化学发光法对花生多肽的体外抗氧化活性和对DNA损伤的保护作用的研究

采用多个化学发光体系系统地研究了花生多肽的体外抗氧化活性及其对DNA损伤的保护作用。运用邻苯三酚-鲁米诺化学发光体系测定了花生多肽对超氧阴离子的清除作用,硫酸铜-邻菲哕啉-抗坏血酸-双氧水、硫酸亚铁-鲁米诺-双氧水和硫酸亚铁-鲁米诺3个体系测定了花生多肽对羟基自由基的清除作用,双氧水-鲁米诺体系测定了花生多肽对体外双氧水的清除作用,采用硫酸铜-邻菲哆啉-抗坏血酸-双氧水-脱氧核糖核酸测定了花生多肽对体外DNA损伤的保护作用。试验结果表明花生多肽具有较好的体外清除活性氧和保护DNA损伤的活性。花生多肽可作为开发抗氧化产品的新资源。     

儿茶素氧化产物的分离鉴定及其抗氧化活性研究

采用H2O2模拟氧化体系制备儿茶素氧化产物,经多级柱色谱分离得到一相对纯组分(tR为9.825min),HPLC-MS分析结果表明,所得级分主要含m/z471及m/z939两种产物,m/z471是EGCG分子上脱去2个氢原子,再加上一个氧原子形成,即[M 14-H]-,m/Z939为其二聚体,即[2M-3H]-。抗氧化活性测定结果表明,此氧化产物对不同体系(甲基紫体系、2-脱氧-D-核糖体系、水杨酸体系和邻苯三酚-鲁米诺化学发光体系)产生的羟自由基(·OH)和超氧阴离子自由基(O-2·)具有很强的清除作用,对前三个体系产生的羟自由基(·OH)的清除率分别为96.46%、84.8%、81.9%,在邻苯三酚-鲁米诺化学发光体系中对超氧阴离子自由基(O-2·)的清除率为97.0%,均高于同一体系下等浓度茶多酚。     

化学发光法测定樟树果红色素抗氧化活性的研究

采用化学发光法研究了樟树果实红色素(RPCF)的抗氧化活性及其对DNA损伤的保护作用。运用Pyrogallol-Luminol化学发光体系测定了RPCF对超氧阴离子的清除作用,CuSO4-Phen-VC-H2O2体系测定了RPCF对羟基自由基的清除作用,H2O2-Luminol体系测定了RPCF对双氧水的清除作用,CuSO4-Phen-VC-H2O2-DNA测定了RPCF对体外DNA损伤的保护作用。试验结果表明RPCF有较好的清除活性氧和保护DNA损伤的活性,其清除.O2-、.OH、H2O2能力及对DNA损伤的保护作用的半抑制浓度(IC50)分别为0.207,0.383,0.227和0.401 mg/mL。     

不同品种生姜粗黄酮抗氧化活性的比较

本文比较分析了品种对生姜粗黄酮抗氧化效果的影响,实验选用凤头姜老姜、凤头姜仔姜及黄爪姜老姜为原料,采用化学发光法测定了三种不同生姜粗黄酮在对超氧阴离子自由基、羟自由基、双氧水的清除作用和对DNA损伤的保护作用。结果发现,老姜黄酮的抗氧化效果优于仔姜,但在DNA损伤体系中,仔姜的保护效果要显著优于老姜。就品种而言,在双氧水体系中不同品种生姜粗黄酮的抗氧化效果差异不显著,其它三个体系差异性均达到了显著水平;在羟自由基、双氧水和DNA损伤保护作用体系中,凤头姜抗氧化效果优于黄爪姜的,但黄爪姜老姜粗黄酮对超氧阴离子自由基的清除作用要略优于凤头姜的,但在0.01水平上差异并不显著。综合比较发现品种和成熟期对生姜粗黄酮抗氧化效果均有影响。     

红景天提取物清除(O2-)·和·OH的体外实验研究

为了探讨红景天抗氧化保健作用以及为研究其抗氧化机理,用高效液相色谱法确定了红景天提取物中红景天苷的含量;采用邻苯三酚自氧化法、黄嘌呤-黄嘌呤氧化酶-鲁米诺法观察了红景天对超氧阴离子自由基(O_2~-)的清除作用;用水杨酸捕获法、丙二醛确定法、Phen-CuSO4-VC-H2O2-DNA法检测了红景天清除羟自由基·OH的效果。结果表明,红景天提取物能够减少邻苯三酚自氧化中间产物的生成、对体系中由黄嘌呤氧化酶催化黄嘌呤产生O_2~-·诱导鲁米诺发光有一定抑制作用。在水杨酸捕获法中红景天提取物半数清除体系中·OH的浓度(IC50)约为130μg/ml。用丙二醛法确定红景天提取物浓度为300μg/ml时,对体系中·OH抑制程度达到最大。在Phen-CuSO4-VC-H2O2-DNA体系中随着红景天提取物浓度的增加,DNA损伤产物发光峰较空白对照组明显后移且峰高明显降低。不同方法体外测定结果一致表明红景天提取物具有清除O_2~-·和·O H的作用。     

茄子皮红色素抗氧化活性研究

目的:研究茄子皮天然红色素体外抗氧化活性;方法:以抗坏血酸为对照,以Fenton反应产生羟自由基(·OH),肾上腺素自氧化反应产生超氧阴离子自由基(O-2·)为试验模型,采用紫外分光光度法和荧光分析法,测定茄子皮红色素对羟自由基、超氧阴离子自由基、过氧化氢的清除作用;结果:茄子皮红色素对上述自由基均有清除作用,时羟自由基和超氧阴离子自由基清除效果比抗坏血酸好,清除过氧化氢能力弱于抗坏血酸;结论:茄子皮红色素具有明显清除自由基的作用.     

杜仲翅果桃叶珊瑚甙抗氧化活性的研究

[目的]研究杜仲翅果桃叶珊瑚甙体外抗氧化活性。[方法]以抗坏血酸为对照,采用分光光度法测定杜仲翅果桃叶珊瑚甙对羟自由基、超氧阴离子自由基、过氧化氢自由基的清除作用。[结果]杜仲翅果桃叶珊瑚甙对上述自由基均有清除作用,过氧化氢自由基清除效果比抗坏血酸好,清除羟自由基和超氧阴离子自由基能力弱于抗坏血酸。[结论]杜仲翅果桃叶珊瑚甙具有一定的清除自由基的作用。     

红景天提取物清除O2-·和·OH的体外实验研究

为了探讨红景天抗氧化保健作用以及为研究其抗氧化机理,用高效液相色谱法确定了红景天提取物中红景天苷的含量;采用邻苯三酚自氧化法、黄嘌呤-黄嘌呤氧化酶-鲁米诺法观察了红景天对超氧阴离子自由基(O2-·)的清除作用;用水杨酸捕获法、丙二醛确定法、Phen-CuSO4-VC-H2O2-DNA法检测了红景天清除羟自由基·OH的效果.结果表明,红景天提取物能够减少邻苯三酚自氧化中间产物的生成、对体系中由黄嘌呤氧化酶催化黄嘌呤产生(O2-)·诱导鲁米诺发光有一定抑制作用.在水杨酸捕获法中红景天提取物半数清除体系中·OH的浓度(IC50)约为130μg/ml.用丙二醛法确定红景天提取物浓度为300μg/ml时,对体系中·OH抑制程度达到最大.在PhenCuSO4-VC-H2O2-DNA体系中随着红景天提取物浓度的增加,DNA损伤产物发光峰较空白对照组明显后移且峰高明显降低.不同方法体外测定结果一致表明红景天提取物具有清除(O2-)·和·OH的作用.     

三叉苦提取物抗氧化作用的研究

目的:研究三叉苦不同部位水提取物的抗氧化作用及其规律,为食品或药物抗氧化剂的筛选和应用提供理论依据。方法:应用化学发光法检测三叉苦的各部位水提取物对邻苯三酚 -CTMAB 体系产生的超氧阴离子自由基(O2·)、邻菲罗啉 - 抗坏血酸体系产生的羟自由基(·OH)及过氧化氢(H2O2)的清除作用。结果:三叉苦水提取物具有明显的清除超氧阴离子自由基、羟自由基和过氧化氢的作用,清除率与浓度之间存在着明显的量效关系。结论:证明了三叉苦各部位的水提取物均是一种有效的抗氧化剂。     

青钱柳提取物体外抗氧化活性研究

研究青钱柳不同溶剂提取物的体外抗氧化活性。测定了青钱柳不同溶剂提取物对DPPH自由基的清除作用,用化学发光法测定对超氧阴离子自由基(O·2)和羟自由基(·OH)的清除能力,并用烘箱储藏法测定对油脂的抗氧化活性。实验结果表明,醇提物具有较强的清除自由基能力和抗油脂氧化活性。     

荔枝皮原花青素与VC、VE的协同抗氧化研究

选取荔枝皮原花青素(LPPC),采用DPPH自由基清除实验与等辐射分析法(Isobologram)相结合,探讨荔枝皮原花青素、VC、VE单独和复配后的抗氧化作用,结果显示：荔枝皮原花青素、VC和VE的IC50值分别为9.98、9.21、27.96mg/L,表明荔枝皮原花青素对DPPH自由基有明显的清除作用。Isobologram分析图中荔枝皮原花青素与VC、VE复配后的效应点都在相加线及95%可信限的下方,理论IC50add值与实验IC50mix值有显著性差异,并且相互作用指数都小于1,证实荔枝皮原花青素与VC,荔枝皮原花青素与VE之间存在协同抗氧化效应。     

蛋白基体系美拉德反应在荔枝皮原花青素作用下的抑制研究

本试验分别研究了在乳清蛋白-葡萄糖和牛血清白蛋白-葡萄糖模拟生理体系中,荔枝果皮原花青素(Litchi pericarp procyanidins,LPPC)对美拉德反应和晚期糖基化终末产物(advanced glycation end products,AGEs)的抑制作用,以荧光性AGEs的强度为表征依据。结果表明,在乳清蛋白-葡萄糖模拟体系中孵育35 d时,LPPC对AGEs的抑制效果最强,达60.21±1.34%。LPPC浓度为1 mg/m L时,相对抑制率最大可达85.33±9.02%(显著高于维生素C(Vc),p0.05)。在牛血清白蛋白-葡萄糖体系中,孵育35 d后LPPC对AGEs的相对抑制率最高达70.01±1.32%,且与LPPC浓度呈现正相关。当LPPC浓度为0.5 mg/m L时,抑制率最大为95.46±10.12%(显著高于氨基胍(AG),p0.05)。不同p H值下蛋白质的稳定性研究表明,乳清蛋白在与LPPC长期孵育后,其热稳定性明显降低,再次提示了LPPC对美拉德反应的抑制作用。LPPC可作为一种天然的食品基质的AGEs抑制剂深度开发。     

An enhanced preparation and purification of the major antioxidants baicalein and wogonin from Scutellariae radix

A simple, rapid and improved method was developed to isolate high-purity baicalein and wogonin from Scutellariae radix. The method involves endogenous baicalinase-catalysed hydrolysis (EBCH), partition, automated low-pressure preparative chromatography (LPPC) and recrystallisation without multiple and tedious column chromatography. This process was optimised for large scale production of baicalein and wogonin with high yields, low costs and process automation. The transformation ratio of baicalin and wogonoside reached 98.21% and 96.60% after EBCH, leading to an increase of 5.41-fold in baicalein and 3.89-fold in wogonin, compared to a raw sample without hydrolysis. The purity of final products was more than 98% after one-step LPPC and recrystallisation. The experimental results show that EBCH–LPPC is an effective method for preparing high purity antioxidants.     

原花青素抑制丙烯酰胺的动力学

对原花青素对丙烯酰胺形成-消除动力学的影响进行研究。在荔枝原花青素（litchi pericarp procyanidin, LPPC）和莲房原花青素（lotus seedpod procyanidin,LSPC）对丙烯酰胺最大抑制率添加条件的基础上,分别采用 Logistic-生长曲线模型、Logistic-Fermi动力学模型和Logistic-指数动力学模型描述丙烯酰胺的形成-消除动力学过 程,最终优化选择Logistic-指数动力学模型为描述对象。实验结果表明：两种原花青素对丙烯酰胺形成过程具有显 著性影响,且LSPC的作用大于LPPC,在反应后期,两种原花青素对丙烯酰胺抑制均无显著性影响。     

蓝莓冻果多酚粗提物体外抗氧化活性的研究

选用过氧化氢体系、羟基自由基体系、超氧阴离子自由基体系、亚硝酸盐体系、还原能力对蓝莓冻果多酚粗提物进行抗氧化活性的测定,并与VC做对比.在试验浓度范围内(0.5～10 mg/mL),蓝莓冻果的多酚粗提物对几种体系有不同程度的抗氧化作用,清除过氧化氢能力较强,清除羟基自由基和亚硝酸盐的能力明显弱于VC,还原能力及清除超氧阴离子能力与VC较为接近.     

Initiation of lipid peroxidation in biological systems

The direct oxidation of PUFA by triplet oxygen is spin forbidden. The data reviewed indicate that lipid peroxidation is initiated by nonenzymatic and enzymatic reactions. One of the first steps in the initiation of lipid peroxidation in animal tissues is by the generation of a superoxide radical (see Figure 16), or its protonated molecule, the perhydroxyl radical. The latter could directly initiate PUFA peroxidation. Hydrogen peroxide which is produced by superoxide dismutation or by direct enzymatic production (amine oxidase, glucose oxidase, etc.) has a very crucial role in the initiation of lipid peroxidation. Hydrogen peroxide reduction by reduced transition metal generates hydroxyl radicals which oxidize every biological molecule. Hydrogen peroxide also activates myoglobin, hemoglobin, and other heme proteins to a compound containing iron at a higher oxidation state, Fe(IV) or Fe(V), which initiates lipid peroxidation even on membranes. Complexed iron could also be activated by O2- or by H2O2 to ferryl iron compound, which is supposed to initiate PUFA peroxidation. The presence of hydrogen peroxide, especially hydroperoxides, activates enzymes such as cyclooxygenase and lipoxygenase. These enzymes produce hydroperoxides and other physiological active compounds known as eicosanoids. Lipid peroxidation could also be initiated by other free radicals. The control of superoxide and perhydroxyl radical is done by SOD (a) (see Figure 16). Hydrogen peroxide is controlled in tissues by glutathione-peroxidase, which also affects the level of hydroperoxides (b). Hydrogen peroxide is decomposed also by catalase (b). Caeruloplasmin in extracellular fluids prevents the formation of free reduced iron ions which could decompose hydrogen peroxide to hydroxyl radical (c). Hydroxyl radical attacks on target lipid molecules could be prevented by hydroxyl radical scavengers, such as mannitol, glucose, and formate (d). Reduced compounds and antioxidants (ascorbic acid, alpha-tocopherol, polyphenols, etc.) (e) prevent initiation of lipid peroxidation by activated heme proteins, ferryl ion, and cyclo- and lipoxygenase. In addition, cyclooxygenase is inhibited by aspirin and nonsteroid drugs, such as indomethacin (f). The classical soybean lipoxygenase inhibitors are antioxidants, such as nordihydroguaiaretic acid (NDGA) and others, and the substrate analog 5,8,11,14 eicosatetraynoic acid (ETYA), which also inhibit cyclooxygenase (g). In food, lipoxygenase is inhibited by blanching. Initiation of lipid peroxidation was derived also by free radicals, such as NO2. or CCl3OO. This process could be controlled by antioxidants (e).(ABSTRACT TRUNCATED AT 400 WORDS)     

酶法水解脱脂米糠蛋白抗氧化性质研究

以脱脂米糠为原料,采用碱性蛋白酶、酸性蛋白酶和中性蛋白酶酶解制备米糠蛋白,并将制备酶解液与抗坏血酸在超氧阴离子自由基(O2-.)清除率、羟自由基(.OH)清除率、H2O2清除能力及还原能力等方面进行比较,研究米糠蛋白抗氧化活性。结果表明,米糠蛋白具有较强抗氧化活性,虽效果不如抗坏血酸;但对超氧阴离子自由基(O2-.)(最高为98.41%)、羟自由基(.OH)(最高达97.04%)及H2O2均有不同程度清除作用,并具有一定还原能力;且抗氧化能力与添加量存在一定量效关系,其中还出现有促氧化特殊现象。     

超声-微波协同法提取荔枝皮原花青素及其高聚体组成分析

为提高荔枝皮中原花青素的得率和进一步阐明其高聚体组成,本文以糯米糍荔枝为原料,采用超声-微波协同法提取荔枝皮原花青素(litchi pericarp proanthocyanidins,LPC),并运用响应面法优化其工艺条件,将分离所得的高聚原花青素(litchi pericarp polymeric proanthocyanidins,LPPC)用薄层层析色谱(TLC)初步定性,然后运用基质辅助激光解析电离-飞行时间质谱(MALDI-TOF-MS)分析其组成。结果表明,最佳提取工艺条件为:微波功率307W,时间17min,温度46℃,料液比1∶31g/mL,额定超声频率50kHz,优化后得率为8.10%;LPPC以四至六聚体为主,检测到最高聚体为十二聚体,没食子酸酰化程度低且富含A型键,由检测结果可见,各聚合度LPPC至少含有一种含A型键的结构。     

Activated oxygen species and oxidation of food constituents.

Activated oxygen species which may be important in initiating oxidative changes in foods include singlet oxygen, hydroxyl radical, ozone, superoxide anion (perhydroxyl radical at low pH), and hydrogen peroxide. Chemical and enzymic reactions known to occur in biological materials can generate singlet oxygen, hydroxyl radical, superoxide anion, and hydrogen peroxide. Ozone is primarily a product of photoreactions in polluted air. Reactions involving singlet oxygen, hydroxyl radical, and ozone with food constituents can ultimately yield peroxides which decompose to initiate oxidative chain reactions. Superoxide anion and hydrogen peroxide are relatively inert toward organic molecules but can decompose to produce the more reactive singlet oxygen and hydroxyl radical. Inhibition of reactions initiated by reactive oxygen species in foods should be very important in preserving the oxidative stability of foods. The generation, detection, measurement, reaction, and inhibition of reactions of active oxygen species are surveyed in this review.     

Activated oxygen species and oxidation of food constituents

Activated oxygen species which may be important in initiating oxidative changes in foods include singlet oxygen, hydroxyl radical, ozone, superoxide anion (perhydroxyl radical at low pH), and hydrogen peroxide. Chemical and enzymic reactions known to occur in biological materials can generate singlet oxygen, hydroxyl radical, superoxide anion, and hydrogen peroxide. Ozone is primarily a product of photoreactions in polluted air. Reactions involving singlet oxygen, hydroxyl radical, and ozone with food constituents can ultimately yield peroxides which decompose to initiate oxidative chain reactions. Superoxide anion and hydrogen peroxide are relatively inert toward organic molecules but can decompose to produce the more reactive singlet oxygen and hydroxyl radical. Inhibition of reactions initiated by reactive oxygen species in foods should be very important in preserving the oxidative stability of foods. The generation, detection, measurement, reaction, and inhibition of reactions of active oxygen species are surveyed in this review.