元胡止痛片中重金属及铅、镉、汞的测定

目的:测定元胡止痛片(元胡,白芷)中重金属总量及铅、镉、汞的含量.方法:中国药典2000版一部附录重金属检查法及对外贸易经济合作部颁布的<药用植物及制剂进出口绿色行业标准>中有关铅、镉、汞的测定法.结果:元胡止痛片中重金属总量符合要求,但铅、镉、汞含量均有不同程度的超标.结论:仅用重金属总量不足以保证药品质量,尚需建立有害元素铅镉汞等的含量限度.     

中药房常用饮片重金属含量测定

目的：测定10种中药饮片中5种重金属的含量。方法：参照《药用植物及制剂进出口绿色行业标准》，按照《中国食品卫生理化检验手册》1997版检测，用石墨炉原子吸收仪等仪器测定铅、镉；用原子吸收仪检测镍；用测汞仪检测汞；砷斑法检测砷。结果：35个样本中有18个样本重金属含量超标，10种中药饮片中有7种重金属含量超标。     

沙枣中重金属镉含量分析

目的：建立一种用于测定沙枣中重金属镉含量的方法。方法：采用硝酸-高氯酸（4：1）消解法处理不同产地的沙枣样品,用石墨炉原子吸收分光光度法测定镉含量。结果：14批药材均检出一定量的镉,但含量较低,未超出《药用植物及制剂进出口绿色行业标准》中对重金属镉的规定量（Cd≤0.3 mg.Kg-1）。测定方法的加标回收率为99.68%,相对标准偏差为0.79%。结论：该试验方法操作简便、快速,具有良好的精密度和准确度,可用于沙枣药材中重金属元素镉含量的测定。     

松杉灵芝29种微量元素分析

目的：分析松杉灵芝微量元素含量,为松杉灵芝用药安全提供依据。方法：采用电感耦合等离子体质谱仪测定松杉灵芝中29种微量元素的含量。结果：松杉灵芝菌盖中重金属元素Pb含量为（4.21±0.29）μg·g^-1,Cu含量为（12.55±1.12）μg·g^-1,As含量为（1.26±0.08）μg·g^-1,Cd未检出。松杉灵芝菌柄中重金属元素Pb含量为（3.63±0.04）μg·g^-1,Cu含量为（16.59±1.24）μg·g^-1,As含量为（1.58±0.05）μg·g^-1,Cd未检出。松杉灵芝菌盖中Al,Fe,Zn,Mn含量比赤芝菌盖高。结论：松杉灵芝中重金属元素Pb<5 μg·g^-1,Cd<0.3 μg·g^-1,Cu<20 μg·g^-1,As<2.0 μg·g^-1,符合药用植物及制剂进出口绿色行业标准的限量标准。     

闽产47种中成药中重金属的含量测定

 目的　以福州、漳州、厦门、建阳等 4个地区的47种中成药为研究对象进行铅、砷、汞、镉含量测定。方法　按国标食品中重金属含量的测定方法。结果　参照药用植物及制剂进出口绿色行业标准 ,超标率(%)为:铅0;镉0;汞53.23;砷2.13;结论　铅、镉的含量在福建省的几个地区含量不高,符合标准,个别样品含砷量超标,汞的项目超标率较高。     

国槐不同组织器官中多成分分析及抗氧化活性研

目的 通过对国槐Sophora japonica不同组织器官进行综合评价，为其资源合理利用提供依据。方法 采用高效液相色谱（HPLC）法、电感耦合等离子体发射光谱（ICP-OES）法、比色法检测样品中芦丁、水仙苷、槲皮素、异鼠李素、重金属含量以及Fe³⁺还原能力、DPPH自由基（DPPH·）、ABTS自由基（ABTS^·+）清除能力和总抗氧化能力（FRAP）。运用SPSS 20.0软件对结果进行聚类分析和主成分分析。结果 国槐不同组织器官中4种黄酮类成分总含量为花蕾 > 花 > 花轴 > 叶 > 枝条；抗氧化能力依次为花蕾 > 花 > 花轴 > 叶 > 枝条，与黄酮类成分总量呈正相关。花和花蕾中5种重金属含量均符合《药用植物及制剂进出口绿色行业标准》，部分叶、花轴和枝条中Cd超标。聚类分析将花和花蕾聚为一类，花轴、叶和枝条聚为另一类；主成分分析将8个变量提取为2个主成分（PC1和PC2），不同组织器官间PC1值有明显差异，不同批次间表现在PC2值上有差异。结论 国槐花及花蕾中重金属含量低，黄酮类成分含量高，抗氧化活性强，品质优良；花轴及叶也含有较高的黄酮类成分及较强的抗氧化能力，具有进一步开发利用的价值。     

不同产地羌活药材中的重金属及砷盐含量研究

目的:对四川、青海和甘肃主产区羌活药材的重金属及砷盐含量进行分析评价,为制定羌活药材重金属含量质量标准提供参考依据。方法:对各主产区19个点57个样的羌活和宽叶羌活药材样品中铜、镉、铅、砷、铬含量(ICP-AES)及汞含量(冷原子吸收法)进行分析。结果:汞未检出;铅在羌活中含量极低,在宽叶羌活中仅个别产地检出较高;铜含量较高,在羌活中为21～73 mg.kg^-1,在宽叶羌活中为26～73 mg.kg^-1;砷在宽叶羌活中含量较高;镉在羌活中为0～0.29 mg.kg^-1,在宽叶羌活中为0～0.32 mg.kg^-1;铬在羌活中为0.48～8.36 mg.kg-1,在宽叶羌活中为1.44～8.02 mg.kg-1,不同产地间有较大差异。结论:羌活药材基本上达到了《药用植物及制剂进出口绿色行业标准》相应的重金属及砷盐限量标准,但铜的含量超标,故应选择土壤和环境中铜含量低的土地进行羌活的规范化种植。     

甘肃五大宗药材农残及重金属检测

目的:参照中国药典及有关资料对甘肃五大宗药材(当归、党参、大黄、黄芪、红芪)中有机氯类农药及重金属残留量进行检测.方法:采用气相色谱法测定农残留量,采用标准加入法,用石墨炉原子吸收法测定砷、镉、铅、铜的含量,用冷原子吸收法测定汞含量.结果:上述五大宗药材中有机氯类农药残留量均符合<药用植物及制剂进出口绿色行业标准>(中华人民共和国对外贸易经济合作部)规定.重金属残留量除个别产地的品种外,其余均符合规定.     

“宛药”唐半夏中重金属的含量测定

目的:考察唐半夏药材重金属的含量。方法:用微波消解技术处理药材,重金属Cu、Pb、Cd采用石墨炉法测定;Hg、As采用原子荧光法测定。结果:唐半夏药材中重金属的含量较低,符合药典和《药用植物及制剂进出口绿色行业标准》要求。结论:本研究完善了唐半夏的质量标准,为其GAP建设提供理论支撑。     

黔产艾纳香对土壤中重金属的吸收富集特征研究

目的 为使中药材艾纳香的种植栽培科学化,以贵州省罗甸艾纳香种植基地为研究区域,从地球化学角度研究艾纳香对土壤重金属吸收富集特征.方法 以<药用植物及制剂进出口绿色行业标准>的限定值为标准,对重金属含量进行评价.结果 重金属已存在超标现象.结论 艾纳香对不同重金属元素的吸收富集能力有较大的差异,整体呈现出对Cd的富集能力＞对Cu的富集能力＞对Pb的富集能力的变化趋势.相关性分析表明,艾纳香植株对重金属元素吸收过程中,Cd和Cu这两种重金属元素之间可能存在竞争性机制.     

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.     

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.     

Scorpio and Scolopendra attenuate inflammation and articular damage in rats with collagen-induced arthritis

Ethnopharmacological relevance

Scorpio and Scolopendra (SS) are two traditional Chinese medicines, which are generally used to treat rheumatoid arthritis (RA) in China. However, the mechanism is so far unclear. The purpose of this study was to explore the effects and mechanisms of SS in attenuating inflammation and joint injury in collagen-induced arthritis in rats.

Materials and methods

RA was induced in Wistar rats by injection of collagen, meanwhile, the rats were administrated daily either SS (0.4 g/kg, 0.2 g/kg, and 0.1 g/kg) or vehicle (physiological saline) for 42 days. The therapeutic effect of SS on RA was evaluated by pathological methods. T lymphocyte subsets and anti-collagen type II (CII) antibody were tested in peripheral blood. Tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4) and interleukin-10 (IL-10) were assessed in tissue homogenate of fresh joints.

Results

The inflammation and articular damage in SS powder-treated rats were attenuated significantly. In addition, SS powder was revealed to modulate the equilibrium of T lymphocyte subsets, down-regulate TNF-α and IL-1β, up-regulate IL-4 and IL-10, and significantly suppress the level of anti-CII antibody.

Conclusions

Scorpio and Scolopendra, when used as a combination, reveal desirable effect for treatment of rheumatoid arthritis, and this beneficial effect may be accomplished through normalization of T lymphocyte subsets and the balance of Th1/Th2 cytokines.     

In vitro antitrypanosomal and antileishmanial activity of plants used in Benin in traditional medicine and bio-guided fractionation of the most active extract

Ethnopharmacological relevance

The aim of the study was to evaluate the in vitro antitrypanosomal and antileishmanial activity of crude extracts of 10 plant species traditionally used in Benin to treat parasitic infections.

Materials and methods

For each species, dichloromethane, methanol and aqueous extracts were tested. Their antitrypanosomal and antileishmanial activities were evaluated in vitro on Trypanosoma brucei brucei (strain 427) (Tbb) and on promastigotes of Leishmania mexicana mexicana (MHOM/BZ/84/BEL46) (Lmm).

Results

The best growth inhibition was observed with the dichloromethane extracts of aerial parts of Acanthospermum hispidum DC. (Asteraceae) (IC₅₀ = 14.5 μg/ml on Tbb and 11.1 μg/ml on Lmm), twigs of Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (IC₅₀ = 5.8 μg/ml on Tbb), aerial parts of Byrsocarpus coccineus Schumach. & Thonn (syn. Rourea coccinea (Schumach. & Thonn.) Hook.f.) (IC₅₀ = 14.7 μg/ml on Tbb) and aerial parts of Carpolobia lutea G.Don. (IC₅₀ = 18.3 μg/ml on Tbb). All these extracts had a low cytotoxicity. It is not the case for the methanolic and water extracts of roots of Anchomanes difformis (Blume) Engl. (IC₅₀ = 14.7 and 13.8 μg/ml on Tbb) which were toxic at the same concentration range on WI38, human cells. A bio-guided fractionation of the most active extract of Keetia leucantha allowed to identify oleanolic acid and ursolic acid as responsible for the observed activities.

Conclusion

Our study gives some justification for antiparasitic activity of some investigated plants.     

野生与栽培防风中升麻苷和5-O-甲基维斯阿米醇苷的含量测定及指纹图谱分析

目的:建立并比较野生和栽培品种防风中升麻苷、5-O-甲基维斯阿米醇苷HPLC测定方法和HPLC指纹图谱。方法:含量测定采用岛津C18色谱柱(4.6 mm×150 mm,5μm),流动相为甲醇-水(40∶60),检测波长254 nm,流速1.0 mL.min-1,柱温30℃。HPLC指纹图谱测定采用资生堂MGⅡC18(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-水,梯度洗脱,检测波长254 nm,流速1.0 mL.min-1,柱温30℃。结果:升麻苷加样回收率为99.6%,RSD 0.72%(n=6);5-O-甲基维斯阿米醇苷加样回收率为102.6%,RSD 0.88%。野生与栽培品种升麻苷质量分数分别为(4.96±2.59),(3.61±1.82)mg.g-1;5-O-甲基维斯阿米醇苷质量分数分别为(3.91±2.09),(4.37±2.02)mg.g-1。野生与栽培品种升麻苷和5-O-甲基维斯阿米醇苷质量分数无显著性差异。在此条件下所建立的指纹图谱中防风各成分得到有效分离。结论:升麻苷、5-O-甲基维斯阿米醇苷HPLC测定方法简便,结果准确,HPLC指纹图谱分析可作为防风药材质量控制及分类评价的依据。     

Comparisons on enhancing the immunity of fresh and dry Cordyceps militaris in vivo and in vitro

Ethnopharmacological relevance

The immunomodulatory capacities of fresh Cordyceps militaris (FCM) and dry Cordyceps militaris (DCM) were compared.

Materials and methods

In vivo immunomodulatory assay, different doses of FCM and DCM were orally administrated over a period of 15 days in a cyclophosphamide (CY) induced immunosuppression mice; in vitro testing, the spleen cells were extracted from healthy mice and treaded with CY, then cultured with different dose of FCM or DCM; the contents of Cordyceps militaris polysaccharide (CMP), cordycepin, adenosine, total polyphenol (TP) and total flavonoids (TF) in FCM and DCM were measured.

Results

Our studys indicated that, FCM was significantly stronger than DCM on increasing the spleen and thymus indexes, spleen lymphocyte activity, macrophage function, and promoting the levels of IL-2, IFN-γ in vivo and in vitro. The contents of immunomodulatory CMP and TF in FCM were markedly higher than in DCM.

Conclusion

All these results suggested that FCM was superior to DCM on enhancing immunity.     

四川藏羌地区五加属主要资源种类的HPLC 指纹图谱特征和种类鉴定

目的:建立四川藏羌地区五加属主要资源种类的HPLC指纹图谱,为鉴别其种类提供依据.方法:采用梯度洗脱方法,以Kromasil C18色谱柱(4.6 mm×150 mm,5μm),柱温30℃,以甲醇-0.5％磷酸水溶液为流动相.建立四川藏羌地区五加属药材的指纹图谱,并采用“中药色谱指纹图谱相似度评价系统”软件进行相似度的计算.结果:建立的方法重现性较好,并根据检测结果确定了12个共有特征峰.应用DPSv3.0统计软件对获得的HPLC数量化特征进行系统聚类分析(组间距离法,欧氏距离),聚类分析结果为S1与S2,S3红毛五加药材的不同药用部位,成分最为接近;其次是S5(糙叶藤五加茎皮);再其次分别是S4(蜀五加茎皮)、S8(香加皮药材)、S6(刺五加药材)和S7(五加皮药材).结论:所建立的HPLC指纹图谱对四川藏羌地区五加属主要资源种类的鉴别具有参考价值.     

常用藏药“蒂达”(藏茵陈)基原物种药用合理性及资源利用价值评价

目的:对“蒂达”各品种不同基原植物药用合理性及其资源利用价值作出评价.方法:采用HPLC指纹图谱共有特征峰、结合SPSS聚类分析方法,评价“蒂达”不同品种基原植物的成分组成相似性.结果:“蒂达”不同品种涉及的来源有龙胆科獐牙菜属、花锚属、扁蕾属及肋柱花属共15种1变种33个样品,指纹图谱全谱相似度较差;以10个共有峰成分的相对面积聚类,其基原植物可分别聚为4个组;上述4个属的属间成分组成差异较大;其中獐牙菜属不同种间成分组成变化较大,而其他各属种间成分组成相似度较高.结论:上述龙胆科4属植物均含有具有保肝利胆生物活性的环烯醚萜类、(山)酮类及三萜类成分,藏医学将其归为“蒂达”一类药材使用具有一定的物质基础;鉴于“蒂达”类药材不同基原各品种间在成分组成上差异较大,分别制定各品种的HPLC指纹图谱,是鉴别和控制各品种质量的有效手段,可有效控制各基原品种质量.     

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.