中的作用。此外,概述了外源DNA传递到破囊壶菌细胞的遗传转化技术的应用现状,为提高其遗传转化效率和稳定性提供重要参考。在DHA代谢调控方面,探讨了氮限制对DHA合成的促进作用以及温度和氧气供应对生产效率的影响。最后,对利用破囊壶菌生产DHA存在的主要瓶颈问题和未来发展趋势进行了总结,以推动其在医药、保健品和食品等领域的广泛应用,实现工业规模下的高效生产。

Synthesis and Crystal Structure of a Novel Lactone Derived from Dehydroabietic Acid

A novel lactone derived from dehydroabietic acid,C20H21BrO4,has been charac-terized by IR,1H NMR,13C NMR and single-crystal X-ray diffraction method.It crystallizes in orthorhombic,space group P212121 with a = 6.4195（15）,b = 11.535（3）,c = 24.654（6） ,V = 1825.5（7） 3,Z = 4,Mr = 405.28,Dc = 1.475 g/cm3,λ = 0.71075 ,μ（MoKα） = 2.27 mm-1,F（000） = 832,the final R = 0.024 and wR = 0.045 for 2152 observed reflections with I 2σ（I）.The molecules of lactone are mainly linked through intermolecular hydrogen bonds.

DHA的微囊化制备、应用与吸收利用研究

二十二碳六烯酸(docosahexaenoic acid,DHA)作为一种多不饱和脂肪酸,由于其低溶解度、低稳定性,导致其在食品、保健品的应用和人体的吸收利用受到限制。本研究采用喷雾干燥技术制备DHA微囊粉,评价其在软糖中的应用,对比在加速稳定性实验中挥发性物质的含量变化。利用Caco-2细胞单层模型,考察各产品释放终产物的表观渗透系数(Apparent permeability coefficient,Papp),对比不同剂型之间的体外生物利用度。DHA微囊粉的包埋率为99.98%,DHA含量为11.2%。与DHA藻油相比,具有较好的腥味掩盖能力,能够较好应用到明胶软糖中,Papp系数提高2.23倍。通过微囊化技术制备的DHA微囊粉具有包埋率高、粒径小、水溶性高等特点,能够显著降低DHA在应用过程中的腥味物质以及提高其体外生物利用度。

磷脂型DHA的消化、吸收与转运

二十二碳六烯酸(DHA)参与大脑生长发育,影响大脑中神经递质、突触可塑性以及信号转导等生理功能。目前,市售DHA膳食补充剂产品较多,其脂质结构主要分为甘油三酯型DHA(TAG-DHA)、乙酯型DHA(EE-DHA)和磷脂型DHA(PL-DHA)。DHA不同酯化结构影响其吸收、转运和生物利用度。目前PL-DHA和TAG-DHA生物利用度高仍存在争议,而大部分的研究证明PL-DHA是最佳的DHA吸收形式,尤其是能被脑组织更好地吸收。本文比较分析磷脂型DHA与其它形式DHA的吸收、转运等,总结不同模型中磷脂型或甘油三酯型DHA在人体或动物体内生物利用度。目前大多是动物实验研究,还需更深入的人体试验,来探究PL-DHA在人体的吸收过程及其机理,评价其生物利用度以及对神经发育的长期作用。PL-DHA有望成为一种更高效的DHA食品营养强化剂,添加到婴儿配方奶粉中或制成胶囊服用。

Preparation of DHA-and EPA-enriched Glycerides by Enzymatic Interesterification Using Tuna Oil and Capric Acid

This study focused on the preparation of docosahexaenoic acid(DHA) and eicosapentaenoic acid(EPA)enriched-triacylglycerols by enzymatic interesterification using tuna oil and capric acid. The content of DHA+EPA is 26.86% in the tuna oil used in this study. A response surface methodology(RSM) was used to optimize the reaction parameters(reaction temperature, substrate molar ratio, enzyme amount and reaction time), and the optimized conditions were determined to be: reaction temperature 58℃, substrate molar ratio(capric acid : tuna oil) 4:1, enzyme amount 4%, and reaction time 7.5 h. Under the optimized conditions, the content of DHA+EPA in the glycerides was 40.03%, which is 13.17% higher than that in raw tuna oil. In addition,the MLM-type structured lipids containing medium chain fatty acids(capric acid) at positions sn-1,3 and a long chain fatty acid(DHA/EPA) at the position sn-2 may have many health benefits for humans.

DHA营养强化鸡蛋研究进展

DHA能有效促进大脑发育、改善视力、预防心血管系统疾病等,但其在人体内的生成量不能满足需求,需要靠食物获得。开发DHA营养强化鸡蛋,可以为人们获取足量、廉价、优质的DHA提供有效途径。为了加强对DHA营养强化鸡蛋的研发,推动其进一步发展,从DHA的生理功能视角出发,分析了DHA营养强化鸡蛋开发优势、饲料脂肪酸对鸡蛋中DHA的调控机制,阐述了DHA营养强化鸡蛋的脂质组成、饲料中DHA的来源及鸡蛋中DHA的稳定性。富含DHA的日粮可生产富集DHA的鸡蛋,且蛋黄中的DHA具有较高的稳定性,同时DHA营养强化鸡蛋食用方便、便于保存,且价格相对低廉,因此DHA营养强化鸡蛋具有广阔的发展前景。

菜籽油-DHA藻油调和油在不同鸡肉烹饪方式下的品质变化

为了对DHA藻油调和油的开发和合理烹饪提供相关指导,将DHA藻油分别按菜籽油质量的2%、3%、5%、10%、15%与菜籽油复配制成菜籽油-DHA藻油调和油,并以蒸、炒、烧三种不同方式烹饪鸡肉,测定烹饪前后油样的脂肪酸组成、酸值和过氧化值的变化,分析探讨DHA藻油比例及烹饪方式对调和油脂肪酸组成及理化指标的影响。结果表明:烹饪后,调和油的单不饱和脂肪酸含量减少,饱和脂肪酸和多不饱和脂肪酸含量增加;三种主要脂肪酸油酸、亚油酸、亚麻酸含量随DHA藻油比例增加呈不规律波动变化;除DHA藻油比例为2%的调和油外,其余调和油的酸值均在烹饪后增加,其中,DHA比例为3%、5%和10%的调和油炒鸡肉后酸值增加最大,蒸和烧这两种烹饪方式对酸值的影响相对较小;纯菜籽油、DHA藻油比例为15%的调和油的过氧化值增加量明显大于其他的调和油,总体上炒较另外两种烹饪方式更易导致过氧化值增加。综上,建议蒸鸡肉或烧鸡肉宜采用DHA藻油比例为2%、3%、5%的调和油,炒鸡肉则不宜采用DHA藻油调和油。

尿素包合法联用分子蒸馏法富集鱼油中EPA和DHA的研究进展

鱼油中的EPA和DHA是一类具有很高营养价值的功能因子,对心脑血管健康、大脑发育、改善视力、抗炎等都有积极的作用,因而获取高纯度的EPA和DHA逐渐成为众多研究者关注的热点问题之一。采用尿素包合法富集鱼油中的EPA和DHA,不仅原料成本低廉,而且操作简单,但是富集得到的鱼油质量不高;而分子蒸馏法可以较好地保护EPA和DHA的活性,提高鱼油的纯度。两种方法联用具有工艺成熟、原料成本低、操作简单等优势,是工业化生产高质量EPA和DHA较好的选择之一。因此,在总结富集纯化鱼油中EPA和DHA方法的同时,阐述了尿素包合法、分子蒸馏法及两种方法联用富集鱼油中EPA和DHA的研究进展,希望可以为未来工业化获取高质量的EPA和DHA提供一定的参考价值。

Matrix Extension with Fitness for Purpose and Stability Assessment of DHA and Additional Fatty Acids in Individual Whole Chicken Eggs

The consumption of long chain polyunsaturated fatty acids (LC-PUFA) is associated with several human health benefits. Most notable of these LC-PUFA is docosahexaenoic acid (DHA C22:6) whose inclusion is considered essential for optimum human health. Biofortification of common foods such as eggs with DHA has emerged as a specific approach to increase the intake of DHA in human populations. This can be achieved by supplementing poultry rations with feeds like microalgae or fish oil that are rich in DHA, which results in an increased uptake in the egg. Gas chromatography with flame ionization detection (GC-FID) is the method of choice when analyzing food such as eggs for DHA and other fatty acids. For regulatory studies it is desirable to demonstrate that the method is specifically suitable for the analysis of DHA and fatty acids in eggs. The purpose of this paper is to further extend the scope of the AOAC 996.06 methodology examined in the paper by Dillon et al., and to demonstrate the fitness for purpose of the method by examining specific validation parameters. It is a further objective to investigate the stability of DHA and other fatty acids of short and long timepoints. A validation of the method for the determination of DHA and three other fatty acids in eggs is thus presented.

Analytical Method Assessment for the Determination of DHA and Fatty Acids Present in Unextracted <i>Aurantiochytrium limacinum</i>Biomass

Research into long-chain polyunsaturated fatty acids (LC-PUFA), such as docosahexaenoic acid (DHA C22:6 n-3), has shown that their inclusion in the human diet is linked with many health benefits. This has led to an increased interest in the enrichment of certain foodstuffs with DHA by supplementing animal fed with DHA-rich ingredients which can lead to an increased uptake in the meat, milk and eggs animal by-products. The microalgae Aurantiochytrium limacinum has been found to be especially useful in this pursuit. It is subsequently desirable to availably have a simple and robust method for the routine analysis of DHA and other fatty acids in the algal biomass. The AOAC method 996.06 is often followed for the analysis of fatty acids in foods and demonstrating that its fitness for purpose in the analysis of DHA and additional fatty acids in Aurantiochytrium limacinum is therefore the objective of this paper. A validation of the method for the determination of DHA and three other fatty acids in Aurantiochytrium limacinum is presented. The method was found to be linear over the following ranges for each fatty acid methyl ester (FAME) analyte;50 to 15,000 μg/ml (C14:0), 300 to 95,000 (C16:0), 25 to 15,000 (C18:0) and 300 to 59,375 (C22:6). The accuracy, precision and LOD and LOQ of the method were confirmed and its robustness tested. The application of the method to assess the stability of Aurantiochytrium limacinum containing two alternative antioxidants was further examined. The investigation showed that DHA was stable over six months with the inclusion of either Duralox? or ethoxyquin as an antioxidant and ethoxyquin could additionally stabilize DHA in Aurantiochytrium limacinum up to 24 months.

利用Crypthecodinium cohnii高密度发酵生产DHA的流加策略研究

利用流加策略实现了隐甲藻(Crypthecodinium cohnii)生产DHA的高密度发酵。根据隐甲藻间歇式发酵的特性,探讨最佳补料时间及流加液中最佳碳氮比,结果表明,在葡萄糖浓度降至4g/L左右时流加碳氮比为30∶1的营养液对隐甲藻的生长和脂肪酸积累最有利。根据以上结果,初始葡萄糖浓度25g/L,采用5d连续发酵多次流加策略,细胞干重达6.37%,DHA产量为4.08g/L。

脂肪酶富集DHA和EPA的研究进展

综述了脂肪酶在富集DHA和EPA方面的应用 ,总结了近十年来脂肪酶富集DHA和EPA的研究状况 。

尿素包合法富集鱼油中的EPA和DHA的研究

用尿素包合法（脲包法）对大连鱼油中的ＥＰＡ和ＤＨＡ进行富集，得到较佳操作条件为温度０℃，脲酯比１３，鱼油甲酯为脲包客体，以甲醇作溶剂。