一测多评法在玄参药材质量控制中的应用

目的建立玄参药材中哈巴苷、类叶升麻苷、安格洛苷C、哈巴俄苷、肉桂酸5种成分的一测多评法,验证该法在玄参质量评价中的准确性和可行性。方法以哈巴苷为内参物,分别建立肉桂酸、类叶升麻苷、安格洛苷C、哈巴俄苷的相对校正因子(f),并计算其含量,实现一测多评。与外标法所得结果进行比较,对一测多评价法的可行性进行验证。结果建立的相对校正因子分别为f_(哈巴苷/肉桂酸)=0.060(RSD=0.81%)、f_(哈巴苷/类叶升麻苷)=0.068(RSD=0.53%)、f_(哈巴苷/安格洛苷C)=0.197(RSD=1.82%)、f_(哈巴苷/哈巴俄苷)=0.142(RSD=1.17%),差异性较小;25批玄参中肉桂酸、类叶升麻苷、安格洛苷C、哈巴俄苷含量的实测值与计算值无显著差异。结论一测多评法控制玄参药材的质量是准确、可行的。     

HPLC-UV波长转换法测定玄参药材及饮片中哈巴苷与哈巴俄苷的含量

目的:建立同时测定中药玄参中哈巴苷与哈巴俄苷的HPLC-UV双波长含量测定方法,考察炮制对2种成分含量的影响,提出玄参药材和饮片中哈巴苷和哈巴俄苷的含量限度建议.方法:应用Agilent Technologies ZORBAX SB- C18 (4.6mm× 250 mm,5μm)色谱柱,以乙腈-0.03％磷酸水溶液为流动相,进行梯度洗脱,流速1.0 mL·min-1,柱温为25℃,采用双波长检测13 min前用210 nm,13 min以后用280 nm作为检测波长.结果:哈巴苷和哈巴俄苷能够达到很好的分离.哈巴苷线性范围为0.0549～1.46 μg;哈巴俄苷线性范围为0.022 5 ～0.900 μg.哈巴苷与哈巴俄苷平均回收率分别为98.1％,RSD2.4％(n=9);98.8％,RSD 4.3％ (n =9).10批玄参商品药材中含量哈巴苷为0.277％～0.620％,哈巴俄苷为0.078％～0.362％;10批玄参商品饮片中含量哈巴苷为0.276％～1.059％,哈巴俄苷为0.059％～0.183％;即哈巴苷平均含量玄参饮片(0.567％)高于药材(0.448％),哈巴俄苷平均含量饮片(0.128％)低于药材(0.237％).而同批次玄参药材在自制加工成饮片后哈巴苷含量值升高13.7％～ 96.0％,哈巴俄苷含量值降低11.0％～ 73.9％.结论:所建立的含量方法操作简便,结果准确,可用于玄参质量控制.玄参药材加工成饮片的过程可使哈巴苷含量值升高、哈巴俄苷含量值降低.建议玄参药材及饮片均以其中哈巴苷和哈巴俄苷总含量以干燥品计算应不低于0.45％为质量标准.     

HPLC同时检测增液汤中5种成分的含量

目的:建立HPLC法同时测定增液汤中5种成分含量的方法。方法:采用ZORBAX SB C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.1%乙酸溶液,梯度洗脱;流速:1.0 mL/min;柱温:30℃;进样量:20μL,测定7批增液汤中安格洛苷C、哈巴俄苷、肉桂酸、哈巴苷、麦角甾苷5种成分的含量。结果:哈巴苷、麦角甾苷、安格洛苷C、哈巴俄苷、肉桂酸分别在0.036～0.426 mg/mL(r=0.999)、0.003～0.036 mg/mL(r=0.999)、0.012～0.141 mg/mL(r=0.999)、0.012～0.138 mg/mL(r=0.999)和0.003～0.039 mg/mL(r=0.998)范围内与峰面积呈良好的线性关系,平均加样回收率分别为103.76%、104.17%、97.52%、106.52%和113.68%,RSD分别为3.74%、4.00%、4.92%、5.40%和4.70%。结论:该方法简便、准确且重复性好,可为增液汤的质量控制提供参考。     

不同配比苍术-玄参药对中4种成分溶出率的变化规律

目的采用HPLC法研究不同配比苍术-玄参药对中哈巴苷、哈巴俄苷、安格洛苷C、苍术素溶出率的变化规律。方法分析采用Phenomenex Luna C_(18)色谱柱(250 mm×4.6 mm,5.0μm);流动相为乙腈-水,梯度洗脱;体积流量1.0 m L/min;检测波长210 nm(哈巴苷)、280 nm(哈巴俄苷、安格洛苷C、苍术素);柱温25℃。结果当药对配伍比例为2∶1时,4种成分的总溶出率最高,而为1∶2时最低。结论哈巴苷和安格洛苷C的溶出率随苍术比例的增加而升高,而哈巴苷的溶出率和哈巴俄苷可能成反比关系。     

玄参对照提取物研究及其在玄参药材质量控制中的应用

建立玄参对照提取物制备工艺并进行标定,探讨其替代相应对照品在玄参药材质量控制中应用的可行性.采用溶剂提取法和色谱分离技术制备玄参对照提取物,以哈巴苷、安格洛苷C和哈巴俄苷对照品为对照,对玄参对照提取物进行含量标定.分别以已知含量的玄参对照提取物和哈巴苷、安格洛苷C、哈巴俄苷3个对照品为对照,建立玄参药材HPLC含量测定...     

高效液相色谱-紫外双波长法同时测定玄参配方颗粒中3种主要成分的含量

目的建立高效液相色谱-紫外双波长法同时测定玄参配方颗粒中3种主要成分哈巴苷、哈巴俄苷和肉桂酸含量的方法。方法采用Ultimate AQ-C18色谱柱(250 mm×4.6 mm,5μm),流动相为1%冰醋酸水溶液和乙腈,梯度洗脱,流速1.0 mL/min,检测波长前13 min为210 nm,13 min后为278 nm,柱温30℃。结果哈巴苷、哈巴俄苷、肉桂酸的线性范围分别为0.066 54~0.665 4μg(r=1.000 0)、0.024 23~0.242 3μg(r=0.999 9)、0.100 28~1.002 8μg(r=0.999 9),平均加样回收率(n=6)分别为99.80%±1.22%、100.31%±1.30%、100.22%±1.24%。结论本法简单、灵敏度高、重复性好、准确可靠,可为玄参配方颗粒的质量控制以及标准研究提供参考依据。     

HPLC测定增液承气口服液中哈巴苷和哈巴俄苷含量

目的:建立增液承气口服液中哈巴苷和哈巴俄苷的含量测定方法.方法:采用高效液相色谱法,色谱柱为Eclipse XDB C18柱(4.6 mm×150 mm,5μm),以乙腈-0.03％磷酸溶液为流动相进行梯度洗脱,检测波长210 nm,柱温30℃,流速1.0 mL? min-1.结果:哈巴苷和哈巴俄苷进样量分别在0.257 ～1.284 μg(r=0.999 8),0.092 ～0.460 μg(r=1.000 0)时,与峰面积值具有良好的线性关系,平均加样回收率分别为98.74％( RSD 0.58％),97.15％ (RSD 1.02％).结论:样品处理简便、结果准确、重复性好,可用于增液承气口服液内在质量的控制方法.     

基于指纹图谱结合多指标测定的玄参药材等级质量研究

目的建立玄参药材的HPLC指纹图谱,并同时进行多成分含量测定,为玄参药材等级质量标准的制定提供方法和依据。方法采用HPLC-DAD法,Elipse XDB-C18(250 mm×4.6 mm,5μm)色谱柱,柱温25℃,以乙腈(A)-0.03%磷酸水溶液(B)为流动相进行梯度洗脱,体积流量1.0 mL/min,进样量10μL,指纹图谱检测波长207 nm,含量测定的检测波长为哈巴苷210 nm、哈巴俄苷280 nm、肉桂酸264 nm。获取30批不同等级玄参药材的指纹图谱,采用中药色谱指纹图谱相似度评价系统(2012版)进行分析,同时对3种主要化学成分进行含量测定。结果指纹图谱分析共指认7个共有峰,相对保留时间RSD值均小于2.0%,相对峰面积RSD值差异较大,提示玄参药材中的主要化学成分稳定存在,但含量差异较大;相似度分析中,约有5%的指纹图谱与对照药材的相似度低于0.7,12.5%的相似度在0.7～0.8,40%处于0.8～0.9,42.5%大于0.9,提示不同产地玄参药材的质量差异较大;不同批次玄参药材中3种化学成分含量差异较大,质量分数分别为哈巴苷0.18%～2.89%、哈巴俄苷0.01%～0.35%、肉桂酸0.01%～0.24%。结论建议在原有等级划分标准的基础上,增加指纹图谱和主要化学成分的含量测定两项指标。     

HPLC-UV-ELSD同时测定复方红景天胶囊中8种成分

目的采用HPLC-UV-ELSD同时测定复方红景天胶囊中大花红天素、没食子酸、红景天苷、酪醇、哈巴苷、哈巴俄苷、安格洛苷C、肉桂酸8种成分。方法应用Kromasi L色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.3%冰醋酸水溶液,梯度洗脱,体积流量1.0 m L/min,紫外检测波长275 nm,柱温30℃,蒸发光散射检测器漂移管温度40℃,载气N2,压力150 k Pa。结果大花红天素、没食子酸、红景天苷、酪醇、哈巴苷、哈巴俄苷、安格洛苷C、肉桂酸8种成分能够达到很好分离;其线性范围分别为67.084~670.84(R~2=0.999 2)、11.410~114.100(R~2=0.999 4)、78.995~789.95(R~2=0.999 6)、19.625~196.25(R~2=0.999 7)、59.368~593.68(R~2=0.999 8)、62.585~625.85(R~2=0.999 5)、55.045~550.45(R~2=0.999 6)、6.895~68.95μg/m L(R~2=0.999 8);平均回收率分别为100.8%、98.9%、100.1%、100.8%、98.9%、99.6%、100.7%、99.2%,RSD分别为0.64%、0.56%、0.35%、0.65%、0.26%、0.58%、1.00%、0.64%(n=6)。结论该方法分析简便、准确,可作为该制剂的定量分析方法。     

甲肿消制剂及其含药血清中哈巴俄苷的含量测定

目的 建立甲肿消制剂及灌胃给予甲肿消后大鼠血清中的哈巴俄苷含量测定方法.方法 采用RP-HPLC法测定哈巴俄苷的含量,色谱柱:Kromasil C_(18)色谱柱(250 mm×4.6 mm,5 μm)和Easyguard保护柱,以乙腈和1%醋酸溶液为流动相,采用梯度洗脱;流速:1 ml· min~(-1);检测波长为278 nm;进样量:5 μl.结果 哈巴俄苷在1.46～146 μg·ml~(-1)的浓度范围内线性良好;低、中、高浓度哈巴俄苷的精密度(RSD)(n=5)分别为1.05%,0.59%,0.80%;日间差(RSD)(n=5)为1.00%;制剂和大鼠血清中哈巴俄苷的加样回收率(n=5)分别为(98.02±1.01)%和(101.03±1.64)%;测定其重现性(RSD)(n=6)分别为0.73%和0.96%;每克制剂中含哈巴俄苷的量为230.5 μg,每毫升含药血清中哈巴俄苷的量为1.88 μg.结论 该方法简便、快速、准确、重现性良好,可用于制剂和血清中哈巴俄苷的含量测定.     

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。     

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.     

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。     

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.     

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。