升麻及其炮制品的HPLC指纹图谱研究

目的 RP-HPLC法建立升麻指纹图谱.方法 采用Phenomsil BDS C18(250 mm ×4.6 mm,5μm)色谱柱;流动相为乙腈(A)-0.1％磷酸水溶液(B)梯度洗脱,0～5 min(88％A:12％B),25 min(80％A:20％B),45 min(70％A:30％B),60min(60％A:40％B),检测波长316 nm,柱温40℃;体积流量1.0 mL/min;进样量10μL.结果 建立了升麻的HPLC 指纹图谱,标定出15个共有指纹峰.结论 升麻药材的指纹图谱可用于升麻的定性鉴别,为升麻药材的质量控制提供参考依据.     

元胡止痛片HPLC指纹图谱研究

目的建立元胡止痛片HPLC指纹图谱。方法采用Extent C18柱(250 mm×4.6 mm,5μm),流动相为0.1%磷酸溶液(三乙胺调pH 6.5,A)-甲醇(B),梯度洗脱程序为:0～45 min,88%～40%A;45～70 min,40%～20%A;70～75 min,20%～10%A;75～80 min,10%～5%A;80～85 min,5%A;检测波长280 nm,柱温35℃,体积流量1.0 mL/min。结果在HPLC色谱指纹图谱中,确定了16个共有峰,建立了元胡止痛片的共有模式,在14批元胡止痛片样品中有10批样品的指纹图谱相似度在0.9以上。结论本法结果准确、重现性好,为元胡止痛片HPLC指纹图谱质量控制提供了参考。     

五灵脂药材的HPLC指纹图谱分析

目的:建立五灵脂药材的HPLC指纹图谱.方法:采用HPLC法,色谱柱Diamonsil ODS C18 (4.6 mm ×250 mm,5 μm);流动相乙腈(A)-0.5％冰醋酸水溶液(B);梯度洗脱(0～ 15 min,85％ ～ 65％B,15～35 min,65％ ～ 40％B,35 ～ 40min,40％～25％B,40～ 60 min,25％ ～ 10％B,60 ～65 min,10％～0％B;检测波长265 nm;流速0.8 mL· min-1;柱温室温.结果:建立了五灵脂药材的HPLC指纹图谱,标定了14个共有峰,并用对照品指认了其中3个峰,10批样品的相似度均大于0.90.方法的精密度、稳定性和重复性良好.结论:建立的五灵脂药材指纹图谱特征性及专属性强,可为其质量控制和药效物质基础研究提供参考.     

宫血宁胶囊HPLC指纹图谱研究

目的 本研究建立宫血宁胶囊(重楼)的HPLC指纹图谱分析方法,为宫血宁胶囊内在质量评价积累数据.方法 采用RP-HPLC方法,以Agilent Series SB-AQ C1s色谱柱(4.6 mm×250 mm,5 μm);流动相为乙腈(A)-水(B),梯度洗脱程序为5～50 min,20％～42％A;50～70 min,42％～44％A;70～100 min,44％～80％A;100～120 min,80％～95％A;洗脱时间为120 min;检测波长203 nm;体积流量1.0 mL/min;柱温30℃;进样量10μL,进行指纹图谱研究.结果 建立宫血宁胶囊指纹图谱,以重楼皂苷H色谱峰为参照峰,确定12个共有峰,测定了60批样品,样品指纹图谱相似度均大于0.99.结论 本法操作简便、分离效果好、稳定性高、重现性好,可应用于宫血宁胶囊的质量控制及真伪品鉴别.     

广西不同产区两面针HPLC指纹图谱研究

目的 采用高效液相色谱法建立两面针的HPLC色谱指纹图谱分析方法,为其品质控制提供可靠依据.方法 采用HPLC-UV分析两面针的指纹图谱.色谱柱:UltimateTMXB C8柱(250 nn×4.6 mm,5μm);流动相为水(0.2％磷酸+0.2％三乙胺)(A)-乙腈(B),梯度洗脱;0～5 min,8％～12％ B; 5～7 main,12％～15％B;7～14 main,15％～21％ B; 14～35min,21％～25％B; 35～40min,25％～36％B; 40～60min,36％～52％B; 60～80min,52％～100％B; 80～95min,100％B;体积流量1.0 mL/min柱温35℃;检测波长250 nm,测定24批两面针指纹图谱,应用相似度分析、系统聚类分析,对两面针进行分类研究.结果 在色谱指纹图谱中,确定了22个共有峰,根据相似度分析、系统聚类分析的结果,将24批药材分为2类.结论 该指纹图谱检测方法方便,重现性好,可用于两面针质量控制.     

回心草药材HPLC指纹图谱研究

目的 建立回心草药材的指纹图谱.方法 采用反相高效液相色谱,色谱条件为:Dikma Platisil ODS C18柱(250 mm×4.6 mm,5 μm);流动相A:0.5%醋酸乙腈,B:0.5%醋酸水溶液;洗脱方法:0～50 min,A为20%～55%;50～60 min,A为55%～95%,60～85 min,A为95%～100%,保持5 min;体积流量1.0 mL/min,波长360nm.柱温:30℃,运用中药色谱指纹图谱相似度评价系统2004版(国家药典委员会)进行分析.结果 构建了回心草药材的指纹图谱.结论 该法准确.重现性好,可作为回心草药材的质控方法.     

维吾尔药材菠菜子的DNA条形码鉴定及质量控制研究

目的:基于DNA条形码分子鉴定法建立维吾尔药材菠菜子的鉴定方法,并构建菠菜子HPLC指纹图谱和其指标性成分β-蜕皮甾酮的HPLC含量测定方法,为菠菜子的质量控制提供参考。方法:以核糖体DNA第二内部转录间隔区(ITS2)为主体条形码序列对菠菜子进行DNA条形码鉴定;采用Agilent ZORBAX SB-Aq C_(18)色谱柱(4.6 mm×250mm,5μm),以甲醇(A)-0.2%磷酸溶液(B)为流动相梯度洗脱(0～10 min,10%A; 10～20 min,10%→30%A;20～40 min,30%→40%A; 40～65 min,40%→80%A),检测波长240 nm,建立菠菜子的HPLC指纹图谱方法;采用Agilent ZORBAX SB-Aq C_(18)色谱柱(4.6 mm×250 mm,5μm),以甲醇(A)-0.2%磷酸溶液(B)为流动相梯度洗脱(0～15 min,45%→40%A; 15～25 min,40%→80%A; 25～30 min,80%→45%A; 30～45 min,45%A),检测波长248 nm,建立指标性成分β-蜕皮甾酮的HPLC含量测定方法。结果:DNA条形码鉴定系统进化树显示,ITS2序列可将菠菜子与其近缘物种很好区分,且分支支持率达100%; HPLC指纹图谱标定7个共有峰,10批菠菜子药材与对照图谱的相似度均大于0.95;指标性成分β-蜕皮甾酮在6.115～122.304μg/m L范围内线性关系良好(r=0.9997),重复性试验RSD 2.0%,加样回收率为97.43%～103.59%,10批样品中β-蜕皮甾酮的含量为0.099%～0.241%。结论:DNA条形码可准确鉴定菠菜子的基原,HPLC指纹图谱专属性强,结合指标性成分β-蜕皮甾酮的含量测定可以控制菠菜子药材的质量。     

小金丸脂溶性和水溶性部位的HPLC指纹图谱分析

目的:建立小金丸脂溶性部位、水溶性部位的高效液相色谱法(HPLC)指纹图谱并进行指纹图谱相似度评价,探讨小金丸的质量一致性。方法:采用Welch Ultimate AQ-C18色谱柱(4.6 mm×250 mm,5μm),流动相选择0.1%磷酸水溶液(A)-乙腈(B)梯度洗脱(脂溶性部分:0～5 min,40%B;5～10 min,40%～50%B;10～20 min,50%～60%B;20～30 min,60%～65%B;30～40 min,65%～70%B;40～50 min,70%～80%B;50～60 min,80%～90%B;60～65 min,90%～95%B;65～75 min,95%～100%B;75～80 min,100%B。水溶性部分:0～20 min,2%～5%B;20～30 min,5%～10%B;30～37 min,10%～20%B;37～45 min,20%～30%B;45～50 min,30%～40%B;50～58 min,40%B),流速1 mL·min~(-1),柱温30℃,脂溶性部位和水溶性部位的检测波长分别设定为202,250 nm,进样量依次为10,20μL。对5个厂家共30批市售小金丸进行HPLC检测,使用主成分分析(PCA)对各部位色谱峰进行分析,并对色谱峰进行指认。结果:指纹图谱共检测出55个色谱峰,30批小金丸指纹图谱相似度差异较大。小金丸脂溶性部位、水溶性部位的指纹图谱相似度RSD分别为21.5%和32.8%,厂家间与厂家内样品质量差异较大,且脂溶性部位成分主导小金丸化学一致性评价结果,其中,来自枫香脂的成分是影响该制剂化学差异的主要原因。结论:市售小金丸质量一致性较差。双部位指纹图谱的建立为小金丸整体质量评控提供了新思路,可为传统丸剂的质量一致性评价提供参考。     

丹参波长叠加指纹图谱及多指标成分测定研究

对丹参波长叠加指纹图谱和7种丹参指标成分同时定量进行研究。采用Agilent Eclipse Plus C_(18)(4. 6 mm×100 mm,3. 5μm)色谱柱,以0. 1%甲酸的水溶液(A)-乙腈(B)为流动相梯度洗脱(0～5 min,10%～20%B; 5～20 min,20%～30%B; 20～25 min,30%～50%B; 25～40 min,50%～65%B; 40～45 min,65%～80%B; 45～46 min,80%～10%B; 46～50 min,10%B),流速1mL·min~(-1),柱温30℃,检测波长250,280,310,340 nm。运用波长叠加技术建立指纹图谱,并选取主要成分建立多指标测定方法。波长叠加指纹图谱体现了丹参250,280,310,340 nm特征吸收波长的指纹信息,17个不同来源的丹参样品与指纹图谱之间具有良好的相似性,相似度在0. 828～0. 936。确定了15个共有峰,并指认了其中7个色谱峰,分别是迷迭香酸、丹酚酸B、丹酚酸A、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA,其方法学考察结果良好。建立了丹参波长叠加谱指纹图谱,该方法简便可靠、耐用性好,可用于丹参的质量控制。     

不同产地白术饮片HPLC指纹图谱研究

目的:通过对不同产地白术饮片研究，建立白术饮片高效液相指纹图谱。方法:采用HPLC法。色谱柱：Agilent C₁₈(5μm, 250 mm×4.6 mm);流动相：乙腈-水，梯度洗脱(0～10 min, 50%～70%A;10～20 min, 70%～80%A,20～30 min; 80%～92%A,30～38 min, 92%～100%A),流速为1.0mL·min^-1,柱温为25℃,白术内酯Ⅰ、白术内酯Ⅲ检测波长为220 nm,白术内酯Ⅱ检测波长为276 nm,进样量20μL。以2号峰白术内酯Ⅲ为参照，记录12批次白术饮片的HPLC图谱，然后将其导入《中药色谱指纹图谱相似度评价软件系统2012版》(国家药典委员会),采用指纹图软件确定共有峰。结果:建立12批白术饮片的HPLC指纹图谱，标定12个共有峰，相似度均大于0.90。经验证，与对照指纹图谱具有较好的一致性。结论:研究建立HPLC指纹图谱可用于白术饮片质量控制方法，同时也为白术饮片质量评价提供科学依据。     

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.     

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.     

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。     

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.     

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.     

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.     

贝母类药材湖北贝母Fritillaria hupehensis系统位置的探讨——来自ITS,rpl16,matK序列的证据

湖北贝母为传统中药,然而《Flora of China》将其基原植物湖北贝母Fritillaria hupehensis归并于天目贝母F.monantha项下。该实验采用分子系统学方法,以川百合Lilium davidii为外类群,用核基因ITS序列和叶绿体基因rpl16序列、matK序列等3个片段对湖北贝母及其近缘类群天目贝母F.monantha、安徽贝母F.anhuiensis等进行联合建树分析,对湖北贝母植物的系统位置进行了探讨,为湖北贝母药材的安全使用提供分子证据。结果显示,分子系统树上,3种贝母各自的居群聚为一支,之后天目贝母与安徽贝母聚为一支,最后与湖北贝母聚为一支。表明湖北贝母与天目贝母的亲缘关系可能要远于安徽贝母与天目贝母之间的关系,因此不适宜将湖北贝母归并于天目贝母。     

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。