HPLC测定灯盏花不同部位绿原酸、灯盏乙素、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸含量

目的: 建立同时测定灯盏花不同部位中绿原酸、灯盏乙素、3,5-二咖啡酰奎宁酸及4,5-二咖啡酰奎宁酸4种有效成分含量的高效液相色谱方法。 方法: 采用Agilent Zorbax SB-C₁₈ (4.6 mm×250 mm, 5 μm) 色谱柱,流动相为乙腈(A)-0.3% 磷酸水溶液(B),梯度洗脱(0~10 min,12%~15% A,10~32 min,15% ~15% A,32~33 min,15%~20% A,33~ 50 min,20%~22% A),流速1 mL·min^-1,检测波长335 nm与327 nm,柱温30℃。 结果: 绿原酸(1)、灯盏乙素(2)、3,5-二咖啡酰奎宁酸(3)及4,5-二咖啡酰奎宁酸(4)分别在0.050 1~1.002,0.165 9~3.318,0.049 7~ 0.994,0.048 7~0.974 μg呈良好线性关系(r>0.999 6);灯盏花药材中4种有效成分的平均回收率(n=6)分别为98.53%,99.68%,98.78%,99.06%,RSD 分别为0.94%,0.49%,1.1%,0.81%。 结论: 该方法简便、准确,分离效果好,可用于灯盏花药材的质量评价。     

基于指纹图谱、模式识别及多成分同时定量的野菊花质量评价研究

目的建立HPLC指纹图谱、模式识别、多成分同时定量的质量评价模式及其在野菊花药材质量评价中的可行性及有效性分析。方法建立21批次野菊花HPLC指纹图谱,通过TCMYS指纹图谱软件对不同产地野菊花指纹图谱进行相似度分析、主成分分析及聚类分析,对指纹图谱中12个共有色谱峰进行指认,采用外标法对12种成分进行定量。结果 21批次野菊花HPLC指纹图谱相似度较高,有分类趋势,标定了17个共有峰,其中12个共有峰被指认为5-咖啡酰奎宁酸、绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、木犀草苷、4,5-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、蒙花苷、木犀草素和芹菜素和金合欢素,外标法线性关系良好(r≥0. 999 6),加样回收率均在95. 84%~102. 46%之间,RSD <2. 72%。结论 HPLC指纹图谱、模式识别、多成分同时定量的方法准确、可靠,为野菊花全面质量评价提供重要参考依据。     

昆仑雪菊与杭菊、贡菊主要活性成分比较

用RP-HPLC法测定昆仑雪菊和杭菊、贡菊中绿原酸、3,5-O-二咖啡酰奎宁酸、木犀草苷和槲皮苷的含量。结果表明,昆仑雪菊和杭菊、贡菊主要活性成分相似,但不同活性成分含量差异显著,其中昆仑雪菊中绿原酸、木犀草苷及槲皮苷含量最高,质量分数分别为7.46,46.58,26.01 mg·g^-1,绿原酸和木犀草苷分别高于《中国药典》(2010年版)规定的2.00,0.80 mg·g^-1。杭菊中3,5-O-二咖啡酰奎宁酸含量最高,质量分数为14.70 mg·g^-1。通过该实验对昆仑雪菊活性的成分测定,为昆仑雪菊进一步研究、开发提供依据。     

HPLC法测定杏香兔耳风中绿原酸, 5-二咖啡酰奎宁酸和木犀草素

目的建立杏香兔耳风中绿原酸、3,5-二咖啡酰奎宁酸和木犀草素的测定方法。方法采用高效液相色谱法测定杏香兔耳风中绿原酸、3,5-二咖啡酰奎宁酸和木犀草素的量。色谱条件为DiamonsilTMC18柱(250 mm×40 mm,5μm);流动相甲醇(A)-0.1%磷酸水(B),梯度洗脱(0～15 min,28%A;15～25 min,28%～40%A;25～50 min,40%A);体积流量1.0 mL/min;检测波长328 nm;柱温室温。结果杏香兔耳风中绿原酸、3,5-二咖啡酰奎宁酸和木犀草素的线性范围分别为0.132～1.32、0.078～0.78、0.034～0.34μg,平均加样回收率分别为99.1%、103.1%、99.3%。结论该方法快速简便、精密度好、灵敏度高,可用于杏香兔耳风中绿原酸、3,5-二咖啡酰奎宁酸和木犀草素的同时测定。     

一测多评法同时测定野菊花中7种成分

目的建立野菊花Chrysanthemum indicum中绿原酸、咖啡酸、木犀草苷、3,5-O-二咖啡酰基奎宁酸、蒙花苷、木犀草素、芹菜素7种成分一测多评测定方法。方法采用HPLC法,以绿原酸为内参物,建立绿原酸与咖啡酸、木犀草苷、3,5-O-二咖啡酰基奎宁酸、蒙花苷、木犀草素、芹菜素的相对校正因子,并考察了相对校正因子的重现性,分别采用外标法和一测多评法进行野菊花中7种成分的测定。结果建立的校正因子重现性良好,野菊花药材采用一测多评法计算的量值与外标法实测值之间没有明显差异。结论同时测定7种成分的一测多评法控制野菊花药材的质量是可行、准确的。     

野菊花心血管活性部位化学成分的研究

目的:研究菊科植物野菊花心血管活性部位中的主要化学成分.方法:采用多种色谱方法进行分离纯化,通过波谱方法结合文献对照鉴定化合物结构.结果:从野菊花心血管活性部位中分离并鉴定了12个化合物,包括7个黄酮类化合物(1～7),5个苯丙素类化合物(8～12),分别鉴定为(2S)-圣草酚-7-O-β-D-吡喃葡萄糖醛酸苷(1),(2S)-圣草酚-7-O-β-D-吡喃葡萄糖苷(2),(2S)-橙皮素-7-O-β-D-吡喃葡萄糖醛酸苷(3),木犀草素-7-O-β-D-吡喃葡萄糖苷(4),木犀草素-7-O-β-D-吡喃葡萄糖醛酸苷(5),香叶木素-7-O-β-D-吡喃葡萄糖醛酸苷(6),槲皮素-7-O-β-D-呲喃葡萄糖苷(7),3,5-二咖啡酰奎宁酸甲酯(8),3,5-二咖啡酰奎宁酸(9),3,5-二顺式咖啡酰奎宁酸(10),1,5-二咖啡酰奎宁酸(11),1,3-二咖啡酰奎宁酸(12).结论:野菊花心血管活性部位中主要成分为黄酮类化合物,共5个化合物分别为2～3,7～8,10;共10个化合物分别为2～3,5～12系首次从该种植物分离得到.     

一测多评法测定山银花提取物中6种绿原酸类成分

以绿原酸为内标,测定新绿原酸、隐绿原酸、3,4-O-双咖啡酰奎宁酸、3,5-O-双咖啡酰奎宁酸和4,5-O-双咖啡酰奎宁酸的相对校正因子,建立山银花提取物中6种绿原酸类成分的一测多评定量分析方法。通过考察不同色谱仪和不同色谱柱以验证相对校正因子的耐用性,同时采用外标法对4批山银花药材及20批山银花提取物进行分析,并将其含量测定结果与一测多评法测定结果进行比较。结果表明相对校正因子的耐用性良好,4批山银花药材及20批山银花提取物一测多评法与外标法所得结果无显著差异。所建立的含量测定方法可用于同时测定山银花提取物中6种绿原酸类成分。     

UPLC-MS同时测定蒙药材蓝刺头中6个成分的含量

目的:建立超高效液相色谱-串联质谱法(UPLC-MS)同时测定蒙药材蓝刺头中绿原酸、1,5-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、芹菜素和芹菜素-7-O-葡萄糖苷含量的方法。方法:样品分析采用Agilent 6430液质联用仪;色谱柱为Agilent ZORBAX RRHD Ecilipse Plus-C_(18);流动相为0.1%甲酸水溶液-甲醇,梯度洗脱;离子源为电喷雾电离源(ESI);采用负离子多反应监测模式(MRM)。结果:绿原酸、3,5-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-葡萄糖苷、4,5-二咖啡酰奎宁酸和芹菜素分别在1.25~40.00(r=0.999 0)、0.94~30.00(r=0.999 8)、0.94~30.00(r=0.999 8)、0.16~5.00(r=0.999 7)、0.16~5.00(r=0.999 9)、0.02~2.50(r=0.999 5)μg·mL~(-1)线性关系良好;加样回收率为99.80%~102.97%,RSD为1.23%~2.98%。结论:该方法专属性强、精密度高、重复性好,为蒙药材蓝刺头的成分研究提供科学参考。     

中药黄金菊汤剂君药的6种抗炎药效组分含量测定

目的：分析中药黄金菊君药野菊花中6种抗炎组分含量。方法：反相高效液相色谱法（RP—HPLC）。结果：分析得到了君药中6种抗炎组分含量,即绿原酸,术犀草素．木犀草苷．蒙花苷,3,5二咖啡酰奎宁酸。结论：野菊花在黄金菊汤剂中的抗炎药效组分为绿原酸-咖啡酸-木犀草素-木犀草苷-蒙化苷-3．5二咖啡酰奎宁酸（10：1：1：4：17：6）．为组分中药的发现和中药药效组分质量评价方法的建立提供了科学基础数据。     

金银花中抗补体活性酚酸类成分的研究

目的: 研究金银花Lonicera japonica中的抗补体活性酚酸类成分. 方法: 通过溶血试验方法进行抗补体活性成分的导向分离,对所得化合物进行抗补体活性测定,采用现代波谱技术ESI-MS,¹H-NMR,¹³C-NMR进行结构鉴定. 结果: 从金银花乙酸乙酯部位分离得到了14个化合物,其中有8个酚酸类、3个环烯醚萜类和3个黄酮类成分,包括新绿原酸(1)、绿原酸(2)、隐绿原酸 (3)、3,5-O-二咖啡酰奎宁酸(4)、4,5-O-二咖啡酰奎宁酸(5)、3,4-O-二咖啡酰奎宁酸(6)、咖啡酸(7)、咖啡酸甲酯(8)、裂环马钱苷(9)、獐芽菜苷(10)、断氧化马钱子苷(11)、木犀草素(12)、槲皮素(13)、山柰酚(14).抗补体实验表明化合物 1~9, 11~14 对经典的补体激活具有不同程度的抑制作用,以3,5-O-二咖啡酰奎宁酸活性最强. 结论:化合物 14 首次从金银花中分离得到,酚酸类化合物是金银花中的主要抗补体活性成分,3,5-O-二咖啡酰奎宁酸活性最强,值得深入研究.     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

真菌的中药干预研究近5年进展

近年来真菌感染率逐年上升,传统抗真菌药物易产生耐药性,而中药在防治真菌感染方面具有一定的优势。本文就近5年来中药对白念珠菌、皮肤癣菌、曲霉菌、马拉色菌、串珠镰孢菌、申克孢子丝菌、新生隐球菌及真菌生物膜的干预研究进展进行综述。     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

α-Glucosidase inhibitory activity of selected Philippine plants

Ethnopharmacological relevance

Antidesma bunius Spreng. (Phyllantaceae), Averrhoa bilimbi L. (Oxalidaceae), Biophytum sensitivum (L.) DC. (Oxalidaceae), Ceriops tagal (Perr.) C.B. Rob. (Rhizophoraceae), Kyllinga monocephala Rottb. (Cyperaceae), and Rhizophora mucronata Lam. (Rhizophoraceae) are used as remedies to control diabetes. In the present study, these plants were screened for their potential α-glucosidase inhibitory activity.

Materials and methods

The 80% aqueous ethanolic extracts were screened for their α-glucosidase enzyme inhibitory activity using yeast alpha glucosidase enzyme.

Results

Except for A. bilimbi with IC₅₀ at 519.86±3.07, all manifested a significant enzyme inhibitory activity. R. mucronata manifested the highest activity with IC₅₀ at 0.08±1.82 μg mL⁻¹, followed by C. tagal with IC₅₀ at 0.85±1.46 μg mL⁻¹ and B. sensitivum with IC₅₀ at 2.24±1.58 μg mL⁻¹.

Conclusion

This is the first report on the α-glucosidase inhibitory effect of the six Philippine plants; thus, partly defining the mechanism on why these medicinal plants possess antidiabetic properties.     

European medicinal polypores – A modern view on traditional uses

Ethnopharmacological relevance

In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal–spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores.

Materials and methods

All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge.

Results

Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities.

Conclusions

The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

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.