大孔吸附树脂纯化山蜡梅叶中总黄酮的研究

目的 研究大孔树脂纯化山蜡梅叶中总黄酮的工艺条件及参数.方法 采用静态吸附-解吸方法,以吸附量和解吸率为指标,筛选最佳树脂;又以总黄酮质量浓度为指标,考察了最佳树脂纯化山蜡梅叶中总黄酮的工艺参数.结果 8种树脂中,HPD400树脂对山蜡梅叶中总黄酮纯化效果较好,其总黄酮的静态吸附量达到17.77 mg/g树脂,解吸率为92.24%;动态吸附量为1.68 g/g树脂,用4倍柱体积蒸馏水、4倍柱体积30%乙醇洗脱除去杂质后,换用70%乙醇6倍柱体积洗脱,总黄酮质量分数为31.4%,总黄酮转移率为88.4%.结论 HPD400型大孔树脂在所确定的工艺条件下,可较好地纯化山蜡梅叶总黄酮.     

大孔吸附树脂纯化大米草总黄酮的研究

目的 研究大孔树脂纯化大米草总黄酮的工艺条件及参数.方法 采用静态吸附-解吸方法,以吸附量和解吸率为指标,筛选最佳树脂;又以总黄酮质量浓度为指标,考察了最佳树脂纯化大米草总黄酮的工艺参数.结果 5种树脂中,Hz-816树脂对大米草总黄酮纯化效果较好,其总黄酮的静态吸附量达26.79mg/g,解吸率为90.10%;用4倍柱体积蒸馏水、2倍柱体积10%乙醇洗脱除去杂质后,换用50%乙醇4倍柱体积洗脱得纯化后的大米草总黄酮溶液,总黄酮质量分数为25.36%.结论 Hz-816型大孔树脂在所确定的工艺条件下,可较好地纯化大米草总黄酮.     

失效模式分析和星点设计-效应面法优化熟三七皂苷类成分纯化工艺

目的优化熟三七中皂苷类成分的大孔吸附树脂纯化工艺。方法通过失效模式分析(FMEA)筛选纯化工艺的主要影响因素,采用星点设计-效应面法(CCD-RSM),以皂苷类成分的回收率和质量分数为评价指标,对大孔吸附树脂纯化工艺进行优选。实验中以5个主要影响因素(上柱液质量浓度、上样体积、水洗体积、乙醇体积分数、乙醇洗脱体积)考察了熟三七皂苷类成分的纯化工艺。结果以大孔吸附树脂为纯化工艺,优化工艺为当上柱液质量浓度为11.22 mg/mL,上样体积为4.97 BV,水洗体积为2 BV,乙醇体积分数为70%,乙醇洗脱体积为3.31 BV时,可获得最大的皂苷回收率(82.81%)和皂苷质量分数(77.24%)。结论优化的纯化工艺简单、稳定,熟三七皂苷类成分回收率和质量分数均较高,具有一定的实用价值。     

大孔树脂纯化黄蜀葵花总黄酮的工艺优选

目的:优选黄蜀葵花总黄酮的大孔树脂纯化工艺,为黄蜀葵花制剂的工业化生产提供参考。方法:利用静态吸附-洗脱试验筛选纯化黄蜀葵花总黄酮的大孔树脂型号。采用UV测定总黄酮含量,检测波长510 nm;在单因素试验基础上,以总黄酮质量及纯度的综合评分为指标,采用正交试验考察乙醇体积分数、上样液质量浓度及p H对纯化工艺的影响。结果:AB-8型大孔树脂对黄蜀葵花总黄酮的吸附与洗脱性能较好。最优纯化条件为树脂柱径高比1∶6,上样液质量浓度0.2 g·m L-1,p H 5.0,最佳上样量3 BV,乙醇体积分数75%,洗脱液用量6 BV;总黄酮质量417.5 mg,纯度58.04%。结论:AB-8型大孔树脂可有效富集、纯化黄蜀葵花总黄酮,优选的纯化工艺稳定可行。     

卷柏总黄酮纯化工艺研究

目的:研究聚酰胺富集卷柏总黄酮的工艺参数.方法:以总黄酮和阿曼托双黄酮的含量以及总黄酮的收率为考察指标,经聚酰胺吸附柱富集后,分别采用紫外分光光度法和高效液相色谱法测定.结果:纯化工艺条件为:卷柏提取物与聚酰胺比为50:3(mg/mL),树脂径高比为1:6,用10倍量柱体积的50%乙醇除杂,用6倍量柱体积的80%乙醇洗脱,洗脱流速为1.0 mL/min.结论:本法富集纯化卷柏总黄酮工艺稳定可行.     

大孔树脂制备岩柏草总黄酮及其含量测定

目的:探索D-101大孔树脂分离纯化岩柏草总黄酮的最优工艺,并对有效成分穗花杉双黄酮进行含量测定。方法:紫外分光光度法测定总黄酮浓度,以黄酮收率、纯度等为指标,上样质量浓度、吸附流速和洗脱剂乙醇体积分数为影响因素,确定大孔树脂分离纯化岩柏草总黄酮工艺,并用高效液相色谱法测定总黄酮中穗花杉双黄酮。结果:大孔树脂对岩柏草总黄酮有较好的分离效果,其最优工艺条件为上样质量浓度8.5mg/mL,吸附流速为2mL/min,乙醇体积分数为70%,总黄酮得率为0.89%。流动相:乙腈-0.5%冰醋酸溶液,梯度洗脱;检测波长:338nm;流速:1mL/min,用HPLC法测得穗花杉双黄酮含量为56%。结论:用D-101大孔树脂分离纯化岩柏草总黄酮简单可行,精制效果好。用上述色谱条件可用于穗花杉双黄酮的质量控制。     

千里光总黄酮的分离纯化研究

目的 研究大孔吸附树脂分离纯化千里光总黄酮的工艺.方法 以千里光总黄酮质量浓度、回收率等为考察指标,选用大孔吸附树脂对千里光总黄酮进行分离纯化.分别采用静态试验、动态试验考察大孔树脂对千里光总黄酮的分离纯化效果及影响因素.结果 AB-8型大孔吸附树脂为最佳树脂,洗脱剂为70%乙醇.洗脱剂用量为4倍树脂体积,体积流量3～4 mL/min;上柱总黄酮质量与树脂质量比为1:9.4,上柱液总黄酮质量浓度为17.86 mg/mL,体积流量2～3 mL/min;冲洗杂质用水体积2～3 BV,体积流量2～3 mL/min;上柱液pH 4～5;径高比1.5:21.6.AB-8型大孔吸附树脂对千里光总黄酮静态饱和吸附量为111.25 mg/g,洗脱率91.53%,动态饱和吸附量为96.8mg/g,洗脱率为93.40%,总黄酮回收率、质量分数均在90%以上.结论 采用大孔吸附树脂对千里光中的总黄酮进行纯化可行,可以为合理的开发利用千里光提供参考.     

山黄降脂片总黄酮富集工艺研究

目的:对山黄降脂片降脂有效部位的富集工艺进行研究。方法:采用亚硝酸钠-硝酸铝-氢氧化钠比色法测定总黄酮含量、HPLC测定葛根素含量,以正交试验法和单因素试验法筛选提取工艺和大孔吸附树脂分离条件。结果:最佳提取工艺为采用12倍量60%乙醇回流提取,提取温度为90℃,提取时间为120min。纯化工艺为将提取物上D-101型大孔吸附树脂,用5倍柱体积的水洗脱,再用12倍柱体积的70%乙醇以3mL/min流速进行洗脱,收集洗脱液。结论:D-101型大孔吸附树脂对山黄降脂片中总黄酮的富集能力优于其他树脂。洗脱溶剂70%乙醇,洗脱12倍柱体积能较好地洗脱黄酮类成分。     

大孔树脂纯化复方四叶减肥汤中总黄酮成分的研究

目的:初步研究大孔树脂纯化复方四叶减肥汤中总黄酮成分的工艺条件和参数。方法:以静态吸附试验、动态吸附试验、比吸附量、比洗脱量等为研究指标,比较不同大孔树脂纯化复方中总黄酮的性能,对树脂纯化工艺进行正交优选。结果:AB-8型树脂为优选树脂,最佳工艺条件为:树脂(g)-提取液(mL)比例为1.2∶1上柱,色谱柱径高比1∶12,水洗至无色,80%乙醇洗脱液收集量为7 BV。结论:该工艺稳定性良好,纯化后的总黄酮质量分数达85%以上。     

基于灰色关联度分析的荆芥抗Caco-2细胞总黄酮纯化工艺研究

目的通过灰色关联度分析技术及对Caco-2肠癌细胞增殖情况的影响来评价荆芥总黄酮大孔吸附树脂纯化工艺。方法以总黄酮静态吸附量和解吸率为考察指标筛选大孔吸附树脂型号,采用单因素试验,优化其纯化总黄酮的工艺条件,并采用MTT法测定Caco-2细胞抑制率,同时应用灰色关联度分析技术,计算总黄酮质量分数与对肿瘤细胞增殖抑制率的关联度。结果 HPD-400型大孔吸附树脂对荆芥总黄酮有较好的吸附分离性能,荆芥药材以0.1 g/m L的生药质量浓度、0.8 g/m L(药材/湿树脂)的上柱量上柱,用2倍柱体积(BV)水洗除杂,用8 BV、70%乙醇进行洗脱,体积流量均为4 BV/h,收集洗脱液,挥干,同法进行第2次纯化,总黄酮质量分数为92.15%,对Caco-2细胞增殖抑制率为86.17%,总黄酮质量分数与抑制率灰色关联系数为0.950 2。结论此工艺条件稳定可行,体外抗Caco-2肿瘤细胞药效明确,为开发以荆芥总黄酮纯化物为主成分的抗癌新药奠定基础。     

丛枝菌根真菌与哈茨木霉菌合用对连作丹参生长及质量的影响

该文探讨田间条件下丛枝菌根真菌(AMF)与哈茨木霉菌合用对丹参生长及质量的影响。采用田间小区试验来进行研究,使用HPLC测定丹参地上部分和地下部分有效成分含量,通过形态观察并计算根部发病率,并结合方差统计学方法对各指标的测定结果进行分析。结果显示,单独接种AMF以及AMF与哈茨木霉菌合用能够有效的降低连作丹参根部病害的发生率,其中AMF与哈茨木霉菌合用效果更佳,比CK组降低了61.50%。几种处理对丹参生物量影响没有显著差异,但都能提高丹参根部各种有效成分的含量。单独接种AMF以及AMF+哈茨木霉都能显著提高丹参根中丹酚酸B的含量(P<0.05),其中AMF对丹酚酸B促进作用最明显,提高了27.97%;接种AMF能显著提高丹参根丹参酮I和丹参酮ⅡA的含量(P<0.05),其中丹参酮I约提高了70.14%;丹参酮ⅡA约提高了51.42%。另外,单独接种AMF以及AMF+哈茨木霉均能显著提高丹参根中隐丹参酮的含量,提高了约30.67%。从而得出单独接种AMF以及AMF+哈茨木霉能够有效减低连作丹参病害的发生率,同时提高连作丹参药材的质量。     

列当属、肉苁蓉属和草苁蓉属植物传统药物学调查

通过野外资源调查、整理各大标本馆标本原始记录和查阅文献记载等方法,系统整理、总结、归纳了列当属、肉苁蓉属和草苁蓉属民族药用植物种类、功效及民间使用情况,结果表明列当属6种药用植物在4个少数民族间作为7种民族药应用,草苁蓉属2种药用植物在8个少数民族间作为10种民族药应用,肉苁蓉属2种药用植物在3个少数民族间作为3种民族药应用,且同种药用植物常作不同民族药;发现3属植物的传统疗效主要集中在补肾壮阳、止血和止痛3个方面,并且该传统疗效与现代药理研究结果基本吻合。因此深入研究植物种类丰富的列当属植物资源对缓解肉苁蓉植物资源匮乏局面和扩大药源具有积极意义。     

金银花类药用植物IspE和IspH基因克隆和生物信息学分析

目的:克隆金银花类药用植物4-二磷酸胞苷-2-C-甲基赤藓糖激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,IspE)和4-羟基-3-甲基-2-邻苯基二磷酸还原酶(4-hydroxy-3-methylbut-2-enyl diphosphate reductase,IspH)基因,并对其基因序列、蛋白特性和转录活性进行分析、比较.方法:从忍冬Lonicera japonica转录组测序结果中分析获得了IspE,IspH基因.分别以忍冬、红白忍冬L.japonica var.chinensis、红腺忍冬L.hypoglauca和水忍冬L.dasystyla新鲜花蕾为材料,利用RT-PCR技术克隆获得了4种金银花类药用植物IspE和IspH基因的全长cDNA.运用生物信息学分析软件,预测编码蛋白的结构和功能,并通过RT-PCR检测IspE和IspH在忍冬、红白忍冬、红腺忍冬、水忍冬花蕾中的转录情况.结果:金银花类药用植物IspE基因含有1个完整的开放阅读框,长度为1 221 bp,编码406个氨基酸;IspH含有一个完整的开放阅读框,长度为1 380 bp,编码459个氨基酸.IspE和IspH均为非分泌蛋白,均定位于叶绿体中.RT-PCR分析结果表明在忍冬、红腺忍冬和水忍冬的花蕾中IspE,IspH基因的转录水平没有显著差异,但红白忍冬花蕾中IspE,IspH基因的转录水平均显著高于忍冬.结论:克隆获得忍冬、红白忍冬、红腺忍冬和水忍冬中IspE,IspH基因,并证实了其在不同金银花类药用植物中的表达,为进一步研究IspE,IspH基因对萜类化合物生物合成和花香气以及颜色的影响奠定了基础.     

灰毡毛忍冬褐斑病发生规律与防治方法研究

目的:通过对灰毡毛忍冬褐斑病的发生规律和药剂防治研究,为综合防治提供依据.方法:田间调查和药剂防治试验.结果:褐斑病由Cercospora sp.引起,4月上中旬开始发病、7-8月为发病盛期;发病情况与海拔、土壤有机质含量、土壤速效磷含量、土壤速效钾含量和栽培品种相关;10％苯醚甲环唑水分散粒剂1 500倍液防治效果最好,50％福美双可湿性粉剂800倍液、70％代森锰锌可湿性粉剂800倍液也具有较好防治效果.结论:适当的农业防治和化学防治方法能有效防治灰毡毛忍冬褐斑病.     

髯丝蛛毛苣苔和吊石苣苔抗菌活性成分研究

目的 研究髯丝蛛毛苣苔和吊石苣苔的抗菌活性成分.方法 以小檗碱为阳性对照,利用纸片扩散法进行抗菌活性研究,液体培养法进行活性成分筛选;采用各种色谱法对高活性部位分离,运用多种波谱技术鉴定结构.结果 从髯丝蛛毛苣苔醋酸乙酯部位分离得到5个化合物β-谷甾醇(1),E-3,4-二甲氧基肉桂酸(2),barbinervic acid (3),3β,19α-二羟基-12-烯-28-乌苏酸(4),28-O-β-D-glucopyranosyl pomolic acid(5).从吊石苣苔醋酸乙酯部位分离得到3个化合物 5,7-二羟基-6,8,4'-三甲氧基黄酮(6),5,6,4'-三羟基-7,8-二甲氧基黄酮(7),5-羟基-6,8,4'-三甲氧基黄酮-7-O-β-D-葡萄糖苷(8).化合物3,4,6对金黄色葡萄球菌(SA)、耐甲氧西林葡萄球菌(MRSA)和β-内酰胺酶阳性的金黄色葡萄球菌(ESBLs-SA)具有抑制活性.其中化合物3(IC50 0.098 g·L-1)抑制SA活性最好;化合物4(IC50 0.130 g·L-1)抑制MRSA活性最好;化合物3(IC50 0.270mg·L-1)抑制ESBLs-SA活性最好.结论 化合物1～5为首次从植物中分离得到,7,8为首次从该科植物中分离得到.     

Anthelmintic activity of Arisaema franchetianum and Arisaema lobatum essential oils against Haemonchus contortus

Ethnopharmacological relevance

Arisaema franchetianum and Arisaema lobatum are two perennial plants native to China. Arisaema franchetianum is universally used to promote the subsidence of induration and swelling, quicken blood and relieve pains, and kill intestinal parasites in humans and animals. Arisaema lobatum is used to treat malaria, intestinal parasites, and snake and insect bites in humans and animals. The aim of this study was to determine the composition of the essential oils from Arisaema franchetianum and Arisaema lobatum and evaluate the anthelmintic effect against Haemonchus contortus.

Materials and methods

Two oils were investigated by GC and GC–MS. The anthelmintic bioassay tests of Arisaema franchetianum and Arisaema lobatum essential oil, linalool and carvacrol were performed using egg hatch assay (EHA), larval development assay (LDA) and larval migration inhibition assay (LMIA).

Results

Fifty six components representing 96.88% of the Arisaema franchetianum oil and 64 components representing 96.88% of the Arisaema lobatum oil were identified. Carvacrol and linalool were found to be the major constituents of two oils. In the EHA, greater than 99% inhibition were observed with Arisaema franchetianum oil at 10 mg/mL (CE₅₀ 1.63 mg/mL) and Arisaema lobatum oil at 5 and 10 mg/mL (CE₅₀ 0.48 mg/mL). In the LDA, both oils induced complete inhibition at 10 mg/mL, with the CE₅₀ being 1.10 mg/mL for Arisaema franchetianum oil and 0.73 mg/mL for Arisaema lobatum oil. In the LMIA, the Arisaema franchetianum oil and Arisaema lobatum oil at best inhibited 74.1% and 95.6% of larval migration at 10 mg/mL, respectively. Carvacrol exhibited similar activity to Arisaema lobatum essential oil and linalool did not show high activity in every assay.

Conclusions

These data show for the first time that the essential oils obtained from Arisaema franchetianum or Arisaema lobatum had promising anthelmintic activity against Haemonchus contortus. Arisaema plant may offer an alternative source for the control of gastrointestinal nematodes of sheep and goats.     

不同寄主锁阳种子质量差异及相互寄生关系研究

以自然条件下充分成熟的唐古特白刺、泡果白刺、西伯利亚白刺、霸王、骆驼蓬上寄生的锁阳种子为研究对象,研究其种子形态特征、胚率、生活力、千粒重、净度、含水量的差异及不同寄主种子的相互寄生关系。结果表明,不同寄主锁阳种子在形态、颜色及表面状态极其相似,根据种子形态,无法对其寄主进行判断,对于寄主为唐古特白刺、西伯利亚白刺、骆驼蓬上的锁阳种子,可选用稍小于0.923 1 mm和稍大于1.066 2 mm的圆孔筛清选种子,对于寄生于泡果白刺上的锁阳种子,可选用稍小于0.926 1 mm和稍大于0.985 3 mm的圆孔筛清选种子,对于寄生于霸王上的锁阳种子,可选用稍小于0.751 3 mm和稍大于1.035 3 mm的圆孔筛清选种子;锁阳种子形态指标之间极显著相关(P<0.01),形态指标与千粒重之间显著相关(0.01<P<0.05),而与生活力和胚率相关性不显著,千粒重与生活力和胚率不相关,但生活力和胚率极显著相关(P<0.01);完全成熟的各寄主的锁阳种子生活力和优良度很高,含水量很低,净度较低,在质量评定和分级时应考虑生境和寄主间的差异;锁阳种子对寄主无选择性,不同寄主的锁阳种子可以在其他寄主上寄生。     

Antimalarial and safety evaluation of Pluchea lanceolata (DC.) Oliv. & Hiern: In-vitro and in-vivo study

Ethnopharmacological relevance

Many of the effective therapeutic strategies have been derived from ethnopharmacologically used natural products. Pluchea lanceolata is an herb employed in Indian folk medicine for malaria like fever but it lacks proper pharmacological intervention.

Aim of the study

To evaluate antimalarial and safety profile of Pluchea lanceolata: an in-vitro, in-vivo for its ethnopharmacological validation.

Materials and methods

Methanol, butanol, ethyl acetate, chloroform, hexane extracts and its isolate, taraxasterol acetate (TxAc) were obtained from air dried aerial part of Pluchea lanceolata. These were tested in-vitro against chloroquine-sensitive strain of Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. The most potent hexane extract and TxAc were further validated for in-vivo antimalarial and safety evaluation. TxAc, a pentacyclic-triterpene isolated from the most active fraction was further evaluated with special emphasis on inflammatory mediators involved in malaria pathogenesis. Murine malaria was induced by intra-peritoneal injection of Plasmodium berghei infected red blood cells to the male Swiss inbred mice. Mice were orally treated following Peters 4-Day suppression test. In-vivo antimalarial efficacy was examined by evaluating the parasitaemia, percent survival, mean survival time, blood glucose, haemoglobin and pro-inflammatory mediators involved in malaria pathogenesis.

Results

Hexane extract and TxAc showed promising antimalarial activity in-vitro and in-vivo condition. TxAc attributed in inhibition of the pro-inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle treated infected mice. We have not observed the synergistic action of combinations of chloroquine and TxAc from our experimental results. In-vitro and in-vivo safety evaluation study revealed that hexane extract is non toxic at higher concentration.

Conclusion

Present study further validates the ancient Indian traditional knowledge and use of Pluchea lanceolata as an antimalarial agent. Study confirms the suitability of Pluchea lanceolata as a candidate for further studies to obtain a prototype for antimalarial medicine.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

光照强度对北细辛生长发育及质量的影响

目的:对给予100%(Ⅰ),50%(Ⅱ),28%(Ⅲ),12%(Ⅳ)自然光照条件的五年生北细辛的展叶期,花期,果初期,果熟期,枯萎期,叶斑病发生程度及发生率,根折干率,挥发油含量及组分进行调查,分析和测试.方法:依照<中国药典>(2010年版)一部规定的挥发油提取方法-XD法对不同光照条件下北细辛根的挥发油进行提取,采用气相-质谱联用仪对不同光照条件下根挥发油组分进行分析.结果:不同光照条件,北细辛的展叶期,花期,结果初期等均随光照强度减弱而推迟2 d左右;Ⅰ～Ⅳ4种光照条件下的叶斑病发病率分别为88.46%,70.00%,0.23%,0.07%,折干率分别为25.14%,28.27%,30.23%,31.57%,挥发油分别为18.1,17.6,16.3,15.3 mL·kg-1,4个光照条件下的挥发油总离子流图存在差异;挥发油的组分类别及含量均存在差异,各组分的含量并未出现随光照强度的减弱呈现逐渐减少/增加的趋势,其中光照条件Ⅲ下,甲基丁香酚最高为46.75%,黄樟醚最低为11.42%.结论:光照条件对北细辛的展叶期,花期,结果初期均有影响,北细辛叶斑病发生率和发生程度均随光照强度减弱而减轻,根折干率随光照强度减弱而增大,挥发油含量随光照强度减弱而减小,同时,光照强度对挥发油的组分类别及含量亦有影响.