HPLC法测定印度獐牙菜中芒果苷的含量

目的:测定印度獐牙菜中芒果苷的含量。方法:用高效液相色谱法测定芒果苷的含量,选择ZORBAXSB-C18(4.6mm×150mm,5μm)柱;流动相组成:甲醇-1%磷酸二氢钠-四氢呋喃(25∶75∶1)(磷酸调pH至3.2);检测波长λ=265nm。结果:测定芒果苷的平均回收率为96.91%,芒果苷含量为4.476mg/g。结论:本方法可有效用于印度獐牙菜中芒果苷的测定。     

藏药材印度獐牙菜质量标准研究

 目的 建立印度獐牙菜药材的质量标准。方法 采用薄层色谱法鉴别印度獐牙菜药材中獐牙菜苦苷、龙胆苦苷和芒果苷成分;以高效液相色谱法测定该3种成分的含量。色谱柱为Zorbax Eclipse XDB-C₁₈（4.6 mm×250 mm,5 μm）,流动相为甲醇-水（0.1%冰醋酸）线性梯度洗脱,甲醇体积分数从22%经18 min至32% 并保持5 min,流速为1.0 mL·min-1,柱温为25 ℃,检测波长为243 nm。结果 印度獐牙菜药材中獐牙菜苦苷、龙胆苦苷和芒果苷的薄层色谱鉴别特征明显,专属性强;药材中獐牙菜苦苷、龙胆苦苷和芒果苷的含量测定线性范围分别为0.06~2.00 μg(r=0.999 7),0.09~2.90 μg(r=0.999 7)和0.05~1.65 μg(r=0.999 8),平均回收率（n=9）分别为103.76 % (RSD=1.34 %),99.43 % (RSD=1.60%)和96.22% (RSD=1.30%)。结论 所建立的印度獐牙菜定性定量测定方法简单准确,能够为印度獐牙菜药材的质量控制提供有效数据。     

川东獐牙菜植物器官獐牙菜苦苷含量分布规律研究

目的:研究獐牙菜苦苷在川东獐牙菜各植物器官中的含量分布规律。方法:反相高效液相色谱法(RP-HPLC),以Kramasil C_(18)柱(150mm×4.6mm,5μm)为分析柱,以甲醇-水(30:70)为流动相,柱温25℃,流速0.8ml/min,检测波长为240nm。结果:牙菜苦苷进样量在1.317～11.854μg范围内与峰面积有良好的线性关系,r=0.9996,平均加样回收率为98.92%,RSD=0.59%。结论:獐牙菜苦苷在川东獐牙菜各器官中的含量分布规律是花叶全草茎枝。     

川东獐牙菜不同部位芒果苷的含量测定研究

目的建立高效液相色谱法测定川东獐牙菜不同部位芒果苷的含量。方法采用Hypersil C18柱(150 mm×4.6mm,5μm),流动相为甲醇-0.04%磷酸溶液在0-15 min内由22∶78到32∶68,流速1.0 ml.min-1,检测波长254 nm,柱温室温,用外标法定量,测定川东獐牙菜中芒果苷的含量。结果芒果苷的线性范围0.27-2.70μg,r=0.999 7,回收率100.90%,RSD为2.19%。结论该方法简便,快速,线性关系良好,可用于川东獐牙菜芒果苷的含量测定。     

不同海拔川西獐牙菜中药用成分的HPLC分析

目的:建立藏药材川西獐牙菜中四种药用成分的HPLC测定方法,比较不同海拔川西獐牙菜中四种主要药用成分的含量变化。方法:采用高效液相色谱法测定四种成分的含量,测定三种苷类成分采用甲醇-水为流动相,进行梯度洗脱,检测波长为260 nm,柱温为25℃;测定齐墩果酸采用甲醇-含0.04%冰乙酸水为流动相,90%甲醇等度洗脱,检测波长为210 nm,柱温为30℃。结果:实验所建立的测定方法简便稳定;在2 100～3 617 m的不同海拔地区所采集的川西獐牙菜中獐牙菜苦苷、龙胆苦苷、芒果苷和齐墩果酸的含量变化范围分别是1.82～3.13、27.17～34.61、9.45～25.53、2.91～7.08 mg/g。结论:不同海拔地区的藏药材川西獐牙菜中,齐墩果酸含量随海拔增高而显著增加,龙胆苦苷含量随海拔增高明显降低,獐牙菜苦苷含量随海拔增高而略有增加,芒果苷含量随海拔增高呈现出增加的趋势,但趋势变化有较大的波动。     

藏药川西獐牙菜的质量标准研究

目的:建立川西獐牙莱药材的质量标准.方法:采用薄层色谱法对川西獐牙菜药材进行定性鉴别,采用高效液相色谱法测定主要活性成分獐牙莱苦苷、芒果苷、当药醇苷和齐墩果酸的含量.色谱柱为Diamonsil C18(4.6 mm ×250 mm,5μm),检测獐芽菜苦苷、芒果苷和当药醇苷的流动相甲醇-0.02％磷酸水(45:55),检测波长254 nm,检测齐墩果酸的流动相乙腈-甲醇-0.2％冰醋酸水(65:15:20),检测波长210 nm,流速1.0 mL·min-1,柱温30℃.结果:川西獐牙菜药材的薄层色谱鉴别特征明显,专属性强;獐牙菜苦苷、芒果苷、当药醇苷和齐墩果酸的HPLC含量测定线性范围分别为0.075～0.75μg(r=0.999 9),0.05 ～0.5 μg(r=0.999 7),0.032～0.32 μg(r=0.999 3),0.116～11.6 μg(r=0.999 4),平均回收率分别为98.91％ (RSD 0.77％),99.61％ (RSD1.38％),100.35％ (RSD 0.69％),99.26％(RSD 0.40％),并根据10批药材中相应成分的含量初步制定了其含量限度.结论:川西獐牙菜定性定量测定方法简单准确,能够为川西獐牙菜药材的质量控制提供有效依据.     

藏药印度獐牙菜提取工艺研究

目的：确定印度獐牙菜药材最佳渗漉提取工艺。方法：以HPLC法测定芒果苷含量及出膏率为指标,采用平行比较试验法,优选印度獐牙菜药材的提取方式和提取工艺条件。结果：印度獐牙菜药材最佳提取工艺为采用75%乙醇渗漉法提取,以4 mL/min/kg的流速,收集药材6倍量渗漉液。结论：优选得到的提取工艺简便易行,稳定性好。     

HPLC法同时测定青叶胆中獐牙菜苦苷、龙胆苦苷和獐牙菜苷的含量

目的:建立青叶胆中獐牙菜苦苷、龙胆苦苷、獐牙菜苷的含量测定方法,为控制其质量提供科学依据。方法:采用高效液相色谱法。色谱条件:色谱柱为Phenomsil C18色谱柱(250 mm×4.6 mm,5μm);流动相为甲醇-0.02%磷酸溶液(23∶77);检测波长为243 nm;流速为1 mL/min;柱温为30℃。结果:3种成分均达到基线分离,獐牙菜苦苷、龙胆苦苷、獐牙菜苷的线性范围分别为0.33～1.98μg(r=0.9997),0.16～0.96μg(r=0.9999),0.20～1.20μg(r=0.9997),回收率分别为100.2%,99.5%,98.2%。结论:该法快速、准确、稳定,可用于青叶胆药材质量评价。     

UPLC法同时测定藏药复方八味獐牙菜丸中獐牙菜苦苷、龙胆苦苷和芒果苷的含量

目的:以藏药复方八味獐牙菜菜丸为研究对象,建立活性成分獐牙菜苦苷、龙胆苦苷、芒果苷的定量分析方法。方法:采用UPLC法,色谱柱为Waters Acquity UPLC C18(2.1mm×100mm,1.7μm);流动相为甲醇-0.02%磷酸溶液(梯度洗脱);体积流量0.2mL/min;检测波长254nm。结果:獐牙菜苦苷、龙胆苦苷、芒果苷在25～333μg的质量浓度范围内与峰面积呈良好的线性关系(r2=0.9997、r2=0.9992、r2=0.9994);平均回收率分别为95.50%、102.93%、93.76%,RSD分别为3.84%、4.09%、2.75%。结论:所建立的UPLC法简便、快捷、准确,可用于复方八味獐牙菜丸活性成分的定量测定及质量控制,为安全、合理、有效用药及制定药材质量标准提供科学依据。     

高效液相法同时测定獐牙菜属植物中四种活性成分

目的：同时测定獐牙菜属植物中四种抗肝炎活性成分的含量。方法：用高效液相色谱法对獐牙菜苦苷、芒果苷、当药醇苷和雏菊叶龙胆酮四种抗肝炎活性成分进行了分析。结果：在选定的色谱条件下四种成分分离良好,在检测浓度范围内线性良好。日内和日间精密度分别小于2%和4.6%,加样回收率为97.94%～99.84%。建立了獐牙菜属植物中四种不同化学结构类型抗肝炎活性成分的含量测定方法。结论：本方法简单、准确,能成功的的运用于同时测定獐牙菜属植物不同部位中獐牙菜苦苷、芒果苷、当药醇苷和雏菊叶龙胆酮四种抗肝炎活性成分的含量以及獐牙菜属植物的鉴定。     

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.     

Antitubercular activity of Arctium lappa and Tussilago farfara extracts and constituents

Ethnopharmacological relevance

Arctium lappa and Tussilago farfara (Asteraceae) are two plant species used traditionally as antitubercular remedies. The aim of this study was (i) to screen Arctium lappa and Tussilago farfara extracts for activity against Mycobacterium tuberculosis and (ii) to isolate and identify the compound(s) responsible for this reputed anti-TB effect.

Materials and methods

The activity of extracts and isolated compounds was determined against Mycobacterium tuberculosis H₃₇R_v using a high throughput spot culture growth inhibition (HT-SPOTi) assay.

Results

The n-hexane extracts of both plants, the ethyl acetate extract of Tussilago farfara and the dichloromethane phase derived from the methanol extract of Arctium lappa displayed antitubercular activity (MIC 62.5 μg/mL). Further chemical investigation of Arctium lappa led to the isolation of n-nonacosane (1), taraxasterol acetate (2), taraxasterol (3), a (1:1) mixture of β sitosterol/stigmasterol (4), isololiolide (5), melitensin (6), trans-caffeic acid (7), kaempferol (8), quercetin (9), kaempferol-3-O-glucoside (10). Compounds isolated from Tussilago farfara were identified as a (1:1) mixture of β sitosterol/stigmasterol (4), trans-caffeic acid (7), kaempferol (8), quercetin (9), kaempferol-3-O-glucoside (10), loliolide (11), a (4:1) mixture of p-coumaric acid/4-hydroxybenzoic acid (12), p-coumaric acid (13). All compounds were identified following analyses of their physicochemical and spectroscopic data (MS, ¹H and ¹³C-NMR) and by comparison with published data. This is the first report of the isolation of n-nonacosane (1), isololiolide (5), melitensin (6) and kaempferol-3-O-glucoside (10) from Arctium lappa, and of loliolide (11) from Tussilago farfara. Amongst the isolated compounds, the best activity was observed for p-coumaric acid (13) (MIC 31.3 μg/mL or 190.9 μM) alone and in mixture with 4-hydroxybenzoic acid (12) (MIC 62.5 μg/mL).

Conclusions

The above results provide for the first time some scientific evidence to support, to some extent, the ethno-medicinal use of Arctium lappa and Tussilago farfara as traditional antitubercular remedies.     

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.     

In vitro and in vivo antimalarial and cytotoxic activity of five plants used in congolese traditional medicine

Aim of the study

The in vitro antiplasmodial activity and cytotoxicity of methanolic and dichloromethane extracts from five Congolese plants were evaluated. The plants were selected following an ethnobotanical survey conducted in D.R. Congo and focusing on plants used traditionally to treat malaria. The in vivo antimalarial activity of aqueous and methanolic extracts active in vitro was also determined in mice infected by Plasmodium berghei berghei.

Materials and methods

The growth inhibition of Plasmodium falciparum strains was evaluated using the measurement of lactate dehydrogenase activity. The extracts (aqueous, CH₃OH, EtOH and CH₂Cl₂) were prepared by maceration and tested in vitro against the 3D7 (chloroquine sensitive) and W2 (chloroquine resistant) strains of Plasmodium falciparum and against the human normal fetal lung fibroblasts WI-38 to determine the selectivity index. Some extracts were also used at the dose of 300 mg/kg to evaluate their activity in mice infected since 4 days by Plasmodium berghei.

Results

Two plants presented a very high activity (IC₅₀ < 3 μg/ml). These plants were Strychnos icaja roots bark (MeOH and CH₂Cl₂) and Physalis angulata leaves (MeOH and CH₂Cl₂). One plant (Anisopappus chinensis whole plant, MeOH and CH₂Cl₂) presented a high activity (IC50 < 15 μg/ml). The extracts of Anisopappus chinensis and Physalis angulata showed also a good inhibition of parasitemia in vivo. Flavonoids, phenolic acids and terpenes were identified in these plants by a general phytochemical screening method.

Conclusion

Three plants showed a very interesting antiplasmodial activity (Anisopappus chinensis, Physalis angulata and Strychnos icaja) and one of them showed a good selectivity index (>10, Anisopappus chinensis). Anisopappus chinensis and Physalis angulata were also active in vivo.     

牛耳朵和黄花牛耳朵的显微和化学鉴别

目的:为苦苣苔科唇柱苣苔属植物牛耳朵和黄花牛耳朵的鉴定和分类提供解剖学和化学依据。方法:采用石蜡制片法和水合氯醛透化法对2种药用植物的根状茎和叶横切面和粉末特征进行研究,应用光学显微镜观察显微结构。采用HPLC-UV法进行化学鉴别。结果:牛耳朵和黄花牛耳朵显微特征无明显区别,但是化学特征有明显的差异。结论:化学诞生特征鉴别方法可以作为2种药用植物的鉴定方法。     

HPLC-DAD同时测定柘树和构棘根与茎中4种黄酮类成分的含量

目的:分析比较柘树与构棘根及茎中花旗松素、柑桔素、槲皮素、山柰酚的含量,为品种鉴定与质量评价提供科学参考.方法:采用高效液相二极管阵列检测法(HPLC-DAD),色谱柱为Agilent C18(4.6 mm×150 mm,5μm),流动相为甲醇(A)-0.1％磷酸溶液梯度洗脱(0 ～ 15 min,35％ ～50％ A;15 ～30 min,50％ ～65％A),检测波长290 nm(花旗松素、柑桔素)和365 nm(槲皮素、山柰酚),柱温30℃.结果:花旗松素、槲皮素的含量柘树根明显高于构棘根,柘树茎明显高于构棘茎,种间含量差异大;柑桔素、山柰酚的含量构棘根高于柘树根,柘树茎与构棘茎含量相当,种间含量差异小,同种药用部位间含量差异大.结论:4种成分的含量在柘树和构棘及其药用部位根和茎都存在一定的差异,在临床上不可混淆使用.     

Dragon's blood: botany, chemistry and therapeutic uses

Dragon's blood is one of the renowned traditional medicines used in different cultures of world. It has got several therapeutic uses: haemostatic, antidiarrhetic, antiulcer, antimicrobial, antiviral, wound healing, antitumor, anti-inflammatory, antioxidant, etc. Besides these medicinal applications, it is used as a coloring material, varnish and also has got applications in folk magic. These red saps and resins are derived from a number of disparate taxa. Despite its wide uses, little research has been done to know about its true source, quality control and clinical applications. In this review, we have tried to overview different sources of Dragon's blood, its source wise chemical constituents and therapeutic uses. As well as, a little attempt has been done to review the techniques used for its quality control and safety.     

不同类型的桑螵蛸与其基原昆虫对应关系研究

桑螵蛸为常用中药,因其入药部位为螳螂的卵鞘,无法直接判断其基原昆虫.同时螳亚科动物种类繁多,均产卵鞘,目前关于桑螵蛸对应的基原昆虫尚无一致的结论,而二者准确的对应关系是保证桑螵蛸质量的基础,也是桑螵蛸条形码研究的基础.文章采用人工孵化与捕捉活体螳螂2种方式,分别鉴定了药材及其原动物,系统地研究了桑螵蛸与基原昆虫的对应关系,发现基原昆虫为大刀螳属、斧螳属、污斑螳属的多种昆虫,该研究为桑螵蛸深入开发与利用研究奠定了基础.     

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。     

Sesquiterpene lactones from Gynoxys verrucosa and their anti-MRSA activity

Ethnopharmacological relevance

Because of its virulence and antibiotic resistance, Staphylococcus aureus is a more formidable pathogen now than at any time since the pre-antibiotic era. In an effort to identify and develop novel antimicrobial agents with activity against this pathogen, we have examined Gynoxys verrucosa Wedd (Asteraceae), an herb used in traditional medicine in southern Ecuador for the treatment and healing of wounds.

Materials and methods

The sesquiterpene lactones leucodine (1) and dehydroleucodine (2) were extracted and purified from the aerial parts of Gynoxys verrucosa, and their structure was elucidated by spectroscopic methods and single-crystal X-ray analysis. The in vitro anti-microbial activity of Gynoxys verrucosa extracts and its purified constituents was determined against six clinical isolates including Staphylococcus aureus and Staphylococcus epidermidis strains with different drug-resistance profiles, using the microtiter broth method.

Results

Compound 1 has very low activity, while compound 2 has moderate activity with MIC₅₀s between 49 and 195 μg/mL. The extract of Gynoxys verrucosa has weak activity with MIC₅₀s between 908 and 3290 μg/mL.

Conclusions

We are reporting the full assignment of the ¹H NMR and ¹³C NMR of both compounds, and the crystal structure of compound 2, for the first time. Moreover, the fact that compound 2 has antimicrobial activity and compound 1 does not, demonstrates that the exocyclic conjugated methylene in the lactone ring is essential for the antimicrobial activity of these sesquiterpene lactones. However, the weak activity observed for the plant extracts, does not explain the use of Gynoxys verrucosa in traditional medicine for the treatment of wounds and skin infections.