诃子抑菌活性成分筛选和总鞣质含量测定新方法

为了建立一种HPLC检测抑菌活性成分诃子总鞣质含量的新方法,在HPLC条件:流动相为乙腈-0.05%磷酸溶液梯度洗脱,检测波长为270 nm,流速为1 m L/min,柱温为30℃,色谱柱为Sepax Sapphire C18(250 mm×4.6 mm,5μm)下利用诃子鞣质中含量最高的成分诃子酸对药材中总鞣质进行含量测定。诃子酸在相应浓度内与色谱峰面积呈良好线性关系,平均加样回收率在97.22%~101.38%之间,RSD为1.62%。研究结果表明,该方法与比色法相比操作更简单,结果更准确。利用HPLC法来测定抑菌成分诃子总鞣质含量,可以更准确地对诃子进行质量控制。     

基于多元统计分析的不同诃子属药材多指标成分研究

建立了诃子属药材的多指标成分含量测定方法,为诃子属药材的质量评价提供依据.测定44批不同诃子属药材中7个鞣质类成分含量,并结合方差分析、聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)评价诃子属3种不同药材的鞣质类成分差异.可见诃子属3种药材中7个成分含量存在明显差异,方差分析结果...     

拳参中没食子酸和总鞣质的含量测定

该研究采用高效液相色谱法和紫外分光光度法分别对拳参中的主要成份———没食子酸以及总鞣质进行含量测定。其中,用高效液相色谱法测定没食子酸的含量,HPLC 条件为 Hypersil GOLD Phenyl 柱(250 mm ×4.6 mm,5μm),甲醇－0.1％磷酸梯度洗脱,流速为1.0 mL?min-1,检测波长为276 nm。用紫外分光光度法测定总鞣质的含量,采用2010版《中国药典》一部附录 XB 中鞣质含量测定方法,用干酪素沉淀鞣质,通过干酪素沉淀前后鞣质的变化来确定样品中总鞣质的含量。结果表明：没食子酸在0.051~1.02μg 范围内呈现良好的线性关系,平均加样回收率分别为99.1％、100.3％、101.9％,RSD 分别为2.1％、0.8％、2.0％;总鞣质在2.09~10.48μg 范围内呈现良好线性关系,平均加样回收率分别为102.2％、100.2％、102.1％,RSD 分别为1.3％、1.4％、1.0％。利用上述方法,还测定了贵州贵阳和四川成都各3批样品中没食子酸和总鞣质的含量差异,发现成都样品中的没食子酸与总鞣质均大于贵阳拳参中的含量,初步分析导致这一结果的原因可能与拳参的生长海拔有关,而成都的海拔范围更适合拳参的生长。上述两种方法快速、简单、重现性好,可以作为拳参中没食子酸和总鞣质的含量测定方法,而该方法也是首次被用来同时测定拳参中没食子酸和总鞣质的含量,为今后的地方药典标准甚至《中国药典》标准的补充和完善提供了重要依据。     

三七块根紫色素的花色苷本质及其含量和总皂苷含量的正相关性

三七是中国云南省的"第一药材",云南文山三七是三七的道地药材。三七块根的横截面为黄白色至紫色。紫色块根约占研究块根总数的28 .21 %,其中柱鞘、内皮层、皮层或表皮为紫色。特征颜色反应和紫外——可见光谱表明:三七块根紫色素属于黄酮类化合物,可能含有酚性邻位二羟基,不含类胡萝卜素、查耳酮、噢哢、异黄酮、儿茶素。花色苷和/或其苷元花色素奠定了紫色块根着色的基础,其他的非红色的黄酮类化合物起共色素的作用。块根的平均花色苷含量和平均总皂苷含量均以纯紫色块根的为最高,其次是黄紫混合色块根的,纯黄色的最低。块根的花色苷含量差异达到极显著水平,但总皂苷含量差异却没有达到显著水平。每个块根都含有不同量的花色苷,随花色苷量的增加,块根的紫色一般逐渐明显。块根的花色苷含量与其总皂苷含量之间呈显著正相关,相关系数r=0 .355。本文可为三七块根颜色呈现的机理探索及其色素的分子结构鉴定提供参考。     

七子花不同器官次生代谢产物含量的分析

对七子花不同器官的黄酮、鞣质、生物碱、皂甙、木质素、绿原酸等6种次生代谢产物的含量进行测定与分析。结果显示：6种次生代谢产物总量和各成分在七子花不同营养器官中的含量均具有一定的差异,总含量以老根最高,但主要以木质素为主;黄酮、总皂甙和总生物碱的含量以叶片最高,分别为24.13、5.51、31.08 g·kg^-1;绿原酸在花中含量最高,其次是叶,分别为38.74、37.13 g·kg^-1;鞣质在幼根中含量最高,为9.33 g·kg^-1;木质素以老根为最高,是120.43 g·kg^-1。这6种次生代谢产物在不同器官中含量与它们的合成途径、运输方向和作用部位有一定的关系。     

夏蜡梅营养器官总鞣质含量的比较

对中国特有植物夏蜡梅营养器官的总鞣质含量进行测定,并分析了与环境因子之间的相关性。结果表明:(1)夏蜡梅各营养器官均含有总鞣质,但以叶片的含量最高,根次之,一年生枝、二年生枝、茎等器官的含量很低。(2)夏蜡梅叶片的含量阳坡植株显著高于阴坡,根的含量则反之。(3)夏蜡梅7个样地叶片的总鞣质含量在1.1066%～2.0060%之间,平均为1.6906%,其中临安5个样地含量较低,大雷山2个样地较高,差异显著。(4)通径分析显示,夏蜡梅叶片总鞣质含量的主要影响因子为土壤氮含量和C/N比。     

商品鹿鞭中氨基酸含量比较

采用氨基酸自动分析仪测定不同品种与产地的9份鹿鞭药材中氨基酸成分的种类及含量。这9份样品中均含17种氨基酸,其中人体必需氨基酸6种。17种氨基酸中以甘氨酸含量最高,必需氨基酸中亮氨酸含量最高;比较不同样品的含量,以辽宁西丰的梅花鹿鞭的总氨基酸含量最高,黑龙江梅花鹿鞭的必需氨基酸含量最高。结论为不同产地鹿鞭中氨基酸含量有一定差异,该方法可用于鹿鞭药材的质量评价。     

伏毛铁棒锤不同器官生物碱含量动态变化研究

采用紫外分光光度法,对宁夏六盘山地区不同海拔、不同月份、野生和人工栽培的伏毛铁棒锤总生物碱含量进行分析.结果显示:(1)总生物碱在伏毛铁棒锤各器官中均有分布,其中以块根中含量最高(9.07±0.92 mg/g),叶的含量(3.2±1.7 mg/g)最低.(2)块根中的总生物碱含量随生长发育期的变化而不同,其中:6～11月份块根中总生物碱含量随生长期延长而逐渐升高,10月份块根含量达最高(9.07±0.92 mg/g).(3)在同一地区,总生物碱含量随海拔高度的升高而增加,在海拔高度为2 800 m时块根中含量达到10.04±0.51 mg/g,其他器官中总生物碱的含量变化基本相似.(4)在相同海拔高度下,野生伏毛铁棒锤总生物碱含量略高于人工栽培的,但差异不显著(P>0.05).研究表明,人工栽培的伏毛铁棒锤块根可以替代野生种做为药用,且在六盘山区人工种植的块根最佳采收期为10月,在选择人工种植区应考虑选择较高的海拔地区.     

块根紫金牛总黄酮提取工艺优化和不同产地、部位的含量比较

本研究采用正交试验分析和分光光度法,建立了块根紫金牛总黄酮含量测定的方法并比较分析其不同产地及不同部位总黄酮的含量差异.结果表明超声波法提取块根紫金牛中总黄酮的最佳提取工艺为:乙醇浓度60%,料液比1:70,提取时间50 min,提取功率200 W,提取温度70℃.另外,不同产地块根紫金牛中总黄酮含量顺序为:靖西＞那坡＞德保＞扶绥＞龙州＞武鸣;同一产地块根紫金牛根、茎、叶间总黄酮含量的高低顺序大致为:叶＞根＞茎.且通过精密度、重复性、稳定性和回收率实验来检测最佳提取工艺条件,结果显示该方法操作简便,结果准确,重复性好,RSD为0.788%,稳定性和回收率较高,平均回收率达96.20%.证明该方法可用于比较不同产地块根紫金牛药材中总黄酮的含量差异,对不同产地块根紫金牛的药材质量分析具有一定的指导作用.     

不同产地大血藤次生代谢产物含量比较

对不同产地大血藤的不同营养器官的总鞣质、总生物碱、皂苷、木质素、绿原酸、总黄酮共6种次生代谢产物的含量进行测定与比较。结果显示：6种次生代谢产物在不同营养器官中的含量具有一定的差异,总含量以叶最高,老茎次之,幼茎最小。6种次生代谢产物除木质素茎含量较高外,其余5种均以叶片的含量最高,差异具有显著性。不同产地大血藤的6种次生代谢产物的含量具有显著性差异。通过逐步回归分析和通径分析,得知大血藤叶片次生代谢产物与土壤生态因子的关系密切。聚类分析显示庆元百山祖及天台的大雷山和天台山的大血藤叶片的次生代谢产物含量较高。     

紫茎泽兰生长发育过程中糖、激素、单宁、黄酮的变化

采用高效液相和分光光度法对紫茎泽兰不同发育阶段（种子,芽,幼苗和成株）糖、激素、单宁及总黄酮含量差异进行了研究。结果表明：不同糖类的变化。紫茎泽兰的葡萄糖含量以芽最高,为6.93%;而种子（0.16%）、幼苗（1.27%）、成株(1.07%)均明显低于芽阶段;同样,紫茎泽兰中蔗糖含量芽较高,为2.68%,而种子（0.32%）、幼苗（0.52%）、成株(0.09%)均明显低于芽阶段。果糖含量的变化,最大值也是在芽阶段（3.28%）,其次为种子阶段（2.56%）,而种子和成株阶段均很低（0.14%和0.55%）。测定中,海藻糖、棉子糖、甘露糖的含量差别很大,仅种子阶段测定出3种糖的含量,结果表明差别不大,依次为0.04%、0.04%、0.03%。激素类的变化。在不同发育阶段,紫茎泽兰芽阶段吲哚乙酸（IAA）含量较高,达到14.20μg·g^-1 Dw ,比种子阶段高1.25μg·g^-1 Dw,是幼苗的1.63倍,成株阶段的5.55倍。紫茎泽兰赤霉素（GA）含量在不同发育阶段存在着明显的差异,而且种子的GA含量（901.11μg·g^-1 Dw）明显高于芽、幼苗和成株阶段,而以芽和幼苗阶段含量最低,分别为97.35和84.29μg·g^-1 Dw,成株阶段略高于芽和幼苗阶段,为280.50μg·g^-1 Dw。单宁类的变化：紫茎泽兰单宁变化差异明显,紫茎泽兰种子中单宁含量为1.98%,明显高于芽、幼苗和成株阶段（<0.30%）;总黄酮类的变化：紫茎泽兰总黄酮含量在发育过程中呈不断上升趋势。紫茎泽兰种子中总黄酮含量很微弱,为0.04%,而芽、幼苗和成株阶段的紫茎泽兰总黄酮含量均明显高于种子阶段,并且这3个阶段之间含量差别不大,分别为（0.60%、0.64%、0.62%）。这些结果说明,糖、激素、单宁及总黄酮含量与植物生长发育的基本规律和代谢过程的调控密切相关。     

桉树枝瘿姬小蜂危害对桉树次生代谢产物含量的影响

通过对24个桉树品系遭受桉树枝瘿姬小蜂危害前后次生物质含量单宁和类黄酮含量动态变化研究,结果表明桉树品系遭受枝瘿姬小蜂危害后,叶片内类黄酮含量明显上升,高感品系类黄酮含量和增长变幅明显低于其他抗性类型的品系;而桉树叶片内单宁含量的动态变化规律表现为：高抗品系单宁含量明显高于高感品系,但随着进入小蜂危害期,其叶片内单宁含量出现一定程度下降,随后升高。次生物质的动态变化在不同抗性品系间差异显著,表明桉树叶片内单宁和类黄酮含量与抗枝瘿姬小蜂有显著关系,可作为检测桉树对枝瘿姬小蜂抗性的指标。     

The effects of irradiance on the production of phenolic compounds and condensed tannins in Larix gmelinii needles

Needles of Larix gmelinii seedlings grown under different irradiances (100, 52, and 26 % of natural sunlight) were collected from June to August 2009. The content of phenolic compounds and condensed tannins in needles were strongly affected by different irradiances. The highest content of phenolic acids occurred under the lowest irradiance. Chlorogenic acid and syringic acid were detected only under the shade. In contrast, the needles under full irradiance showed the highest content of condensed tannins.     

A simple, integrative assay to quantify nutritional quality of browses for herbivores

Many regard the concentrations of nitrogen (N), tannins and plant cell wall constituents (fibre) as key indicators of food quality and habitat suitability for browsing herbivores; yet there is no method for measuring their combined effects. We have developed a simple in vitro assay for measuring the effects of tannins and fibre on N availability in browse. We determined the effects of tannins by measuring the polyethylene glycol (PEG)-binding capacity (PEG-BC) of Eucalyptus leaf samples, followed by a two-stage in vitro digestion with pepsin and cellulase to determine the digestibility of dry matter and N. There was a significant relationship between concentrations of digestible N and the PEG-BC of the leaves. Furthermore, adding PEG significantly improved the digestibility of N. Our results concur with in vivo observations from several mammalian species. This suggests that our method is effective for measuring the nutritional quality of browse and the benefits of adding PEG, providing some index of the detrimental effects of tannins. We further simplified the assay by removing the PEG step, allowing us to quickly analyse samples in bulk. Nevertheless, this simplified method is still not practical for analysing the many samples necessary to compare the nutritional values of different tracts of forest. We used near-infrared reflectance spectroscopy to produce calibration equations and predicted total and digestible N in 322 trees at eleven sites. Both within and between sites, we found a wide variation in concentrations of digestible N but a much lower variation in total N, with either no relationship or poor relationships between the two measures. This confirms the variability in the nutritional quality of eucalypt forests, which may explain the distribution and abundance of mammalian herbivores. Thus, our assay provides a useful tool for understanding how food resources influence herbivore populations at different scales. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

毛百合根尖染色体Giemsa C-带分析

本研究利用Giemsa C-带方法对毛百合(Lilium dahuricum Ker-Gawl)根尖染色体进行了分析。研究结果表明毛百合试管苗的染色体倍性变异丰富,染色体倍性变异包括二倍体(2n=2×=24)、三倍体(2n=3×=36)、四倍体(2n=4×=48)到六倍体(2n=6×=72)。对二倍体毛百合的C-带结果进行分析,其带型公式为:2n=2×=24=2CI++2CI+T+T++6I+2I++2I++2I+T++2I+T++2I+T++2T++2T+。每条染色体上都显示出显著的特征带,而且带纹的深浅差异明显。强带主要集中在长短臂上。因此,GiemsaC-带方法可以将毛百合(L.dahuricum)的每条染色体区分开。     

构树叶性状分化的生态功能初步研究

叶性状分化在自然界中较为普遍,不同的叶性状特征与植物对资源获得及利用效率密切关联,反映了植物适应特定环境所形成的生存对策。叶性状分化的生态功能一直以来备受生态学家和进化生物学家的广泛关注。自然界构树(Broussonetia papyifera)在个体发育过程中出现全缘叶和裂缺叶的分化,但其生态功能尚不清楚,推测两者的叶型分化是构树对虫害规避的结果。为了探讨构树叶性状分化对应的可能生态功能,该研究采用野外监测和室内分析的方法,对构树全缘叶和裂缺叶的虫害发生率、叶面积、与抗虫有关的酚类物质(总酚、缩合单宁、黄酮)含量进行了比较。结果表明:(1)相对于裂缺叶,全缘叶虫害发生率显著增加,全缘叶虫害发生率是裂缺叶的两倍。(2)自然条件下,全缘叶叶面积显著高于裂缺叶,增加了约44个百分点。(3)自然条件下,裂缺叶中总酚、缩合单宁、黄酮含量均显著高于全缘叶,分别提高了6.0%、4.2%和16.2%。(4)除黄酮外,虫害处理下裂缺叶中总酚、缩合单宁含量显著高于全缘叶,均提高了约5.0%。(5)人为移除部分叶片,裂缺叶中总酚、缩合单宁、黄酮含量均显著高于全缘叶,分别提高8.0%、1.6%和25.4%。这说明构树全缘叶和裂缺叶中酚类物质含量对外来损伤响应不一致,裂缺叶虫害发生率较全缘叶低可能由于两种类型叶片中酚类物质含量存在差异所引起。     

梨形四膜虫衰老过程中DNA含量的变化

应用一种新型的细胞核内DNA含量测定方法──图像分析法,测定真核细胞梨形四膜虫衰老过程中DNA含量的变化.根据Beer-Lambert定律,以细胞核在不同生长期内的积分光密度的水平表示核内DNA含量的变化.该方法具有测量速度快,重复性好,操作简单,结果可靠等优点.实验结果表明:四膜虫在进入对数生长期时,DNA含量逐渐达到高峰,随着细胞逐渐老化,细胞分裂次数及核内DNA含量逐步减少.     

The dynamics of phenolic compounds and soluble sugars in the leaves of the silver birch (<Emphasis Type="Italic">Betula pendula</Emphasis> Roth) after defoliation and their significance in entomological plant resistance

The dynamics of phenolic compounds, flavonols, catechines, tannins, and soluble sugars in the leaves of the silver birch Betula pendula Roth after strong (75%) and total (100%) artificial defoliation was studied. It was shown that the flavonol content in the leaves did not change after strong and total defoliation, while the amount of tannins did not change during the first 15 days but increased later on 1 and 2 years after 75% and 100% defoliation. The catechine content did not change during the first 15 days and increased later on 1 year after strong and total defoliation; however, it returned to the level of control plants 2 years after both types of defoliations. The amount of soluble sugars in the leaves increased 2 days after 75% defoliation; however, their content conformed to that in control plants after 10 days and it remained later 1 and 2 years after the damage. The amount of soluble sugars in the leaves also did not change 1 year after 100% defoliation.     

A critical analysis of techniques for measuring tannins in ecological studies

Summary A series of seventeen plant extracts rich in phenolic materials, including condensed and hydrolysable tannins, have been subjected to a series of chemical analyses in an attempt to gather ecologically significant information about their structure. Procedures investigated were (i) the Folin-Denis and Hagerman and Butler methods for quantifying total phenolics, (ii) the vanillin and proanthocyanidin methods for quantifying condensed tannins, (iii) the iodate and nitrous acid methods for hydrolysable tannins. It was found that the techniques for total phenolics correlated well, the Hagerman and Butler method giving higher estimates where solutions were particularly phenol rich. By contrast there was considerable discrepancy between the methods examined for condensed tannins. This is probably due primarily to the very different chemical reactions that form the basis of these procedures and also to the fact that the extract dependent products of the proanthocyanidin method vary in their A ₁ ¹ values. During the study of condensed tannins methods for estimating their contribution to total phenolics and for measuring their average polymer length were examined. In both cases different procedures produced very variable results. Available methods for hydrolysable tannins were found not to be generally applicable across all extracts thought to contain this type of tannin on the basis of chromatographic analysis. An attempt to produce a quantitative spectrophotometric assay for hydrolysable tannins based on changes in reactivity to ferric chloride due to hydrolysis is described. This proved to be of limited sensitivity but may have some merit for estimating levels in hydrolysable tannins in phenol-rich plant extracts that also contain condensed tannins. It is concluded that whilst the overall level of phenolics in extracts can be estimated with some confidence the information imparted by more specific assays is very dependent on the procedures employed, particularly when dealing with extracts from taxonomically highly diverse sources.     

Effect of tannic acid on Lactobacillus hilgardii analysed by a proteomic approach

AIMS: A contribution towards the elucidation of the mechanisms of tannins on bacteria growth inhibition, with particular focus on the interaction between tannins and bacterial proteins. METHODS AND RESULTS: The interaction between tannic acid (TA) and Lactobacillus hilgardii, a wine spoilage bacterium, was investigated by a combination of physiologic and proteomic approaches. Growing tests were performed on medium supplemented with TA at concentrations ranging from 100 to 1000 mg l(-1) demonstrating the inhibitory effect of TA on the growth rate. Total proteins extracted from cells unexposed and exposed to TA were then analysed by 2D-electrophoresis and significant quantitative variations with a marked decrease of protein intensity upon TA exposure were observed. Most of the proteins, identified by ESI tandem Mass Spectrometry, were metabolic enzymes of different pathways, located in cytoplasm and membrane. CONCLUSIONS: The effects of TA on cells are deduced by the involvement of metabolic enzymes, and functional proteins on the tannin-protein interaction. These results might be related to the altered functions of the cell metabolism. SIGNIFICANCE AND IMPACT OF THE STUDY: The possible role of tannins in the inhibition of the bacterial survival and growth in a natural environment such as wine. A similar approach could be applied for evaluating the effects of tannins on food borne and pathogenic bacteria.