无机盐和蔗糖浓度对白色紫锥菊不定根生长及次生代谢产物积累影响

利用组织培养技术结合高效液相色谱法,考察了液体悬浮培养中无机盐和蔗糖浓度对白色紫锥菊不定根生长以及紫锥菊苷、菊苣酸、氯原酸和酚类化合物积累的影响.结果表明:白色紫锥菊不定根在高或低浓度无机盐和蔗糖培养基中的生长及次生代谢产物含量均较低,不定根生长最适培养基为0.75 MS +5％蔗糖,培养30 d后不定根中紫锥菊苷含量为7.40 mg/g DW,菊苣酸为3.96 mg/g DW,氯原酸含量达3.79 mg/g DW,总酚含量达25.62 mg/g DW.本研究为进一步大规模培养富含紫锥菊苷、菊苣酸的白色紫锥菊不定根奠定了理论和实践基础.     

大孔吸附树脂对紫锥菊提取物中菊苣酸分离纯化的研究

采用大孔吸附树脂分离纯化紫锥菊提取物中免疫活性成分菊苣酸,7％(v／v)的甲醇-水溶液洗脱,HPLC法对产品中菊苣酸含量进行检测分析。该法能把紫锥菊提取物中菊苣酸含量(4％)提高9倍左右,回收率达到95％以上,且操作简单,可用于菊苣酸的分离纯化。     

新疆菊苣籽中总黄酮提取工艺研究

目的:以芦丁为标准品,采用分光光度法测定菊苣籽中总黄酮含量 .方法:在提取过程中通 过单因素实验,分析了乙醇浓度、回流温度、提取时间及料液比等四个因素对提取 率的影响.在单因素实验的基础上建立正交实验,优化菊苣籽总黄酮提取工艺条件.结果: 在最佳吸收波长512nm处,菊苣籽最佳提取工艺条件为乙醇浓度70%,料液比1∶50, 回流温 度70℃,提取时间2h,最佳黄酮含量C为3.80%.结论:以芦丁为对照品 ,用紫外可见分光光度法测定菊苣籽中总黄酮含量,方法简单,准确度较高.     

加热回流法提取淫羊藿苷的试验研究

目的:考察加热回流法提取次数、提取时间、料液比以及乙醇体积分数等实验条件可能对淫羊藿苷的提取率所产生的影响。方法:提取溶剂选择为乙醇,采用加热回流法从淫羊藿中提取淫羊藿苷,采用单因素试验考察淫羊藿苷的提取时间、提取次数、淫羊藿的料液比以及溶剂中乙醇体积分数对淫羊藿苷的提取率所产生的影响,在此基础上进行正交试验的设计,优化提取工艺。结果:结果表明优化的加热回流提取法为料液比1:10(m:V,g/ml),提取2次、提取时间为40min,乙醇体积分数75%。     

菊苣基生叶中总黄酮提取工艺的研究

采用分光光度法,以亚硝酸钠、硝酸铝、氢氧化钠为试剂,芦丁为对照品,乙醇回流提取菊苣基生叶中总黄酮.在提取过程中,通过单因素实验分析了乙醇浓度、回流温度、回流时间及料液比等4个主要因素对提取率的影响.在单因素的基础上,通过正交实验,得到最佳提取工艺为:乙醇浓度为70%,料液比1:40,回流温度80℃,回流时间为2h.实验结果可靠,方法简便,最佳条件适合批量生产中该药材的提取.     

羌活药材活性成分的提取工艺研究

选择提取次数、溶剂倍数、提取时间为考察因素,以提取浸膏量及三种主要活性物质(紫花前胡苷、异欧前胡素、羌活醇)含量为考察指标,探讨羌活药材活性成分的乙醇加热回流提取工艺。在首先考察单因素对提取效果影响的基础上采用正交试验法,优选出羌活药材活性成分的最佳提取工艺参数:加6倍量95%乙醇,加热回流提取3次,每次提取时间为1 h。     

大黄中大黄酸提取工艺研究

以大黄酸含量为参考指标,通过单因素实验分别考察硫酸浓度(A)、液料比(B)、提取加热时间(C)、提取加热温度(D)对大黄中大黄酸提取量的影响,并根据单因素试验结果设计正交试验。结果表明,影响大黄中大黄酸提取工艺的主次因素为:加热温度D>加热时间C>硫酸浓度A>液料比B,大黄中大黄酸的最佳提取工艺为:15%硫酸、5∶1液料比、55℃加热回流2.5 h。此法操作性强,优选出最佳提取工艺条件,为开发大黄中泻下有效部位提供了理论依据。     

纤维素酶-乙醇协同提取黄芩总黄酮

以黄芩总黄酮的得率为指标,比较了乙醇回流提取、酶-缓冲水溶液提取、纤维素酶-乙醇提取等方法对黄芩总黄酮提取的影响,发现纤维素酶对细胞壁的水解和乙醇溶液对黄酮类物质的浸出产生了协同效应。采用单因素实验法,优化了纤维素酶-乙醇协同提取黄芩总黄酮的工艺条件。结果表明,乙醇体积分数50%,提取温度50℃,提取时间6 h,加酶量50U/g(干原料),液固比15时,纤维素酶-乙醇回流法提取黄芩总黄酮的得率达19.85%,较单一的乙醇回流法提取提高了1.38%。纤维素酶-乙醇回流法可用于黄芩总黄酮的提取。     

竹柏叶总黄酮不同提取方法比较

本研究采用正交试验设计分别优选回流提取和超声波提取竹柏叶总黄酮的最佳工艺条件。在单因素试验的基础上,以总黄酮得率为参考指标,选取液料比、乙醇体积分数、温度和时间4个影响因子,进行正交试验,分别确立回流提取和超声波提取最佳提取工艺条件。结果表明,回流提取优化工艺条件为:液料比30∶1(m L∶g),乙醇体积分数为70%,回流时间1.0 h,回流温度80℃;超声波提取优化工艺为:液料比25∶1(m L∶g),乙醇体积分数为70%,超声时间30 min,超声水浴温度60℃。两种提取方法的最终提取效果比较,回流提取的总黄酮得率为15.522%,超声波提取总黄酮得率为13.637%,回流提取法总黄酮得率高于超声波提取法。本实验结果为后续竹柏叶总黄酮的开发利用提供了参考。     

苦参总生物碱提取工艺优化及杀蚜虫活性研究

目的研究苦参总生物碱的提取工艺及杀虫生物活性.方法以总生物碱含量为指标考察乙醇冷浸、乙醇渗漉、乙醇回流三种提取方法的提取效率差异,同时用正交设计对乙醇回流工艺条件进行优化,并以蚕豆蚜虫为供试靶标,探讨苦参总生物碱的杀虫生物活性.结果乙醇回流提取效果较好,液料比、乙醇浓度、提取次数对提取效果均有影响,其最佳提取条件为液料比121,乙醇浓度为60%,提取次数3次;苦参总生物碱对蚕豆蚜虫有一定的杀虫活性,其中氯仿直接提取物对蚜虫杀虫活性最好,随着总生物碱浓度降低,杀虫活性有下降.     

苦丁茶中绿原酸及其异构体的提取变化分析

以富含绿原酸类成分的苦丁茶(Ilex kaushue)为材料,使用溶剂甲醇、乙醇、丙酮和水,结合超声波提取、水浴提取、回流提取等方法对绿原酸及其异构体的提取效率及提取后各异构体的变化进行分析。应用超高效液相色谱法可使苦丁茶的6种绿原酸类成分及咖啡酸在6 min内实现分离。提取结果表明,丙酮作为提取溶剂,在采用超声波和水浴提取时能获得较高的提取效率,易获得较多总量的绿原酸类成分和高含量异绿原酸A。而最常用的溶剂乙醇并未达到理想的提取效果。在不同溶剂的回流提取中,虽然提取的绿原酸类成分总量接近,但异绿原酸A和异绿原酸C含量有较大差异。醇溶液特别是乙醇溶液的回流提取使异绿原酸C的量大幅增加,而相应的异绿原酸A的量大幅减少,表明在醇加热条件下,异绿原酸A转化为异绿原酸C,这为获得抗氧化性更强的异绿原酸C提供了新思路。     

Genetic diversity in seed populations of Echinacea purpurea controls the capacity for regeneration, route of morphogenesis and phytochemical composition

The production of new varieties and higher quality products from Echinacea spp. requires a greater understanding of the regulation of plant growth and the production of specific phytometabolites. The current studies were designed to generate elite varieties of Echinacea purpurea based on regeneration efficiency and chemical profile. Clonal propagation of seedling-derived regenerants and screening for antioxidant potential and concentrations of caftaric acid, chlorogenic acid, cichoric acid, cynarin, and echinacoside identified 58 unique germplasm lines. Chemical profiles varied significantly among germplasm lines but were consistent within clones of each line. In temporary immersion bioreactors, exogenous application of the auxin indolebutyric acid significantly increased the cichoric acid and caftaric acid concentration in the root tissues. Together, these demonstrate the potential for selective breeding of elite, highly regenerative, chemically superior, clonally propagated varieties from the naturally occurring genetic variability in the seed populations of E. purpurea.     

Application of an airlift bioreactor system for the production of adventitious root biomass and caffeic acid derivatives of Echinacea purpurea

In this study, we evaluated the feasibility of using mass cultivation of the adventitious roots of Echinacea purpurea in balloon type bubble (air-lift) bioreactors to produce caffeic acid derivatives, which have pharmaceutical and therapeutic values. An approximately 10 fold increase in biomass and secondary compounds was observed after 4 weeks of culture in balloon type bubble bioreactors (5 L capacity containing 4 L of half strength MS medium). In addition, a linear relationship was observed between the concentration of biomass and the sucrose and ion consumption rate. Furthermore, the concentration of biomass in the bioreactor culture was found to increase as the conductivity decreased. An inoculum density of 7 g/L FW and an aeration rate of 0.1 vvm were found to be suitable for inducing the accumulation of biomass and secondary metabolites. Of the three caffeic acid derivatives evaluated (caftaric acid, chlorogenic acid, and cichoric acid), the concentration of cichoric acid was the highest (26.64 mg/g DW).     

Elicitation of secondary metabolism in Echinacea purpurea L. by gibberellic acid and triazoles

In vitro bioreactor production of Echinacea purpurea L. was used to facilitate the screening of compounds capable of eliciting increased secondary metabolite production. Based on previous experience with various bioreactors, the Southern Sun Liquid Lab Rocker was selected for this study as it produces healthy, vigorous, whole plants. The focus of the present study was to quantify the concentration of the medicinally important secondary metabolites caftaric acid and cichoric acid and to study the effect of growth regulators on the production of these compounds in this system. Both marker compounds were produced at levels that compared favorably to field grown plants. Application of gibberellic acid (GA₃), paclobutrazol, uniconazole and a combined treatment of GA₃ with paclobutrazol to the in vitro plants generally increased the concentration of caftaric acid in the roots with little effect on the concentration in the shoots. The concentration of cichoric acid was higher in the roots of treated plants than in the roots of control plants, but lower in corresponding shoots. The present study highlights the use of in vitro production of whole plants as a model system for studying the regulation of plant secondary metabolism in a controlled environment.     

Variability in the composition of anti-oxidant compounds in Echinacea species by HPLC

A fast and reliable HPLC method for the determination of caffeic acid derivatives (caftaric acid, chlorogenic acid, caffeic acid, cynarin, echinacoside and cichoric acid) in various species of the genus Echinacea has been developed. Extraction of root samples by magnetic stirring with 80% methanol aqueous solution at room temperature allowed the complete recovery of all compounds of interest. Root extracts were analysed on a reversed-phase column with gradient elution and photodiode array detection. Caffeic acid derivatives showed differential qualitative and quantitative distributions in Echinacea species. The total amount of phenolic compounds ranged from 33.95 to 0.32 mg/g. The highest contents of caffeic acid derivatives were found in E. paradoxa var. paradoxa, E. paradoxa var. neglecta and E. purpurea, followed by E. angustifolia var. angustifolia, E. simulata, E. pallida and E. laevigata, whilst E. tennesseensis, E. sanguinea and E. atrorubens had low amounts of phenolic compounds. The radical scavenging activities of methanolic extracts of roots of Echinacea species was evaluated in vitro using the DPPH* radical scavenging method. The EC50 values of the samples ranged from 122 to 1223 microg/mL. The radical scavenging activities of the root extracts were correlated with the content of phenolic compounds, with a correlation coefficient (r2) of 0.923.     

银杏叶多酚的机械力辅助提取及其体外抗氧化活性研究

为优化银杏叶多酚提取工艺,通过单因素试验考察填充率、球磨转速、球磨时间、乙醇浓度、料液比、提取温度、提取时间七个因素对机械力辅助提取银杏叶多酚得率的影响,以银杏叶多酚得率为响应值,采用Box-Benhnken三因素三水平响应面设计优化工艺,同时比较了4种提取方法对银杏叶多酚提取得率和抗氧化活性的差异。结果表明,机械力辅助提取银杏叶多酚的最佳工艺条件为:填充率26%、球磨转速为400rpm、球磨时间为15min。在此条件下,银杏叶多酚的得率为7.33%。机械力辅助乙醇提取银杏叶多酚得率低于碱水提取法,但是抗氧化活性高于碱水法提取的银杏叶多酚;抗氧化活性与乙醇回流法提取的银杏叶多酚相当,但是提取得率高于乙醇回流法。此提取工艺高效可行,具有一定的参考价值。     

正交试验法优选毛泡桐中乌索酸提取工艺条件

采用正交设计L9(34)方法,考察乙醇浓度(A)、超声时间(B)、超声功率(C)、料液比(D)对乌索酸提取率的影响,用高效液相色谱法测定含量,并与常规提取法进行对比,确定了毛泡桐中乌索酸的最佳超声提取工艺条件。所考察的因素对毛泡桐中乌索酸提取的影响按各因素作用主次顺序为:乙醇浓度>料液比>超声时间>超声功率;乌索酸超声提取的最佳条件为:A3B3C2D1,即毛泡桐叶粉末用6倍量体积分数95%乙醇超声提取2 h,超声功率为200 w。与常规的提取方法相比,超声提取具有提取时间短、操作简单、提取率高、无需加热等优点。优选的工艺条件稳定,操作简便,方法可行,可用于毛泡桐中乌索酸的提取。     

Computational fluid dynamics modeling of mass-transfer behavior in a bioreactor for hairy root culture. II. Analysis of ultrasound-intensified process

Recently, cichoric acid production from hairy roots of Echinacea purpurea was significantly improved by ultrasound stimulation in an airlift bioreactor. In this article, the possible mechanism on ultrasound-intensified hairy root culture of E. purpurea in the bioreactor was elucidated with the help of computational fluid dynamics (CFD) simulation, membrane permeability detection, dissolved oxygen concentration detection, confocal laser-scanning microscopy (LSM) observation, and phenylalanine ammonium lyase (PAL) activity analysis. The CFD model developed in Part I was used to simulate the hydrodynamics and oxygen mass transfer in hairy root bioreactor culture stimulated by ultrasound. A dynamic mesh model combined with a changing Schmidt number method was used for the simulation of the ultrasound field. Simulation results and experimental data illustrated that ultrasound intensified oxygen mass transfer in the hairy root clump, which subsequently stimulated root growth and cichoric acid biosynthesis. Ultrasound increased the hairy root membrane permeability, and a high root membrane permeability of 0.359 h(-1) was observed at the bottom region in the bioreactor. LSM observation showed that the change in the membrane permeability recovered to normal in the further culture after ultrasound stimulation. PAL activity in the hairy roots was stimulated by ultrasound increase and was correlated well to cichoric acid accumulation in the hairy roots of E. purpurea.     

同时提取光果甘草中光甘草定和甘草酸的工艺研究

本文主要对光果甘草中光甘草定、甘草酸两种重要活性成分的同时提取工艺进行了研究。通过单因素试验确定了影响提取的主要因素及适宜水平范围,通过正交试验确定了其最佳提取条件为:用含0.6%氨水的60%乙醇溶液作为提取剂,料液比为1∶20(w:v),在75℃条件下提取2 h。最佳提取条件下光甘草定的得率为0.238%±0.002%,甘草酸的得率为5.08%±0.03%。该工艺操作简单,投入一批原料,即可同时提取其中的光甘草定和甘草酸,为实现甘草资源高附加值的综合加工利用奠定了基础。