HPLC测定新疆紫草不同部位中8种羟基萘醌的含量

建立同时测定新疆紫草Arnebia euchroma不同部位中左旋紫草素、β-羟基异戊酰紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、去氧紫草素、异丁酰紫草素、β,β'-二甲基丙烯酰阿卡宁和异戊酰紫草素等8种羟基萘醌含量的高效液相色谱法。采用Waters Xbridge C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-0.05%甲酸水溶液(70∶30),检测波长275 nm,流速1mL·min~(-1),柱温30℃。定量结果显示,新疆紫草各部位中主要成分含量差异较大,根皮含量最高,其次是根,再次是茎残基,根木质部和地上部分含量较低。本研究为确定紫草药用部位提供科学依据。     

新疆紫草及其毛状根中萘醌类化合物的研究

目的运用高效液相色谱-电喷雾离子阱液质联用技术(HPLC-ESI-MS/MS)鉴别新疆紫草中的萘醌类化合物,比较分析了9个样品中(新疆紫草原植物根及毛状根)萘醌类成分种类和含量的差异。方法采用BDS C18色谱柱,以0.1%甲酸(A)-甲醇(B)梯度洗脱,使用ESI离子源,流速1.0 m L·min-1,质量扫描范围(m/z)150~400,负离子模式下采集数据。结果新疆紫草中含有紫草素、β-羟基异戊酰紫草素、乙酰紫草素、去氧紫草素、异丁酰紫草素、β,β-二甲基丙烯酰紫草素、异戊酰紫草素或α-甲基-正丁酰紫草素7种萘醌类化合物;但其毛状根中不含紫草素成分,其他成分与新疆紫草相同。紫草素及其衍生物的种类及含量因不同区域、不同毛状根而有所差异。此外,硝酸银(Ag NO3)及营养成分能够增加毛状根中紫草素及其衍生物的种类且影响各成分的含量。结论萘醌类化合物的种类及含量受环境及培养条件的影响而变化。该方法可为新疆紫草及其毛状根的药效物质基础研究及其开发利用提供科学依据。     

HPLC测定新疆紫草中紫草素的含量及提取方法的优化

目的:采用超声方法提取新疆紫草中紫草素,并以左旋紫草素为指标,建立HPLC测定紫草素的含量。方法:利用超声波协同甲醇提取40 min;色谱条件为Shim-pack CLC-ODS-18色谱柱(4.6 mm×250 mm,10μm),检测波长516 nm,流动相甲醇-水(85∶15),流速1.0 mL·min-1,柱温室温。结果:左旋紫草素进样量在0.127~1.143μg具有良好的线性关系,回归方程为Y=338 763X-198 347(r=0.999 8),平均回收率99.3%,RSD 1.18%,左旋紫草素超声法提取率是回流提取法的1.29倍。结论:经方法学验证,该实验所采用的实验方法重复性好、稳定、准确、可靠,可用于中药紫草药材中紫草素的含量测定。     

DNA条形码鉴定与功效组分测定在紫草质量评价中的应用

目的:利用DNA条形码技术对中药紫草进行基源鉴定,并建立紫草的功效多组分含量测定方法,为全面评价紫草药材质量提供依据。方法:对不同产地紫草进行DNA条形码基原鉴定;采用高效液相色谱法测定紫草中萘醌类成分与含量,Thermo BDS Hypersil C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相0.2%冰乙酸-乙腈梯度洗脱,流速0.8 m L·min~(-1),检测波长275 nm,柱温35℃。结果:经DNA条形码鉴定了11批药材,均为紫草科植物新疆紫草Arnebia euchroma的干燥根;测定了11批新疆紫草的8种萘醌类功效组分含量,各组分均显示良好分离;不同成分在线性范围内线性关系良好,平均回收率在95.26%~101.40%,RSD均在0.7%~1.8%(n=6)。结论:DNA条形码鉴定确保了新疆紫草药材基源的准确性,组分含量测定方法准确可靠、重复性好,二者结合可作为紫草的质量控制方法。     

高效液相色谱法测定玉红膏中紫草素的含量

目的采用高效液相色谱测定玉红膏中紫草素的含量。方法选用C18柱,流动相:甲醇-0.05%磷酸水(80∶20),检测波长520 nm,柱温:30℃。结果线性范围为0.22~1.1μg(r=0.999 8),平均回收率99.4%,RSD=1.8%(n=5)。结论此方法可以用于玉红膏中紫草素的含量控制。     

吴紫糊中左旋紫草素的含量测定

目的：建立吴紫糊中左旋紫草素的测定方法。方法：高效液相色谱法,色谱条件色谱柱为安捷伦C18柱（4.6mm×250mm,5μm）,流动相为甲醇-0.1%磷酸盐缓冲液（80∶20）,检测波长516nm,柱温25℃,流速10mL/min。结果：进样量在8.2∽41.0μg/mL范围内,线性关系良好（r=0.999 9）平均回收率为101.30%（RSD为1.55%,n=9）。结论：高效液相色谱法测定吴紫糊中左旋紫草素含量结果准确、重现性好,适用于吴紫糊的质量控制。     

HPLC法测定新疆软紫草属4种植物根中萘醌类成分

目的采用HPLC法测定新疆软紫草属4种植物(新疆紫草、黄花软紫草、天山软紫草、硬萼软紫草)根中7种萘醌类成分(左旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、去氧紫草素、异丁酰紫草素、β,β′-二甲基丙烯酰阿卡宁、2-甲基丁基酰紫草素)的含有量。方法该药材石油醚提取物的分析采用Agilent-C_(18)柱(250 mm×4.6 mm,5μm);流动相乙腈-0.05%甲酸;体积流量1 mL/min;柱温30℃;检测波长275 nm。结果 7种成分在各自范围内线性关系良好,平均加样回收率为97.37%～99.75%,RSD为0.42%～1.94%。结论该方法准确稳定,重复性好,可用于软紫草的质量控制。     

HPLC法测定紫黄凝胶中左旋紫草素的含量

目的:建立以高效液相色谱法测定紫黄凝胶中左旋紫草素的含量.方法:色谱柱为DiamonsilTM(钻石)C<,18>柱(250 mm×4.6 mm,5μm);流动相为甲醇;0.1%磷酸溶液(85;15);柱温为25℃;流速为1.0 mL/min;检测波长为516 nm.结果:HPLC法测定的左旋紫草素含量在0.021 3～0.213 0μg范围内线性关系良好,相关系数r=0.999 8,平均回收率为99.57%,RSD=2.29%.结论:该法操作简便、结果准确,重现性好,可用于本品的控制.     

HPLC法测定紫草油中左旋紫草素的含量

目的建立紫草油中左旋紫草素的含量测定方法.方法采用HPLC法测定紫草油中左旋紫草素的含量,色谱柱:岛津Shim-pack VP-ODS柱(4.6 mm×250 mm);甲醇-0.025 mol/L磷酸(85:15)为流动相;检测波长:516 nm;柱温:25℃;进样量:20 μL.结果左旋紫草素在11.2 μg/mL～33.6 μg/mL浓度范围内线性关系良好(r=0.9998);平均回收率为101.03%,RSD=1.90%(n=5).结论该方法简便、准确,能排除其他成分的干扰,可用于紫草油的质量控制和评价.     

RP-HPLC法测定紫草中左旋紫草素的含量

目的建立HPLC法测定紫草中左旋紫草素含量的方法。方法采用Shimpack C18柱（4.6×250 mm,5μm）,甲醇：0.1%磷酸溶液（85：15）为流动相,流速为1.0 m L·min^-1,柱温：25℃,检测波长：515 nm。结果左旋紫草素浓度在5.3-106μg·m L-1之间与峰面积线性关系良好,r=0.999,8;平均回收率为99.5%,RSD为1.98%。结论本法操作简便,重复性好,可用于紫草中左旋紫草素的质量控制。     

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.     

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.     

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.     

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。     

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。     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Regulation effects on abnormal glucose and lipid metabolism of TZQ-F,a new kind of Traditional Chinese Medicine

Aim of the study

Based on the recipe of the traditional anti-diabetic formula TZQ, we developed TZQ-F, a new formula including 8 fractions isolated from Red Paeony root, Mulberry leaf, Lotus leaf, Danshen root and Hawthorn leaf with a good quality assurance. The study was aimed at fraction preparation and effects of the fractions on abnormal glucose and lipid metabolism.

Materials and methods

The active fractions were obtained by macroporous resin, ion-exchange resin and polyamide resin column chromatographies. HPLC analyses were used for quality control. In vitro mechanism study included DPPH radical scavenging, AGEs formation inhibition, α-glucosidase inhibition and lipase inhibition, and rats on high-fat diet were used for in vivo study.

Results

In vitro mechanism study showed that among the 8 fractions, three of them had inhibition effects on intestinal disaccharase, three with inhibition effects on lipase, and five with effects of free radical scavenging. In vivo study showed that after 4 weeks of treatment, TZQ-F significantly decreased the levels of serum total cholesterol, TG, glucose, LDL-C and HDL-C in rats on high-fat diet. Consistent with the in vitro and in vivo results, histology study demonstrated that TZQ-F alleviated hepatic steatosis induced by high-fat diet.

Conclusions

TZQ-F possesses the potential regulation effects on abnormal glucose and lipid metabolism.     

Antifungal activity of Brazilian medicinal plants involved in popular treatment of mycoses

A survey of medicinal plants used to treat common mycoses was done in the Curituba district, Sergipe State, Brazil. One hundred inhabitants were interviewed by health agents and traditional healers. Four different plants were the most cited (more than 50% of the citations): Ziziphus joazeiro, Caesalpinia pyramidalis, Bumelia sartorum and Hymenea courbaril. The aqueous extracts obtained following traditional methods and using different parts of these plants, were submitted to drop agar diffusion tests for primary antimicrobial screening. Only the water infusion extract of Ziziphus joazeiro and Caesalpinea pyramidalis presented a significant antifungal activity against Trichophyton rubrum, Candida guilliermondii, Candida albicans, Cryptococcus neoformans and Fonsecaea pedrosoi, when compared to the antifungal agent amphotericin B. The minimal inhibitory concentration (MIC) of the bioactive extracts was evaluated by the microdilution method. Best activity with a MIC of 6.5 microg/ml for both extracts was observed against Trichophyton rubrum and Candida guilliermondii. Ziziphus joazeiro and Caesalpinea pyramidalis extracts presented also low acute toxicity in murine models. The present study validates the folk use of these plant extracts and indicates that they can be effective potential candidates for the development of new strategies to treat fungal infections.     

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.     

木蝴蝶种子化学成分研究

目的:研究紫葳科Bignoniaceae植物木蝴蝶Oroxylum indicum种子的化学成分.方法:采用硅胶柱色谱、Sephadex LH-20凝胶柱色谱等手段进行分离纯化,通过化合物的理化常数和波谱数据鉴定其结构.结果:从木蝴蝶种子的95％乙醇提取物中分离并鉴定了20个化合物,分别为木蝴蝶苷A(1)、木蝴蝶苷B(2)、白杨黄素(3)、黄芩苷元(4)、槲皮素(5)、芹菜素(6)、山柰酚(7)、槲皮素-3-O-阿拉伯吡喃糖苷(8)、羽扇豆醇(9)、2α,3β-二羟基羽扇豆醇(10)、赤松素(11)、二氢赤松素(12)、胆甾-5-烯-3,7-二醇(13)、连翘环己醇(14)、异连翘环己醇(15)、zarzissine(16)、(E)-银松素-3-O-β-D-吡喃葡萄糖苷(17)、腺苷(18)、β-谷甾醇(19) 、β-胡萝卜苷(20).结论:化合物11-13,15～18为首次从木蝴蝶植物中分离得到,其中除化合物18外,其余6个化合物均为首次从紫葳科植物中获得.