蛹虫草无性型菌丝体提取液体外抗氧化活性研究

分别测定了蛹虫草无性型菌丝体不同溶剂提取液在不同抗氧化模型中的抗氧化作用。结果表明,各种提取液对二苯基苦味酰基苯肼自由基(·DPPH)和羟自由基(·OH)均有显著的清除作用:在50mg/mL时,去离子水、70%乙醇和70%丙酮提取物对DPPH自由基的清除率分别为89.2%、83.6%和75.9%;70%乙醇提取物在30mg/mL时对·OH自由基的清除率达到100%;在50mg/mL时70%丙酮和去离子水提取物的·OH自由基清除率分别为95.6%和89.6%。在一定浓度下,各提取液对邻苯三酚自氧化也均有抑制作用;不同溶剂提取物的还原能力强弱为:70%乙醇>去离子水>70%丙酮。各种提取物的抗氧化组分不同,提取物浓度与抗氧化性成正相关。     

蝉拟青霉代谢产物清除DPPH自由基和抗真菌活性的研究

用二苯代苦味肼基自由基(DPPH)-TLC法和酶标仪法对一株蝉拟青霉P3菌丝体和发酵液甲醇提取物的清除自由基活性进行了定性和定量测定,发现两种提取物具有较强的清除自由基活性,在浓度为5.0mg/mL,于37℃下保温10min时,两种样品对0.4mg/mL的DPPH自由基的清除率分别可达55.52%和74.86%.以一种黑曲霉菌为指示菌,采用薄层色谱法对蝉拟青霉菌丝体和发酵液甲醇提取物进行抑菌活性试验,实验中根据抑菌圈大小判定代谢物抑菌活性的大小.同时,以致病菌白色假丝酵母菌为指示菌,采用牛津杯法对提取物进行进一步抑菌活性验证,结果表明,菌丝体和发酵液提取物样品浓度为5.0mg/mL时,其抑菌圈直径分别可达11.23mm和21.42mm.     

花脸香蘑菌丝体提取物的体外抗氧化活性

分别测定花脸香蘑Lepista sordida发酵菌丝体不同浓度乙醇溶剂提取液在4种抗氧化模型中的抗氧化作用。结果表明, 各种菌丝体提取液对二苯基苦味酰基苯肼自由基(·DPPH)和羟自由基(·OH)均有显著的清除作用, 其清除作用随着乙醇溶剂浓度的提高而降低: 在3 g/L菌丝生药浓度时, 去离子水提取物对DPPH自由基和羟自由基的清除率分别为90.55%和81.9%; 在6 g/L菌丝生药浓度时, 50%乙醇和70%乙醇提取物对DPPH自由基的清除率分别为53.7%和45.2%, 对羟自由基的清除率分别为79.4%和78.4%。各菌丝提取物也具有极强的抗脂质过氧化能力, 在5 g/L菌丝生药浓度64 h内, 水提物、50%乙醇和70%乙醇提取物对亚油酸过氧化物的抑制率分别为100%、96%和89.2%。在一定浓度下, 各提取液对邻苯三酚自氧化也均有抑制作用。     

鸡骨草黄酮体外抗活性氧自由基作用的研究

对鸡骨草进行黄酮提取及抗活性氧自由基作用的研究。以70%乙醇为溶剂,超声波提取法提取鸡骨草中的黄酮;采用比色法测定鸡骨草黄酮对DPPH·和O-·2两种自由基的清除作用以及还原能力。鸡骨草黄酮对DPPH·和O-·2两种自由基都有明显的清除作用,清除能力为DPPH·O-·2;对Fe3+也具有较强的还原能力。清除率与浓度存在明显的量效关系,当黄酮的浓度为0.306 mg/mL时,其对O-·2清除率为12.57%;当黄酮的浓度为0.228 mg/mL时,其对DPPH·清除率为50%。对自由基的清除能力总体强弱是:DPPH·O-·2;还原Fe3+能力随黄酮的浓度增加而增强。     

柿子渣中多酚的提取工艺及其抗氧化性研究

采用单因素试验设计,研究了料液比、提取温度、乙醇浓度、提取时间4个因素对柿子渣中总多酚提取效果的影响,运用正交试验设计对提取工艺进行了优化,并采用DPPH法检测了柿子渣多酚提取液清除自由基的能力.结果表明:(1)提取温度和提取时间对总多酚含量影响显著,乙醇浓度和料液比对总多酚含量影响不显著;柿子渣总多酚提取的最佳工艺为:料液比20:1(mL·g-1),提取温度90℃,乙醇浓度40%,提取时间4 h.(2)验证试验结果表明,优化条件下总多酚的提取量可达到21.402 mg·g-1,高于正交试验中任何一组.(3)抗氧化活性试验表明,当柿子渣多酚提取液质量浓度为0.24 mg·mL-1时,对DPPH自由基的清除率可达95.4%,与对照Vc抗氧化能力无显著差异,表明在该优化工艺下提取的柿子渣多酚具有较强的抗氧化活性.     

瓦宁木层孔菌中多酚和黄酮类成分分离及清除自由基活性的研究

采用硅胶和反相C18柱层析方法,首次从瓦宁木层孔菌中分离得到了5个化合物,运用NMR波谱法分析和鉴定为樱花亭、7-甲氧基二氢莰非素、二氢莰非素、4-(3,4-二羟苯基)-3-丁烯-2-酮、hispolon。并通过建立体外二苯基苦味酰基苯肼自由基（·DPPH）、超氧阴离子自由基（·O2?）以及羟自由基（·OH）发生体系,研究了5个化合物对·DPPH、·OH和·O2?的清除作用。结果表明当浓度达到100μg/mL时,化合物4-(3,4-二羟苯基)-3-丁烯-2-酮和hispolon对·DPPH清除率分别为92%和93%,对·OH的清除率分别为90%和95%,而对·O2?的清除率分别为70%和77%,略低于清除·DPPH和·OH的能力;二氢莰非素对·O2?自由基的清除率为39%,强于清除·OH和·DPPH的能力;而樱花亭和7-甲氧基二氢莰非素对3种自由基的清除率均低于30%。2个多酚类化合物清除自由基的能力均强于3个黄酮类化合物。5个化合物清除自由基能力均表现出一定的浓度依赖性。     

低聚壳聚糖衍生物的制备及其抗氧化性能

低聚壳聚糖(COS)经化学改性得到N-苯亚甲基壳聚糖(NBCOS)和O-2'-羟丙基三甲基氯化铵壳聚糖(OHCOS),对其结构进行表征.考察了COS及其衍生物对超氧阴离子O2¨、羟基自由基·OH、DPPH自由基的清除活性以及还原能力.结果表明:当浓度为10 mg/mL时,COS和OHCOS对O2¨的清除率分别为89.6%和85.5%,而同样浓度时NBCOS的清除率仅有6.9%;它们清除·OH和DPPH的活性大小顺序为COS>OHCOS>NBCOS,而还原能力大小顺序为COS>NBCOS>OHCOS.     

夏枯草色素的抗氧化活性研究

目的:对夏枯草色素的抗氧化作用进行研究.方法:以蒸馏水为溶剂,从夏枯草中提取色素;采用比色法测定夏枯草色素对DPPH·、O2-·和·OH 3种自由基的清除作用以及还原能力.结果:夏枯草色素对三种自由基都有明显的清除作用,清除能力为DPPH·＞·OH＞ O2-·;对Fe3+也具有较强的还原能力.结论:清除率与浓度存在明显的量效关系,当色素浓度为0.13 mg/mL时,其清除率O2-·为4.1％、DPPH·为28.3％、·OH为17.9％;还原Fe3+能力随色素的浓度增加而增强.     

野菊花60%乙醇提取物的酚类成分组成及其清除自由基和防霉变能力分析

对野菊[Dendranthema indicum (L. ) Des Moul. ]花的60%乙醇提取物中酚类成分的组成与含量进行了分析,并研究了该提取物对· O2-、· OH和DPPH自由基的清除能力以及防牛奶霉变的能力.结果表明,野菊花60%乙醇提取物主要含有黄酮类成分和一定量的酚酸类成分;酚酸类成分中绿原酸和咖啡酸的含量分别为20.74 mg·g-1和1 420.57 μg·g-1;总黄酮含量为490.50 mg·g-1,其中槲皮素、木犀草素、芹菜素、刺槐素和蒙花苷的含量分别为31.05、22.67、23.09、30.71和107.23 mg·g-1,分别占总黄酮含量的6.33%、4.62%、4.71%、6.26%和21.86%.该提取物对· O2-、· OH和DPPH自由基均有一定的清除能力,在实验设置的浓度范围内,对3种自由基的清除率与该提取物浓度基本呈正相关关系,回归方程分别为y=0.273x+38.540、y=1.208x+2.761和y=2.032x+45.330,IC50分别为41.98 μg·mL-1、39.11 mg·mL-1和2.30 mg·mL-1,其中对DPPH自由基的清除能力最强,且其IC50略高于VC.在牛奶中添加一定量的野菊花60%乙醇提取物(质量体积分数0.012 5%～0.2%),牛奶中的各级霉斑数量、病情指数以及霉菌孢子数均低于空白处理组,且该提取物的这种防霉变能力与其浓度呈一定的正相关关系;其中0.2%的野菊花60%乙醇提取物的防霉变能力高于质量体积分数0.01%山梨酸钾.实验结果显示,野菊花60%乙醇提取物具有较高的抗氧化能力和显著的防霉变能力,可作为食品防腐剂进行开发利用.     

超声波协同提取鱼腥草黄酮及其抗氧化性

研究了超声波协同乙醇提取不同部位鱼腥草黄酮的最佳工艺条件,并测定黄酮提取物对DPPH.、羟基自由基和超氧自由基的抑制效果。结果表明:鱼腥草叶中黄酮含量最高,在固液比1∶30,85%乙醇浓度,65℃下提取40min,黄酮提取率为2.36%。鱼腥草黄酮对DPPH.自由基活性、.OH自由基和超氧O2-.自由基的清除率可达72.8%、70.8%和69.8%,表明鱼腥草黄酮是一种天然有效的自由基清除剂。     

炭疽病菌侵染对荔枝果实生理生化变化的影响

本研究测定了荔枝果实人工接种炭疽病菌后呼吸速率、乙烯释放量的变化和果皮氧化、过氧化作用以及与酚类代谢有关的几种酶活性的变化。结果表明,接种炭疽病菌的荔枝果实呼吸速率和乙烯释放量显著增高,果皮活性氧(O₂^·)产生速率和丙二醛(MDA)含量显著增加,超氧化物歧化酶(SOD)活性显著降低,过氧化物酶(POD)、多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)活性显著增高。说明炭疽病菌的侵染可导致荔枝果实呼吸速率和乙烯释放量的增高,加速荔枝果皮氧化和过氧化进程,并诱导荔枝果皮PPO、POD、PAL活性增高,是加速采收后荔枝果实衰老、褐变、腐烂的一个重要原因。     

红背叶提取物的体外抗氧化活性

采用DPPH法、FRAP法、ABTS法和清除羟基自由基法四种抗氧化活性方法,测定了红背叶不同溶剂萃取60%EtOH提取物得到的部位清除自由基的能力.结果表明,乙酸乙酯提取物的抗氧化能力最强,强于阳性对照VC和Trolox;其次是60%乙醇提取物,其抗氧化能力基本与阳性对照BHA相当.     

Optimized ultra-high-pressure-assisted extraction of procyanidins from lychee pericarp improves the antioxidant activity of extracts

To establish optimal ultra-high-pressure (UHP)-assisted extraction conditions for procyanidins from lychee pericarp, a response surface analysis method with four factors and three levels was adopted. The optimum conditions were as follows: 295 MPa pressure, 13 min pressure holding time, 16.0 mL/g liquid-to-solid ratio, and 70% ethanol concentration. Compared with conventional ethanol extraction and ultrasonic-assisted extraction methods, the yields of the total procyanidins, flavonoids, and phenolics extracted using the UHP process were significantly increased; consequently, the oxygen radical absorbance capacity and cellular antioxidant activity of UHP-assisted lychee pericarp extracts were substantially enhanced. LC-MS/MS and high-performance liquid chromatography quantification results for individual phenolic compounds revealed that the yield of procyanidin compounds, including epicatechin, procyanidin A2, and procyanidin B2, from lychee pericarp could be significantly improved by the UHP-assisted extraction process. This UHP-assisted extraction process is thus a practical method for the extraction of procyanidins from lychee pericarp.     

荔枝壳主要黄烷醇类物质分析

本研究采用乙醇溶液提取荔枝壳中的黄酮,并利用不同有机溶剂进行分级。通过反相液相色谱对主要单体成分P1,P2和P3分离纯化。经紫外可见光扫描分析表明为黄烷醇类物质。通过核磁共振波谱和质谱分析证明P1,P2和P3分别为表儿茶素,原花青素B2和原花青素B4。     

Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.     

荔枝壳多糖特性研究

多糖是荔枝壳的重要活性成分,本研究采用阴离子交换柱和凝胶过滤柱对荔枝壳多糖进行分离纯化。凝胶渗透色谱测定其分子量为14000 Dal。通过气相色谱测定其单糖组成为甘露糖、半乳糖和少量的阿拉伯糖,分子链由1,2键、1,3键和1,6键组成,不含1,4键。红外光谱分析表明甘露糖以β-D-甘露糖形式存在,不含羧基基团。     

Cloning and Expression Analysis of Litchi (Litchi Chinensis Sonn.) Polyphenol Oxidase Gene and Relationship with Postharvest Pericarp Browning

Polyphenol oxidase (PPO) plays a key role in the postharvest pericarp browning of litchi fruit, but its underlying mechanism remains unclear. In this study, we cloned the litchi PPO gene (LcPPO, {"type":"entrez-nucleotide","attrs":{"text":"JF926153","term_id":"350601663","term_text":"JF926153"}}JF926153), and described its expression patterns. The LcPPO cDNA sequence was 2120 bps in length with an open reading frame (ORF) of 1800 bps. The ORF encoded a polypeptide with 599 amino acid residues, sharing high similarities with other plant PPO. The DNA sequence of the ORF contained a 215-bp intron. After carrying out quantitative RT-PCR, we proved that the LcPPO expression was tissue-specific, exhibiting the highest level in the flower and leaf. In the pericarp of newly-harvested litchi fruits, the LcPPO expression level was relatively high compared with developing fruits. Regardless of the litchi cultivar and treatment conditions, the LcPPO expression level and the PPO activity in pericarp of postharvest fruits exhibited the similar variations. When the fruits were stored at room temperature without packaging, all the pericarp browning index, PPO activity and the LcPPO expression level of litchi pericarps were reaching the highest in Nandaowuhe (the most rapid browning cultivar), but the lowest in Ziniangxi (the slowest browning cultivar) within 2 d postharvest. Preserving the fruits of Feizixiao in 0.2-μm plastic bag at room temperature would decrease the rate of pericarp water loss, delay the pericarp browning, and also cause the reduction of the pericarp PPO activity and LcPPO expression level within 3 d postharvest. In addition, postharvest storage of Feizixiao fruit stored at 4°C delayed the pericarp browning while decreasing the pericarp PPO activity and LcPPO expression level within 2 d after harvest. Thus, we concluded that the up-regulation of LcPPO expression in pericarp at early stage of postharvest storage likely enhanced the PPO activity and further accelerated the postharvest pericarp browning of litchi fruit.     

采后荔枝果实中氧化和过氧化作用的变化

采后的荔枝（Litchi chinensts)果实的果皮和果汁中的抗坏血酸及果汁的谷胱甘肽含量逐渐下降,果皮的谷胱甘肽甘肽含量先增多（采后3天）接着减少,超氧化物歧化酶活性随采后时间加长而降低,膜脂过氧化产物丙二醛及过氧化物酶活性显著增高,采收7天后果皮中丙二醛含量增加2倍,过氧化物酶活性增高6－7倍,超氧物歧化酶活性只为原来的44％,内源清除活性氧能力的减弱与膜脂过氧化产物的积累表明,荔枝果实的衰老与活性氧的伤害有关,过氧化物酶活性增高可作为果实衰老的一个指标。