响应面优化伏毛铁棒锤总生物碱微波辅助提取工艺

目的：优化微波辅助提取伏毛铁棒锤总生物碱工艺，并比较不同产地伏毛铁棒锤中总生物碱含量。方法：以单因素试验为基础，结合响应面法，以总生物碱的含量为评价指标，考察温度、料液比、时间和体积分数对提取得率的影响。结果：最佳微波辅助提取工艺参数（条件）为：温度48℃，料液比1∶19(g/mL)，时间12 min，甲醇体积分数39%。结论：该提取方法稳定可靠，操作简便，可用于优化伏毛铁棒锤总生物碱提取工艺。比较不同地区伏毛铁棒锤中总生物碱含量，贵南县伏毛铁棒锤总生物碱含量最高，为0.290%。     

宁波地区野生换锦花鳞茎的主要成分含量测定

为了研究宁波地区野生换锦花鳞茎中主要成分的含量差异,以采自舟山大猫岛及宁波市北仑洋涨岙、奉化萧王庙、象山鹤浦、镇海招宝山、慈溪海黄山等6个地区的野生换锦花鳞茎为实验材料,以石蒜碱计总生物碱的含量,采用分光光度法测定其总黄酮、非淀粉粗多糖及总生物碱的含量;采用HPLC法测定其加兰他敏的含量。实验结果表明,6个地区的野生换锦花鳞茎中主要成分的含量存在一定的差异。不同地区的野生换锦花鳞茎中的总黄酮、总生物碱、加兰他敏的含量及非淀粉粗多糖的得率变化幅度分别为1.095~1.751 mg/g、2.691~5.559 mg/g、0.686~1.476 mg/g和11.590%~14.182%。其中采自象山鹤浦的总黄酮含量最高,为(1.751±0.116)mg/g;慈溪海黄山的非淀粉粗多糖得率最高,为(14.182±0.523)%;镇海招宝山的总生物碱含量最高,为(5.559±0.394)mg/g;北仑洋涨岙中加兰他敏含量最高,为(1.476±0.018)mg/g。同一地区样品中总生物碱与加兰他敏的含量呈一定的正相关,相关系数为0.601,但并不显著。本研究结果为换锦花人工栽培资源的选优及开发利用提供理论依据。     

野生地木耳中次生代谢产物含量测定与分析

测定了野生地木耳（Nostoc commune Vauch.）中次生代谢产物含量,并与4种常见食用菌藻黑木耳（Auricularia auricular-judae）、银耳（Tremella fuciformis）、紫菜（Porphyra）、海带（Laminaria japonica）进行比较分析,总酚、总黄酮和缩合单宁含量测定采用分光光度法,总生物碱含量测定采用高效液相色谱法（HPLC）。结果显示,野生地木耳中总酚含量为24.255 mg/g±1.631 mg/g,总黄酮含量为5.741 mg/g±0.239 mg/g,总生物碱含量为0.768 mg/g±0.073 mg/g,缩合单宁含量为0.069 mg/g±0.009 mg/g。4种食用菌藻中次生代谢产物含量范围为：总酚（5.520～62.326 mg/g）、总黄酮（0.847～7.010 mg/g）、总生物碱（0.408～4.132 mg/g）、缩合单宁（0.063～0.233 mg/g）;比较分析结果显示,野生地木耳次生代谢产物中的总酚和总黄酮物质含量较高,且总酚是主要次生代谢产物,缩合单宁与总生物碱含量较低;总酚含量约为黑木耳和银耳的2倍、紫菜的6倍;总黄酮含量约为黑木耳的7倍,银耳的3倍。     

滇龙胆不同部位裂环烯醚萜类物质含量比较分析

采用高效液相色谱法,对云南临沧云县滇龙胆不同部位龙胆苦苷、獐牙菜苦苷、当药苷含量进行分析.结果显示:(1)龙胆苦苷、獐牙菜苦苷、当药苷在滇龙胆根、茎、叶、花中均有分布;龙胆苦苷、獐牙菜苦苷在根部含量较高,分别为(35.88±5.89)mg/g和(1.35±0.33)mg/g;当药苷在花部含量最高,为(0.97±0.24)mg/g.(2)不同部位有效成分间具有显著(P<0.05)或极显著(P<0.01)的相关性.(3)相同海拔条件下,不同栽培方式下滇龙胆根部龙胆苦苷含量差异不显著(P>0.05),但茎部、叶部獐牙菜苦苷含量差异显著(P<0.05).     

人工栽培唐古特大黄中蒽醌含量水平的分析

采用醋酸镁比色法测定青海省不同海拔种植基地中栽培唐古特大黄的总蒽醌含量,并和野生大黄药材进行了比较。结果表明,在海拔较高地区栽培的二、三年生唐古特大黄中,其总蒽醌含量均明显高于较低海拔地区栽培的同龄唐古特大黄,3年生人工栽培大黄总蒽醌含量只有野生大黄药材的一半左右。     

洪湖湿地生态系统土壤有机碳及养分含量特征

洪湖湿地是长江中游地区重要湿地生态区域,在维持区域生态安全中发挥着重要作用.通过对洪湖湿地5种类型土壤进行有机碳及养分含量进行研究,为湿地生态系统地球化学循环及碳储量估算提供重要基础.结果表明,(1)洪湖土壤有机碳含量随土壤容重增加递减.(2)洪湖湿地土壤有机碳平均含量为:湖中淤泥＞洪泛平原湿地＞草本沼泽＞林地＞农田,其中湖底淤泥的有机碳平均含量6.74％,农田的有机碳平均含量为1.241％.此外,有机碳含量随土层的加深而减少.(3)湖底淤泥N的平均含量(4.623±0.535) g/kg、Ca的平均含量(26.262±4.201) g/kg,与洪泛平原湿地、草本沼泽及林地有显著性差异.农田P的平均含量(2.876±1.253) g/kg-、K的平均含量(7.205±0.159) g/kg,与湖底淤泥、洪泛平原湿地、草本沼泽及林地有显著性差异.(4)湖底淤泥中Cu的平均含量(40.19±3.04) mg/kg,Fe的平均含量(8560.90±80.98) mg/kg,Co的平均含量((29.66±0.67) mg/kg),Cu、Fe和Co的平均含量都显著小于洪泛平原湿地、农田及林地,Pb的平均含量(35.13±2.40) mg/kg,Mn的平均含量(749.65±54.07) mg/kg,这两个元素的含量均与农田及林地有显著性的差异.湖底淤泥中重金属元素的平均含量小于其余4种湿地类型土壤.     

农林复合系统滇龙胆茎叶化学计量特征研究

以农林复合系统种植的滇龙胆(Gentiana rigescens Franch.ex Hemsl.)为材料,采用高效液相色谱法建立不同栽培系统滇龙胆茎、叶的色谱指纹图谱,并测定其主要活性成分马钱苷酸、獐牙菜苦苷、龙胆苦苷和当药苷含量,研究不同栽培系统滇龙胆茎、叶化学计量特征。采用相关性分析、指纹图谱相似度分析、偏最小二乘判别分析(PLS-DA)、变量投影重要性准则(VIP)等方法进行化学数据分析。结果显示,滇龙胆主要活性成分马钱苷酸含量为(1.85±0.92)mg/g~(7.43±7.64)mg/g,獐牙菜苦苷含量为(1.03±0.17)mg/g~(1.58±0.50)mg/g,龙胆苦苷含量为(15.28±11.34)mg/g~(24.59±7.84)mg/g,当药苷含量为(4.10±1.64)mg/g~(31.67±22.70)mg/g,且叶片中4种活性成分的总含量高于茎;不同栽培系统中,与尼泊尔桤木间作的滇龙胆茎、叶活性成分总含量最高,而与核桃间作的滇龙胆茎、叶活性成分总含量最低。相关性分析显示,植株相同部位和不同部位间的环烯醚萜和裂环烯醚萜含量呈显著(P0.05)或极显著(P0.01)正相关。指纹图谱相似度分析表明,不同栽培系统滇龙胆茎指纹图谱相似度介于0.989~0.992之间、叶指纹图谱相似度为0.988~0.996,相同部位样品化学成分种类相似。PLS-DA分析结果表明,茎和叶片整体化学计量特征具有明显差异;单作及林药间作的样品被区分为不同类群,不同间作模式下滇龙胆茎、叶化学成分具显著差异,叶片高效液相色谱指纹图谱可用于区分不同栽培系统滇龙胆样品。本研究结果可为农林复合系统滇龙胆有效成分含量研究及滇龙胆资源的合理开发利用提供科学依据。     

人工培殖冬虫夏草D-甘露醇的含量测定

采用水提回流离心的方法,提取分离人工培殖冬虫夏草中D-甘露醇,用高碘酸钠与甘露醇反应产生黄色的3,5-二乙酰-1,4-脱氢二甲基吡啶,在波长412 nm处测定其最大吸收.甘露醇浓度在10～50 mg/L之间时,与吸光值(OD412)有良好线性关系,线性方程为C=79.172x-0.0713,相关系数r=0.9999.人工培殖冬虫夏草中D-甘露醇的含量分别为89.64 mg/g(2005年)、91.86 mg/g(2006年)、94.71 mg/g(2007年)、98.98 mg/g(2008年)、97.50 mg/g(2009年)、94.71 mg/g(野生)和96.84 mg/g(野生).人工培殖冬虫夏草与野生冬虫夏草D-甘露醇的含量基本相同.     

不同海拔云南黄连生物量和主要有效成分变化

研究了不同海拔(2 100～2 700 m)下,野生和人工栽培云南黄连的生物量、主要有效成分含量及产量.结果表明:野生云南黄连根茎和根生物量沿海拔梯度呈上升趋势,但无显著性差异(P>0.05);人工栽培云南黄连根茎生物量平均值在海拔2 600 m和2 700 m处分别为87.5 kg·hm^-2和97.0 kg·hm^-2,显著高于海拔2 300 m处(34.8 kg·hm^-2,P<0.05),且海拔2 300、2 600和2 700 m的人工栽培云南黄连根茎和根生物量均大于野生云南黄连,但无显著性差异(P>0.05). 野生云南黄连的根茎和根生物量均与全株生物量呈显著正相关. 野生云南黄连根茎和根小檗碱含量在海拔2 700 m处最高,分别为4.60%和1.93%; 根茎巴马汀和药根碱含量、根药根碱含量在海拔2 600～2 700 m处最高;根巴马汀含量在2 300 m处最高.人工云南黄连根茎和根小檗碱含量在海拔2 600 m处最高,分别为4.41%和1.90%; 根茎巴马汀含量,根小檗碱、巴马汀和药根碱含量在海拔2 600～2 700 m处最高;根茎药根碱含量在海拔2 300 m处最高.海拔2 600～2 700 m处野生云南黄连根茎和根中各有效成分产量显著高于海拔2 100和2 300 m处(P<0.05). 野生云南黄连分株的根茎生物量、根生物量、叶生物量、总生物量、高度和冠幅沿海拔梯度呈先升后降趋势.增大种植密度和加强人工管理可以提高云南黄连生物量和主要有效成分产量.     

短时高温胁迫对中华地鳖蛋白质和氨基酸含量的影响

土鳖虫体内富含蛋白质和氨基酸,是一种重要的药用昆虫。高温对昆虫生命活动产生重要影响。为探明短时高温胁迫对土鳖虫体内蛋白质和氨基酸含量的影响,本试验以中华地鳖Eupolyphaga sinensis Walker 8龄雌若虫为研究对象,通过人工设置29℃、33℃、37℃为高温处理组,以25℃为对照,测定2 h短时高温胁迫对中华地鳖存活率以及体内蛋白质和氨基酸含量的影响。结果表明:29℃、33℃和37℃短时高温处理对中华地鳖存活率无影响,蛋白质含量分别为76.57%±3.29%、79.60%±0.92%、77.00%±0.71%,与对照(84.50%±1.57%)相比均明显下降;短时高温处理后,29℃处理组土鳖虫总氨基酸含量为206.97±7.42 mg/g,6种人体必需的游离氨基酸苏氨酸8.23±0.32 mg/g,甲硫氨酸3.98±0.49 mg/g,异亮氨酸8.27±0.35 mg/g,亮氨酸16.38±0.86 mg/g,苯丙氨酸8.34±0.40 mg/g,以及赖氨酸12.56±0.64 mg/g的含量较对照和33℃、37℃高温处理组均显著升高,表明29℃短时高温处理可显著提高中华地鳖体内人体必需游离氨基酸含量。本研究是国内外对高温胁迫下药用昆虫体内活性成分的首次报道,研究结果将为进一步明确高温对土鳖虫的药用价值的影响,以及为土鳖虫人工养殖条件提供理论依据。     

旱生和湿生生境对蒲公英体内抗氧化物质的影响

选择旱生和湿生生境中生长的蒲公英(Taraxacum mongolicum Hand.-Mazz)分根、叶、花序分别测定抗氧化成分及总抗氧化能力,并比较了SOD同工酶谱的变化情况。结果表明不同生境选取的蒲公英叶片中抗氧化物质以SOD、POD、CAT等抗氧化酶和Vc等小分子为主,花序中可溶性糖、类黄酮和绿原酸含量最高;湿生蒲公英各器官尤其是根中的SOD、CAT、POD活性高于旱生蒲公英对应器官中的酶活性,旱生蒲公英各器官尤其是花序中的可溶性糖、类黄酮和绿原酸含量高于湿生蒲公英对应器官中的含量;旱生和湿生生境未诱导出新的SOD同工酶带;旱生和湿生蒲公英对应器官的总抗氧化能力接近。     

Tissue distribution and biosynthesis of 1,2-saturated pyrrolizidine alkaloids in Phalaenopsis hybrids (Orchidaceae)

Phalaenopsis hybrids contain two 1,2-saturated pyrrolizidine monoesters, T-phalaenopsine (necine base trachelanthamidine) and its stereoisomer Is-phalaenopsine (necine base isoretronecanol). T-Phalaenopsine is the major alkaloid accounting for more than 90% of total alkaloid. About equal amounts of alkaloid were genuinely present as free base and its N-oxide. The structures were confirmed by GC-MS. The quantitative distribution of phalaenopsine in various organs and tissues of vegetative rosette plants and flowering plants revealed alkaloid in all tissues. The highest concentrations were found in young and developing tissues (e.g., root tips and young leaves), peripheral tissues (e.g., of flower stalks) and reproductive organs (flower buds and flowers). Within flowers, parts that usually attract insect visitors (e.g., labellum with colorful crests as well as column and pollinia) show the highest alkaloid levels. Tracer feeding experiments with (14)C-labeled putrecine revealed that in rosette plants the aerial roots were the sites of phalaenopsine biosynthesis. However active biosynthesis was only observed in roots still attached to the plant but not in excised roots. There is a slow but substantial translocation of newly synthesized alkaloid from the roots to other plant organs. A long-term tracer experiment revealed that phalaenopsine shows neither turnover nor degradation. The results are discussed in the context of a polyphyletic molecular origin of the biosynthetic pathways of pyrrolizidine alkaloids in various scattered angiosperm taxa. The ecological role of the so called non-toxic 1,2-saturated pyrrolizidine alkaloids is discussed in comparison to the pro-toxic 1,2-unsaturated pyrrolizidine alkaloids. Evidence from the plant-insect interphase is presented indicating a substantial role of the 1,2-saturated alkaloids in plant and insect defense.     

Pyrrolizidine alkaloids from Anchusa strigosa and their antifeedant activity

The pyrrolizidine alkaloid (PA) content of flowers, leaves, and roots of Anchusa strigosa (Boraginaceae) was analysed by ESI-LC-MS. Six PAs, including two new natural compounds, were detected, characterized by NMR spectroscopy, and quantified in each plant organ. The results indicated that the highest total concentration of PAs was in the leaves (23.63 mg/g of dried part), followed by the flowers (19.77 mg/g), and finally by the roots (1.80 mg/g). All PAs isolated were subjected to Spodoptera exigua and Pieris brassicae larvae. Feeding activity by both herbivore species using a bioassay was inhibited up to circa 75% depending on PA and applied concentration.     

不同种类无机盐对灰毡毛忍冬“渝蕾1号”悬浮培养体系中细胞生长和绿原酸含量的影响

为提高灰毡毛忍冬"渝蕾1号"悬浮培养体系中绿原酸的含量,该研究探讨了B_5培养基中不同浓度的无机盐对灰毡毛忍冬"渝蕾1号"悬浮培养细胞生物量及绿原酸含量的影响,通过在悬浮培养体系中添加不同浓度的无机盐,采用重量法测定灰毡毛忍冬"渝蕾1号"悬浮培养细胞的生物量及采用高效液相色谱法测定绿原酸的含量。结果表明:当硝态氮和铵态氮配比与B_5培养基中硝态氮和铵态氮配比一致时,即NO_3~-/NH_4~+摩尔比值为13∶1时,培养体系有利于细胞的生长和绿原酸的积累。当KNO_3浓度为3.5 g·L~(-1)时,细胞生物量达到最大,为19.26 g·L~(-1);当KNO_3在较低浓度(0.5 g·L~(-1)和1.5 g·L~(-1))时,积累较多的绿原酸。NO_3~-的两项研究结果均与对照浓度(2.5g·L~(-1))有一定的差异。另外,对(NH_4)_2SO_4来说,在高于对照浓度0.134 g·L~(-1),即浓度为0.268 g·L~(-1)时,生物量和绿原酸含量都达到了最大。P、Ca、Mg三种矿质元素的研究结果表明,当Na H_2PO_4·2H_2O浓度为0.10 g·L~(-1)、Ca Cl_2的浓度为0.20 g·L~(-1)时,细胞的生长和绿原酸的积累均可达到最大值;而对Mg~(2+)来说,低浓度促进细胞的生长,高浓度促进绿原酸的积累。兼顾细胞生物量和绿原酸含量两个指标,需选择适中的浓度。这些结果均与对照浓度有一定的差异。这说明灰毡毛忍冬"渝蕾1号"悬浮细胞所需无机盐的浓度与B_5培养基无机盐的浓度有一定的差异,选择适宜的浓度可促进其悬浮细胞的生长及次生代谢产物绿原酸的积累。该研究结果为绿原酸的工业化生产打下了基础。     

Quantification of Aconitum alkaloids in aconite roots by a modified RP-HPLC method

The three Aconitum alkaloids, aconitine (1), mesaconitine (2) and hypaconitine (3), are pharmacologically active but also highly toxic. A standardised method is needed for assessing the levels of these alkaloids in aconite roots in order to ensure the safe use of these plant materials as medicinal herbs. By optimising extraction, separation and measurement conditions, a reliable, reproducible and accurate method for the quantitative determination of all three Aconitum alkaloids in unprocessed and processed aconite roots has been developed. This method should be appropriate for use in the quality control of Aconitum products. The three Aconitum alkaloids were separated by a modified HPLC method employing a C18 column gradient eluted with acetonitrile and ammonium bicarbonate buffer. Quantification of Aconitum alkaloids, detected at 240 nm, in different batches of samples showed that the content of 1, 2 and 3 varied significantly. In general, the alkaloid content of unprocessed roots was higher than that of processed roots. These variations were considered to be the result of differences in species, processing methods and places of origin of the samples.     

Comparative analysis of the chemical profile of wild and cultivated populations of Corydalis saxicola by high-performance liquid chromatography

Studies on the simultaneous determination and chemical fingerprinting of alkaloids in Corydalis saxicola Bunting. (Yanhuanglian) were performed for authentication purposes. Ninety samples prepared from different parts of C. saxicola, including whole plants, roots, stems, leaves and flowers, from wild and cultivated populations, were submitted to quantitative determination and fingerprint analysis. Five major alkaloids, namely, tetradehydroscoulerine, dehydroapocavidine, dehydroisoapocavidine, coptisine and dehydrocavidine, were quantitatively analysed by reversed-phase HPLC with acceptable recoveries (>98.2%). Chemical fingerprinting of C. saxicola was established and involved 11 markers. The results indicated that there were no obvious differences between the chemical profiles of wild and of cultivated C. saxicola populations, and that the mean alkaloid contents of the five marker compounds in cultivated populations were significantly higher than those of the wild plants. The highest content of total alkaloids (up to 28.8 mg/g) was found in roots of C. saxicola. The total alkaloids of the leaves were approximately 50% of those of roots, suggesting that the leaves may be employed as an alternative source of alkaloids. Chemical fingerprints and quantitative HPLC analysis will have a positive impact on the conservation and cultivation of this medicinal plant.     

四川乌头属的修订

四川是我国乌头属Aconitum L.植物的重要分布区。本文对该地区的乌头属植物进行了分类修订。通过文献查阅、野外观察和标本室研究,对乌头属的形态性状在居群间和居群内的变异式样进行了比较分析,发现叶分裂程度、上萼片的形状和花梗毛被等性状是较为可靠的分种依据,但是在个别情况,这些性状也会在同一种的同一居群内或不同居群间发生变化。花瓣和种子的特征对于属下划分有重要意义。本文承认四川乌头属植物有46种11变种,另有2种和1变种暂存疑。29种和22变种降为异名;作出新组合2个,其中包括1个改级新组合。紫乌头A. delavayi Franch.和保山乌头A. nagarum Stapf为四川新分布记录。对康定乌头A. tatsienense Finet &; Gagnep.、川鄂乌头A. henryi Pritz.和狭裂乌头A. refractum (Finet &; Gagnep.) Hand.-Mazz.进行了后选模式的标定。初步澄清了东俄洛乌头A. tongolense Ulbr.和狭裂乌头等种类中长期存在的名实混乱。还给出了各个种的形态描述、地理分布以及分亚属、分系和分种检索表。     

Alkaloid production in Vernonia cinerea: Callus, cell suspension and root cultures

Fast-growing callus, cell suspension and root cultures of Vernonia cinerea, a medicinal plant, were analyzed for the presence of alkaloids. Callus and root cultures were established from young leaf explants in Murashige and Skoog (MS) basal media supplemented with combinations of auxins and cytokinins, whereas cell suspension cultures were established from callus cultures. Maximum biomass of callus, cell suspension and root cultures were obtained in the medium supplemented with 1 mg/L alpha-naphthaleneacetic acid (NAA) and 5 mg/L benzylaminopurine (BA), 1.0 mg/L NAA and 0.1 mg/L BA and 1.5 mg/L NAA, respectively. The 5-week-old callus cultures resulted in maximum biomass and alkaloid contents (750 microg/g). Cell suspension growth and alkaloid contents were maximal in 20-day-old cultures and alkaloid contents were 1.15 mg/g. A 0.2-g sample of root tissue regenerated in semi-solid medium upon transfer to liquid MS medium containing 1.5 mg/L NAA regenerated a maximum increase in biomass of 6.3-fold over a period of 5 weeks. The highest root growth and alkaloid contents of 2 mg/g dry weight were obtained in 5-week-old cultures. Maximum alkaloid contents were obtained in root cultures in vitro compared to all others including the alkaloid content of in vivo obtained with aerial parts and roots (800 microg/g and 1.2 mg/g dry weight, respectively) of V. cinerea.