基于荧光及紫外光谱法对虾青素与乳清蛋白相互作用的研究

本研究采用分子自组装技术制备虾青素/乳清蛋白纳米复合物,并探究虾青素（Astaxanthin,AST）与乳清蛋白（Whey Protein）之间的分子相互作用机制。选用乳清蛋白（α-乳白蛋白、β-乳球蛋白、浓缩乳清蛋白、牛血清蛋白）与虾青素自组装,通过控制水相蛋白质溶液与有机相虾青素溶液的比例,分别可控形成H聚集体或J聚集体虾青素/乳清蛋白纳米复合物（H/J Aggregates Astaxanthin/Whey Protein Nanocomplexes）。通过动态光散射仪（Dynamic Light Scattering,DLS）测定虾青素/乳清蛋白纳米复合物的粒径均在150~430 nm之间,多分散性指数（Polydispersity Index,PDI）表明分散性良好,电位在?12~?1 mV之间;采用透射电子显微镜（Transmission Electron Microscope,TEM）观察成功制备出的H聚集体或J聚集体虾青素/乳清蛋白纳米复合物呈边缘清晰光滑的近球形结构;紫外可见光谱表明4种蛋白质构建的H聚集体或J聚集体虾青素/乳清蛋白纳米复合物中虾青素H聚集体最大吸收波长λ_max由虾青素单体的λ_max480 nm蓝移至388 nm,虾青素J聚集体最大吸收波长λ_max光谱红移,并显示出519和556 nm左右的并肩峰;通过荧光光谱分析表明由于虾青素聚集体特定的结构使形成的虾青素/乳清蛋白纳米复合物的荧光强度都明显增强,乳清蛋白中疏水性氨基酸及疏水性区域暴露。本研究从虾青素及其聚集体的水分散性和乳清蛋白载体特性两方面探究,为其后续在食品药品领域的开发及应用提供理论依据。     

不同类胡萝卜素清除自由基能力的比较

比较不同结构类胡萝卜素清除自由基能力。分别以虾青素、β-胡萝卜素、番茄红素和叶黄素为对象,采用DPPH法、水杨酸法和邻苯三酚自氧法分别表征清除DPPH·自由基、羟基自由基和超氧阴离子自由基能力。结果表明类胡萝卜素具有较强的清除自由基能力,且清除能力均遵循剂量-效应关系;对DPPH·自由基、羟基自由基和超氧阴离子的清除能力大小关系为虾青素叶黄素β-胡萝卜素番茄红素。类胡萝卜素清除自由基能力优良,不同结构类胡萝卜素清除自由基能力有差异,此类功能性产品具有广阔的市场前景。     

类胡萝卜素之间的转化

类胡萝卜素是广泛存在与自然界中的一种天然色素,不同种类的类胡萝卜素具有不同的生理功能,本文描述了多种类胡萝卜素之间的转化,包括叶黄素酯向玉米黄素的转化,叶黄素向4-酮基叶黄素的转化,辣椒红色素向4-酮基辣椒红色素的转化,玉米黄素向虾青素的转化,β-胡萝卜素向斑蝥黄质的转化,以及叶黄素向隐黄质的转化。     

虾青素聚集体的研究进展

虾青素是一种天然的萜烯类不饱和化合物,其分子结构中含有2个β-紫罗酮环和11个共轭双键。虾青素是自然界中唯一能够通过人体血脑屏障的类胡萝卜素,具有许多对人类健康有益的生理活性。近年来,虾青素因其分子结构所具有极强的抗氧化性在食品、医药、生物、化妆品等领域备受关注。虾青素作为脂溶性小分子在一定环境中会自聚集,从而呈现出不同的存在形式,如黄色的H聚集体、橘色的M单体、粉紫色的J聚集体,不同聚集体的颜色、结构、光学特性、生理活性等都会有巨大差异。本文主要对近十年来虾青素聚集体的形成机理、影响因素、检测方法(紫外可见光光谱法、透射电镜观察法、圆二色光法、拉曼光谱法、荧光光谱法、分子模拟法以及其他研究方法)及应用前景进行总结,以期为今后虾青素聚集体在生理活性和应用方面的研究奠定基础。     

高效液相色谱法测定鸡蛋、鸡肉中五种类胡萝卜素物质

本文研究了鸡蛋、鸡肉中虾青素、叶黄素、斑蝥黄、玉米黄素、β-胡萝卜素等5种类胡萝卜素同时检测的HPLC方法。结果表明,5种类胡萝卜素在0.5~100 μg/mL线性范围内有较好的线性关系,决定系数R2均大于0.996,方法的检出限均在0.10~0.20 mg/kg之间,定量限在0.33~0.66 mg/kg之间。在1、10、20 mg/kg的加标水平下,鸡蛋和鸡肉样品的加标回收率分别在71.6%~112.9%和88.6%~119.7%之间,RSD均≤7.2%。该方法快速简单、准确度高、重复性好,适用于鸡蛋、鸡肉等复杂基质样品中5种类胡萝卜素类物质检测的要求,对于鸡蛋、鸡肉的市场监管具有重要意义。     

强力抗氧化剂——虾青素

虾青素(Astaxanthin)是一种类胡萝卜素,属于酮式类胡萝卜素,全称为3,3-二羟基-4,4'-二酮基-β,β'-胡萝卜素,分子式为C40H5204,分子量为596.86.     

类胡萝卜素清除自由基活性构效关系

为探讨类胡萝卜素结构对其自由基清除能力的影响,采用四种自由基清除方法分别测定β-胡萝卜素、叶黄素、虾青素、番茄红素的自由基清除能力,对十三种类胡萝卜素进行了量子化学计算,通过AM1法对其结构进行初步优化后,再用密度泛函方法(Density functional theory,DFT)在B3LYP/6-311(d,p)水平上进一步优化计算。结果表明,不同种类类胡萝卜素对不同的自由基清除能力不同,虾青素最强,其次是番茄红素、叶黄素,β-胡萝卜素最弱;根据前线轨道能级差、NPA净电荷分布发现,类胡萝卜素多个共轭双键的特殊结构是其具有自由基清除能力的重要原因,其端环上的基团能够促进分子的活性,酮基是对其活性影响最大的基团,其次是羟基与环氧基,这与实验结果相吻合,表明前线轨道能级差、NPA电荷分布是表征类胡萝卜素自由基清除能力的重要参数。     

天然类胡萝卜素功能性产品的开发和应用

天然类胡萝卜素是一大类重要的食品着色添加剂、营养强化剂和抗氧化剂,本文概括了叶黄素、玉米黄质、虾青素、番茄红素和β-胡萝卜素的理化性质和卓越的生理功能,介绍了以上各类色素的加工工艺,展示了多种不同的主要剂型与在产品中的应用,说明了类胡萝卜素潜在的巨大的市场价值和提升空间。     

烹调方法对3种叶菜中类黄酮和类胡萝卜素的影响

目的:研究不同烹调方法对3种十字花科叶菜中总黄酮和类胡萝卜素的影响。方法:对娃娃菜、芥蓝、芥菜进行不同时间的焯煮、微波、蒸制,检测其中的总黄酮和3种类胡萝卜素含量。结果:总黄酮含量在焯煮后剧烈下降至53.1%,微波处理后升高。β-胡萝卜素蒸后损失可忽略,蒸5 min后叶黄素保存率在84%~97%之间;煮5 min后β-胡萝卜素保存率为78%~100%,叶黄素为84%~87%,而微波处理5 min后β-胡萝卜素保存率为60%~64%,叶黄素为77%~89%。结论:蒸制处理的保存效果最好,而微波处理后虽类胡萝卜素损失较大,但对保存类黄酮成分有益。     

虾青素抗氧化能力研究进展

虾青素是一种氧化型类胡萝卜素,主要来自微生物和海洋生物,因其特殊的化学结构而具有较强的抗氧化活性,从而被广泛应用于保健食品、医疗卫生等领域。本文从虾青素清除自由基能力、限制氧化剂渗透进细胞内、增加抗氧化酶活性、降低DNA的氧化损伤、抑制脂质过氧化5个方面阐述了虾青素的抗氧化机制,从体外清除自由基活性、油脂体系中抑制脂质氧化降解以及虾青素抑制促氧化酶、增强抗氧化酶活性3个方面阐述了虾青素的抗氧化能力,且综述了虾青素与其他天然抗氧化剂如番茄红素、叶黄素、β-胡萝卜素、维生素C和维生素E的抗氧化能力对比进展,为虾青素抗氧化活性的进一步研究提供参考。     

Major carotenoid composition of Brazilian Valencia orange juice: Identification and quantification by HPLC

The carotenoid composition of Brazilian Valencia orange juice was determined by open column chromatography (OCC) and high-performance liquid chromatography. Carotenoid pigments were extracted using acetone and saponified using 10% methanolic potassium hydroxide. Sixteen pigments were isolated by OCC and identified as α-carotene, ζ-carotene, β-carotene, α-cryptoxanthin, β-cryptoxanthin, lutein-5,6-epoxide, violaxanthin, lutein, antheraxanthin, zeaxanthin, luteoxanthin A, luteoxanthin B, mutatoxanthin A, mutatoxanthin B, auroxanthin B and trollichrome B. Thirteen carotenoid pigments were separated using a ternary gradient (acetonitrile–methanol–ethyl acetate) elution on a C₁₈ reversed-phase column. Among these, violaxanthin, lutein, zeaxanthin, β-cryptoxanthin, ζ-carotene, α-carotene, and β-carotene were quantified. The total carotenoid content was 12 ± 6.7 mg/l, and the major carotenoids were lutein (23%), β-cryptoxanthin (21%), and zeaxanthin (20%).     

Study of major aromatic compounds in port wines from carotenoid degradation

The carotenoids degradation and the formation of volatiles were examined by simulating Port wine aging. A two year old red Port wine was saturated with oxygen, supplemented with lutein and β-carotene and kept at 60 °C during 87 h. A similar study was performed in a model wine solution. Results showed that the percentage decrease in lutein levels was, respectively, 79% and 95%, in the wine model solution and in the Port wine, and 55% and 10% for β-carotene, indicating that lutein was more sensitive to degradation than β-carotene. Two other unknown degradation carotenoid compounds were identified by HPLC/DAD (reverse phase λ_max: 422; 445; 475 and 422; 445; 472) in the lutein supplemented wine. Levels of β-ionone and β-cyclocitral increased (2.5 times) in both, wine and wine model solution, supplemented with β-carotene. Along with these compounds, the same behaviour was observed in β-damascenone in the supplemented lutein wine and wine model solution. New insights were provided into the understanding of aroma modifications occurring during Port wine aging. The relationship between carotenoid molecules (β-carotene and lutein) and some volatiles has also been provided.     

Technical note: Simultaneous carotenoid and vitamin analysis of milk from total mixed ration-fed cows optimized for xanthophyll detection

Concentrations of retinol, α-tocopherol, and major carotenoids in dairy products are often determined simultaneously by liquid chromatography. These compounds have different polarity and solubility; thus, extracting them simultaneously can be difficult and inefficient. In milks with low carotenoid concentrations, the xanthophylls lutein and zeaxanthin may not be completely resolved using common extraction techniques. A simplified method was developed to optimize extraction efficiency and the limit of detection and limit of quantification (LoQ) of lutein and zeaxanthin in bovine milk without decreasing sensitivity to other vitamins or carotenoids. The developed method evaluates lutein, zeaxanthin, β-carotene, retinol, and α-tocopherol simultaneously by ultra-high performance liquid chromatography–photodiode array detection. Common saponification temperatures (40–60°C) and concentrations of KOH in water (10–50% KOH wt/vol) were evaluated. Multiple solvents were evaluated for optimal xanthophyll extraction (diethyl ether, dichloromethane, hexane, and tetrahydrofuran) following saponification. The limit of detection and LoQ were defined as 3:1 and 10:1 signal-to-noise ratio, respectively. All experiments were performed in triplicate. The optimal saponification procedure was a concentration of 25% KOH at either 40 or 50°C. Saponified extracts solubilized in solutions containing diethyl ether had greater concentrations of lutein- than hexane- or tetrahydrofuran-based solutions, with peak areas above LoQ values. The solution containing diethyl ether solubilized similar concentrations of retinol, α-tocopherol, and β-carotene when compared with other solutions. The proposed optimized method allows for the simultaneous determination of carotenoids from milk with increased lutein and zeaxanthin sensitivity without sacrificing recovery of retinol, α-tocopherol, and β-carotene.     

Supercritical carbon dioxide extraction of astaxanthin and other carotenoids from the microalga Haematococcus pluvialis

Supercritical carbon dioxide extraction of astaxanthin and other carotenoids from Haematococcus pluvialis was carried out, for several experimental conditions, using a semi-continuous apparatus. The microalga was previously freeze-dried and ground with a ball mill. The effects of pressure (200 and 300 bar), temperature (40 and 60 °C), degree of crushing, as well as the use of ethanol as a co-solvent (10%) on the extraction efficiency were assessed. Organic solvent extractions, using acetone, were also carried out in a vortex, on ground cells mixed with very small glass beads. Supercritical extraction from the completely crushed alga was compared with acetone and the highest recovery of carotenoids (92%) was obtained at the pressure of 300 bar and the temperature of 60 °C, using ethanol as a co-solvent.The extraction recovery increased with the pressure at 60 °C. On the other hand, the increase in temperature, at 300 bar, led to a slight improvement. The main carotenoid of Haematococcus pluvialis is the esterified astaxanthin (about 75%). Other carotenoids present are lutein, astaxanthin (free), β-carotene and canthaxanthin. All of them were recovered through supercritical fluid extraction with values higher than 90%, with the exception of canthaxanthin (about 85%), at a pressure of 300 bar and a temperature of 60 °C.     

Carotenoids in cassava: Comparison of open-column and HPLC methods of analysis

Cassava leaves and tubers from six cultivars were analysed for carotenoids by both open-column chromatography and high-performance liquid chromatography (HPLC). The major carotenoids of leaves were the non-vitamin A carotenoid lutein (86-290 mg kg^?1 fresh weight (FW)) and the pro-vitamin A carotenoid β-carotene (13-78 mg kg^?1 FW). Immature leaves contained less than mature leaves. Tubers contained 0.1-3 mg kg^?1 FW of β-carotene and 0.05-0.6 mg kg^?1 FW of lutein. Carotenoids present in small amounts included α-, γ-, and §-carotenes and β-cryptoxanthin and others were separated and partially identified. Yellow tubers contain much more β-carotene than white tubers and cassava is a good source of pro-vitamin A carotenoids compared with other root crops. Both methods gave similar results for β-carotene and lutein, but the open-column method did not allow separation and quantitation of carotenoids present in small amounts. HPLC separated many carotenoids, but most peaks could not be identified for lack of reference compounds. A great advantage of open-column chromatography for developing countries is its use of simple equipment and readily available reagents and solvents compared with HPLC, which requires complex equipment needing careful maintenance and expensive solvents. The open-column method is more labour intensive but this is not a constraint in developing countries.     

高效液相色谱法测定红小丑鱼中虾青素和叶黄素含量

为建立检测红小丑鱼皮肤和鳍组织中虾青素、叶黄素含量的高效液相色谱分析方法。样品经丙酮提取,甲基叔丁基醚萃取,纯水分离杂质,LC-NH₂净化,C₁₈色谱柱分离,流动相为甲醇:水(98:2,v/v),紫外检测波长475 nm,外标法定量。虾青素和叶黄素标准品0.02~1 μg/mL线性关系良好,虾青素相关系数为0.998,叶黄素相关系数为0.999,加标回收率分别为79.92%~90.32%和78.51%~97.60%,相对标准偏差分别为6.35%~7.07%和10.79%~13.41%。该方法前处理简单,能同时测定红小丑鱼组织中虾青素和叶黄素含量。