肉豆蔻制霜新工艺优选及质量评价

目的: 建立机械压榨法制备肉豆蔻霜的新工艺,并与传统工艺进行比较。 方法: 以出油量及性状为指标,采用单因素试验考察压榨次数和温度对机械压榨工艺的影响。以制霜前后脂肪油、挥发油、总木脂素、去氢二异丁香酚的含量为指标,考察传统法与机械压榨法制霜工艺对肉豆蔻中化学成分的影响。采用HPLC测定去氢二异丁香酚含量,色谱条件为Diamonsil C₁₈色谱柱(4.6 mm×250 mm,5 μm),柱温30 ℃,流动相甲醇-水梯度洗脱,检测波长274 nm,流速1.0 mL·min^-1,进样量20 μL。 结果: 优选的机械压榨制霜工艺为压榨温度80~90 ℃,压榨2次,出油率达30.8%。与生品相比,肉豆蔻机械制霜后脂肪油、挥发油、总木脂素、去氢二异丁香酚的含量均降低,但与传统压榨后含量相近。 结论: 机械压榨法方便、快捷,利于工业化生产,可替代传统压榨法。     

山核桃叶总黄酮苷元及其主要单体成分球松素查尔酮的抗氧化活性

目的: 测定山核桃叶总黄酮苷元(TFA)及其主要单体成分——球松素查尔酮(PSC)的抗氧化活性。 方法: 采用柱层析法分离、富集山核桃叶中的总黄酮苷元及单体化合物球松素查尔酮;测定总黄酮苷元中总酚含量,通过1,1-二苯基-2-三硝基苯肼(DPPH)自由基实验、铁离子还原/抗氧化能力(FRAP)测定法、抗脂质过氧化(LPO)实验、抗蛋白氧化羰基(PCO)生成实验,对TFA和PSC进行体外抗氧化活性研究。 结果: 富集的山核桃叶总黄酮苷元中总酚含量用没食子酸当量(GAE)表示为(283.75±10.82)mg·g^-1;TFA和PSC 对DPPH自由基清除的抑制浓度(IC₅₀)分别为(20.05±1.74),(11.66±0.61) mg·L^-1,抑制脂质过氧化的IC₅₀分别为(74.62±5.39),(24.38±1.24) mg·L^-1,抑制蛋白过氧化的IC₅₀分别为(174.29±11.26),(86.74±7.89) mg·L^-1;TFA和PSC均具有较强的抗氧化能力;除了对铁离子还原力外,其他各项指标化合物PSC均显著强于TFA。 结论: 山核桃叶中含有较高含量的黄酮苷元,其富集的总黄酮苷元具有较强的抗氧化活性,已知成分大于50%,而球松素查尔酮是其主要药效物质,值得进一步开发应用。     

HPLC同时测定糖克软胶囊中5种木脂素类成分的含量

 目的建立反相高效液相色谱法同时测定糖克软胶囊中5种木脂素类(五味子醇甲、五味子醇乙、当归酰戈米辛H、五味子甲素、五味子乙素)成分含量的方法。方法色谱条件:Agilent Extend C₁₈柱;流动相为甲醇-2%四氢呋喃水溶液,线性梯度洗脱,流速0.8 mL·min^-1;柱温50℃,检测波长254 nm。结果5种木脂素类化合物在实验浓度内线性关系良好,五味子醇甲、五味子醇乙、当归酰戈米辛H、五味子甲素和五味子乙素的线性范围分别为15.8～1 575,5.76～576,4.65～465,3.61～361,10.6～1 057 mg·L^-1,相关系数均大于0.999 0。平均加样回收率在100.2%～103.5%之间。5种木脂素类化合物含量重复性试验RSD小于1.98%。结论本方法简单、灵敏、重复性好,可用于糖克软胶囊质量控制。     

五味子木脂素与生态因子相关性研究

建立五味子中五味子醇甲、五味子甲素、五味子乙素、五味子丙素4种成分的高效液相色谱测定方法,分析不同样地间气候因子和海拔高度对五味子木脂素类含量的影响,确定吉林省五味子最适栽培地区的气候特点。采用高效液相色谱法,Agilent ZORBAX SB-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相甲醇（A）-水（B）梯度洗脱,检测波长254 nm,柱温 35℃,流速1 mL·min^-1,该测定方法重复性、稳定性和精密度均良好。并用SPSS软件分析气候因子和木脂素相关性,结果发现采集于吉林省集安市、柳河县、安图县和抚松县的五味子成熟果实中五味子醇甲质量分数均高于0.4%,达到质量要求,但是,五味子醇甲、五味子甲素和五味子乙素含量与样地海拔高度间呈明显的线性负相关关系,即随海拔高度增加呈下降趋势,特别是五味子醇甲的负线性拟合系数达到0.844 1,具有显著性,而五味子丙素与样地海拔高度的相关性不明显。与五味子木脂素中五味子醇甲、五味子乙素以及4种木脂素含量呈显著正相关的气候因子主要是温度因子（年均气温、7月最高温、7月平均温、1月最低温、1月平均温）和降水因子（年均降水量）,说明较高的温度和降水量有助于五味子中木脂素的形成和累积。建议五味子栽培区域应尽可能选择在温暖多雨的低海拔地区。     

五味子醋制前后指纹图谱的分析比较

目的: 建立五味子指纹图谱测定方法,分析比较五味子醋制前后物质基础的变化。 方法: 采用HPLC,以乙腈-15 mmol·L^-1磷酸二氢钾溶液(pH 2.0)为流动相,梯度洗脱,检测波长210,254 nm,流速1.0 mL·min^-1,柱温35 ℃,分别测定生、醋五味子指纹图谱。 结果: 五味子醋制前后指纹图谱有明显的差异,其中以5-羟甲基糠醛含量变化最显著。 结论: 建立的方法可以较好体现出五味子醋制前后物质基础的差异,为五味子饮片的质量评价标准提供了科学依据,也为进一步揭示五味子炮制原理的科学内涵奠定了基础。     

三七叶总皂苷脂质体凝胶剂的制备及其离体透皮率考察

目的: 优选三七叶总皂苷脂质体凝胶剂的处方工艺并考察其离体累积透皮吸收率。 方法: 采用薄膜分散法制备脂质体,以包封率为指标,通过正交试验考察选择磷脂与胆固醇的质量比、脂类与药物的质量比、药物质量浓度对三七叶总皂苷脂质体处方工艺的影响。运用高速离心法测定包封率,以卡波姆940为基质制备凝胶剂,通过单因素试验筛选卡波姆940用量。通过体外透皮试验考察三七叶总皂苷中人参皂苷Rb₃的体外透皮吸收情况,利用HPLC测定透过及滞留在皮肤内的人参皂苷Rb₃含量,色谱条件为ZORBOX SB-C₁₈色 谱柱(4.6 mm×250 mm,5 μm),流动相乙腈(A)-0.2% 磷酸(B)梯度洗脱(0~19 min,30% A;19~21 min,35% A;21~26 min,50% A),检测波长203 nm。 结果: 优选的处方工艺为卵磷脂-胆固醇(4:1), 人参皂苷Rb₃质量浓度90 g·L^-1,脂药比(6:1),当卡波姆940质量分数为0.5% 时,药物的皮肤累积透过量最大。 结论: 脂质体凝胶能提高药物中有效成分在皮肤的透过量,为三七叶总皂苷新剂型和新给药途径的开发提供参考。     

HPLC测定家蚕中1-脱氧野尻霉素的含量

目的: 建立RP-HPLC测定家蚕中1-脱氧野尻霉素(DNJ)的含量。 方法: 采用Hypersil NH₂色谱柱(4.6 mm×250 mm,5 μm),以乙腈-水(78:22)为流动相,流速1.0 mL·min^-1,柱温30 ℃,检测波长203 nm。 结果: 家蚕中DNJ在1.996~9.98 μg (r=0.999 6)峰面积与进样量呈良好的线性关系,平均加样回收率为98.4%。 结论: 该方法前处理简单、实用,结果准确,可用于家蚕及制品中DNJ的质量控制。     

胶束电动毛细管电泳法快速分离测定南、北五味子中木脂素类成分

目的：建立胶束电动毛细管电泳法测定五味子中6种木脂素类成分的方法。方法：探讨了缓冲溶液、添加剂、分离电压、温度和进样条件等因素对分离检测的影响。在10 mmol·L^-1 NaH₂PO₄-Na₂HPO₄缓冲溶液（35%乙腈,37.5 mmol·L^-1 SDS,pH 8.0）、分离电压为28.0 kV的优化条件下,9 min内可实现五味子样品溶液的分析。结果：线性范围分别为25.5~1 020,25.75~1 030,12.0~480,25.5~1 020,24.25~970,19.75~790 mg·L^-1;检出限为0.5,0.8,1.0,1.0,1.0,1.5 mg·L^-1。结论：该方法与高效液相色谱法比较,在分离效果、柱效和分析速度方面具有明显优势。     

HPLC测定小活络丸(浓缩丸)中11-羰基-β-乙酰乳香酸

目的: 建立小活络丸(浓缩丸)中11-羰基-β-乙酰乳香酸的高效液相色谱含量测定方法。 方法: Inertsil ODS-3 C₁₈(4.6 mm×250 mm,5 μm)色谱柱,流动相乙腈-0.1%冰醋酸 (80:20),流速1 mL·min^-1,检测波长249 nm,柱温30 ℃,外标法测定小活络丸(浓缩丸)中乳香主成分11-羰基-β-乙酰乳香酸。 结果: 11-羰基-β-乙酰乳香酸在25.95~207.6 mg·L^-1线性关系良好(R²=0.999 9);平均加样回收率为99.95%,RSD 3.29% (n=6)。 结论: 方法操作简便,结果准确,重复性良好,可用于小活络丸(浓缩丸)中乳香的质量控制。     

ARPD精制液中4个指标成分的定量测定

目的: 建立抗放射性肺炎方(anti-radiation pneumonia decoction, ARPD)精制液中4个指标成分的定量分析方法。 方法: 以芍药苷、阿魏酸、咖啡酸、甘草酸为考察指标,采用岛津LC-20AD系列高效液相色谱仪和Kromasil C₁₈色谱柱(4.6 mm×250 mm,5 μm),流动相乙腈-0.1% 磷酸,进行梯度洗脱,流速1.0 mL·min^-1,柱温30 ℃。 结果: ARPD精制液中芍药苷、阿魏酸、咖啡酸和甘草酸的质量浓度分别为522,13,12,143 mg·L^-1。 结论: 各指标成分检测方法的稳定性、准确度和回收率良好,可用于ARPD精制液的质量控制。     

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。     

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.     

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.     

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.     

虫草属EST-SSR标记系统的建立研究

目的:通过球孢虫草、蛹虫草EST设计EST-SSR引物,建立虫草属EST-SSR标记系统.方法:从NCBI公共数据库下载获得虫草EST,利用Sequece Seiners 1.2软件去除冗余序列并设计引物,进行PAGE电泳.结果:通过去除EST总序列中低质量的和冗余的序列后,得到全长为2 953 173 bp的4 556条无冗余球孢虫草EST.从中发掘出718个EST-SSR,分布于616条EST中,出现频率是15.8％.平均分布频率是每4 096 bp出现1个,三核苷酸重复序列有419个,是出现最多的重复类型.蛹虫草EST去冗余后得到1 363条无冗余EST,共含有1 117个EST-SSR,出现频率为81.95％,出现最多的重复类型是A核苷酸重复.根据球孢虫草EST-SSR序列,设计合成50对引物,有扩增产物的引物为34对,占总设计引物数的68％.根据蛹虫草EST-SSR,设计合成40对引物,有扩增产物的引物为39对,占总设计引物数的97.5％.基于SSR标记进行聚类分析,7种虫草无性型均能分开,且分为4支.结论:虫草属EST-SSR出现频率较高、类型较丰富、多态性潜能较高,具有较高的利用价值.球孢虫草和蛹虫草EST开发的SSR标记在虫草属有较好的转移性与通用性,可以很好的应用于虫草种间遗传关系的研究.应用虫草物种EST建立分子标记是一条简便而又有效的途径.     

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。