黄花蒿中青蒿素的超临界CO2流体提取工艺研究

研究了用超临界ＣＯ２流体萃取法从黄花蒿中提取青蒿素的工艺,主要探讨了压力,时间、ＣＯ２流量等因素对产品收率的影响,确定了最佳工艺条件,并进行了中试放大和工业化,所得产品质量达药品标准,与传统生产工艺相比,收率提高了１．９倍,生产周期缩短近１００ｈ,生产成本降低了４４７例／ｋｇ。     

UV法与HPLC法测定青蒿中青蒿素含量的比较

目的:分别以高效液相色谱(HPLC)法与紫外分光光度(UV)法测定青蒿中青蒿素的含量,比较2种方法分析结果的差异。方法:HPLC法中色谱柱为Kromasil ODS C18(250mm×4.6mm,5μm),流动相为甲醇-水(75:25),流速为1.0mL·min-1,检测波长为203nm,柱温为30℃。UV法中样品溶液用0.2%NaOH衍生后,在292nm波长处测定。结果:HPLC法标准曲线的线性范围为0.025～0.400mg·mL-1(r=0.9999),平均加样回收率为97.18%(RSD=3.58%,n=9);UV法标准曲线的线性范围为4～20μg·mL-1(r=0.9998)。UV法含量测定结果比HPLC法平均高出2.05倍。结论:HPLC法的精密度、稳定性、加样回收率都较好,对青蒿中青蒿素含量的分析较为准确。UV法不宜单独用于青蒿中青蒿素的含量分析。     

The Effect of Polyphenols Isolated from Cynanchi wilfordii Radix with Anti-inflammatory,Antioxidant, and Anti-bacterial Activity

Recently, Cynanchi wilfordii Radix has gained wide use in Asian countries as a functional food effective for relieving fatigue, osteoporosis, and constipation, particularly in menopausal disorders. However, its anti-inflammatory and anti-microbial activities have not been explored in detail to date. The anti-inflammatory, antioxidant, and anti-bacterial properties of the Cynanchi wilfordii Radix extracts obtained with water, methanol, ethanol, and acetone were compared. All 4 polyphenol-containing extracts exhibited anti-inflammatory and antioxidant effects. The ethanol extract was found to elicit the most potent reduction of nitric oxide (NO), prostaglandin E₂ (PGE₂), and cytokine (IL-1β, IL-6, IL-10, and TNF-α) levels, as well as inhibit the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a concentration-dependent manner. The evaluation of antioxidant activity also revealed the ethanol extract to have the highest free radical scavenging activity, measured as 85.3±0.4%, which is equivalent to 99.9% of the activity of α -tocopherol. In the assessment of anti-bacterial activity, only ethanol extract was found to inhibit the growth of the Bacillus species Bacillus cereus and Bacillus anthracis. These results show that polyphenols of Cynanchi wilfordii Radix have anti-inflammatory, antioxidant, and anti-bacterial properties that can be exploited and further improved for use as a supplementary functional food, in cosmetics, and for pharmaceutical purposes.     

纤维素酶辅助提取黄花蒿中青蒿素的工艺条件研究

目的 研究纤维素酶辅助提取青蒿素反应中各条件对提取率的影响.方法 采用紫外分光光度法测定青蒿素含量,研究酶反应过程中时间、加酶量、温度和pH值对提取率的影响,并使用正交实验方法对提取工艺进行优化.结果 纤维素酶辅助提取青蒿素的最优工艺条件分别是:酶反应时间2h,纤维素酶用量0.30 g,酶反应温度45℃,酶反应pH值4.5.结论 纤维素酶辅助提取青蒿素的方法能够有效提高提取率.     

青蒿素检测方法的研究近况

目的：分析近年来抗疟药物——青蒿素测定方法的研究进展。方法：根据国内外文献资料进行归纳、分析，加以综述。结果：根据青蒿素的理化性质，青蒿素的测定方法已由定性的光谱测定，发展到精确定量的色谱方法。灵敏的电化学和酶联免疫方法也得到发展。结论：准确定量测定青蒿素含量，对于青蒿素生产和药理研究具有重要的意义。     

青蒿素高含量地区黄花蒿种质资源的简单重复序列区间分析

目的:分析广西、贵州和重庆3个青蒿素高含量地区的野生黄花蒿资源的遗传多样性。方法:以40份黄花蒿为试材,通过简单重复序列区间(ISSR)分析,运用POPGENE version1.31软件和TREECONW软件计算相关参数,UPGMA方法聚类,构建亲缘关系树状图。结果:18条ISSR引物扩增出84条带,多态比率为79.76%;物种水平上,ISSR标记揭示的Nei's基因多样性指数为0.2019,Shannon信息指数为0.3244;3个地区之间ISSR标记揭示的Nei's基因多样性指数范围为0.1867～0.1943,Shannon信息指数范围为0.2936～0.3073;ISSR检测不同材料间遗传距离范围为0.1467～0.4493,平均为0.3132。结论:ISSR标记能有效地揭示青蒿素高含量地区野生黄花蒿极为丰富的遗传多样性,可为进一步的选择育种提供更多的种质资源。     

黄花蒿中青蒿素的超临界CO2流体提取工艺研究

研究了用超临界CO2流体萃取法从黄花蒿中提取青蒿素的工艺,主要探讨了压力、时间、CO2流量等因素对产品收率的影响,确定了最佳工艺条件,并进行了中试放大和工业化试验,所得产品质量达药品标准,与传统生产工艺(如汽油法)相比,收率提高了1.9倍,生产周期缩短近100h,生产成本降低了447元/kg。     

岗松总黄酮抗氧化及抗炎作用研究

目的:研究岗松总黄酮体内抗氧化和抗炎作用.方法:小鼠灌胃岗松总黄酮9 d后,测定血清 SOD、MAD、GSH-Px水平以探讨岗松总黄酮的体内抗氧化作用;通过测定岗松总黄酮对二甲苯致小鼠耳肿胀抑制率的影响,研究岗松总黄酮的抗炎作用.结果:岗松总黄酮80 mg·kg-1组可显著提高小鼠SOD、GSH-Px活力,明显降低MAD含量(P＜0.05);而且能显著抑制二甲苯致小鼠耳的肿胀(P＜0.05).结论:岗松总黄酮具有较好的抗氧化和抗炎作用.     

响应面法优化纤维素酶辅助提取青蒿素的工艺研究

目的优化纤维素酶提高青蒿素提取率的反应条件。方法应用响应面实验方法采用中心组合实验设计研究酶反应过程中时间、加酶量、温度对提取率的影响,并对提取工艺条件进行优化。结果纤维素酶辅助提取青蒿素的最优工艺条件：酶反应时间2．9h,酶添加量0．20g,反应温度42．6℃。结论响应面试验方法能够有效提高纤维素酶辅助提取青蒿素的提取率。     

Artemisinin and sesquiterpene precursors in dead and green leaves of Artemisia annua L. crops

This paper analyses the accumulation and concentrations of the antimalarial artemisinin in green and dead leaves of Artemisia annua crops in two field experiments. Concentration differences were analysed as being determined by (a) the total production of artemisinin plus its upstream precursors dihydroartemisinic acid, dihydroartemisinic aldehyde, artemisinic aldehyde and artemisinic alcohol and (b) the conversion of precursors towards artemisinin. Concentrations of the total of artemisinin plus its precursors were higher in green leaves than in dead leaves in the younger crop stages, but were comparable at the final harvests. In every crop stage, the conversion of precursors to artemisinin was more advanced in dead leaves than in green leaves. This resulted in the molar concentrations of artemisinin being higher in dead leaves than in green leaves at the final harvests. The molar quantity of dihydroartemisinic acid, the last enzymatically produced precursor, was higher than that of artemisinin in green leaves, but only 19 - 27% of that of artemisinin in dead leaves. Dead leaves were very important for the final artemisinin yield. They constituted on average 34% of the total leaf dry matter and 47% of the total artemisinin yield at the final harvests. The possibility to convert a larger part of dihydroartemisinic acid into artemisinin during post-harvest handling is discussed.     

不同产地的青蒿药用成分含量研究

目的：研究不同产地的青蒿药用成分的含量。方法：从全国各地采集22批青蒿药材,通过高效液相色谱法测定药材中的青蒿酸、青蒿乙素、青蒿素和东莨菪内酯的含量,并以所测含量为指标,进行相关分析和聚类分析。结果：相关分析和聚类分析均显示,不同产地的青蒿中上述4种成分含量差异较大,各自聚为一类。结论：青蒿活性成分的积累与产地有一定的相关性。     

青蒿研究进展

青蒿有很高的药用价值,应用前景十分广阔。鉴于此,本文从青蒿的主要化学成分、药理学作用及临床应用、生物学特性、组织培养、栽培技术、采收与加工等方面对青蒿近年来的研究进展做一综述,为青蒿的研究与开发提供理论依据。     

红鱼眼醇提物镇痛抗炎作用的实验研究

目的:研究红鱼眼醇提物镇痛、抗炎的作用。方法:采用热板法和醋酸扭体法观察红鱼眼醇提物对小鼠的镇痛作用;采用二甲苯致小鼠耳廓肿胀法和角叉菜胶诱导大鼠足趾肿胀法观察红鱼眼醇提物的抗炎作用。结果:高、中、低剂量红鱼眼醇提物(30、20、10g·kg-1)均能在不同程度上提高小鼠的痛阈值,并能显著抑制小鼠耳廓肿胀和足趾肿胀反应;高剂量红鱼眼醇提物能显著减少大鼠角叉菜胶性炎性渗出液中前列腺素E2含量。结论:红鱼眼醇提物具有一定的镇痛、抗炎作用。     

体外胁迫促进青蒿素在黄花蒿培养细胞中合成的研究

在黄花蒿细胞悬浮培养合成青蒿素的过程中添加微生物刺激剂和微生物胸外酶刺激剂,测定刺激剂对青蒿素合成的影响。采用的微生物刺激剂为霉菌ＡＳ３４,３０９和ＡＳ３,３４６、酵母Ｋluyberomyces fragilis及细菌ＡＳ１,３９８的提取物,微生物胸外酶刺激剂为果胶酶、纤维素酶及半纤维素酶。其中酵母Ｋluyberomyces fragilis提取物和果胶酶对青蒿素生物合成具有明显的刺激作用。用酵母Ｋluyberomyces fragilis提取物（２０％）处理黄花蒿培养细胞３天,青蒿素合成量达到每克黄花蒿干细胞３２０μｇ,比未刺激细胞的青蒿素合成量提高３８％;将果胶酶（０.２％）添加到青蒿素合成培养基中处理黄花蒿培养细胞２天以上,青蒿素合成量达到每克黄花蒿干细胞７４０μｇ,比未刺激细胞的青蒿素合成量提高３.０８倍。     

黄花蒿中青蒿素的微波辅助提取

采用微波辅助提取法提取黄花蒿中的青蒿素。对提取溶剂乙醇、三氯甲烷、环己烷、正己烷、石油醚（30－60℃和60－90℃两种）、120号溶剂油和6号抽提溶剂油进行了比较，考察了溶剂的介电常数对青蒿素得率的影响。并将微波辅助提取法同索氏提取、超临界CO2提取以及加热搅拌提取法进行了比较。     

益生汤抗氧化活性研究

目的:研究益生汤抗氧化作用.方法:用体外化学模拟体系研究了益生汤清除DPPH自由基、羟基(·OH)自由基和超氧阴离子(O2-)自由基的能力以及益生汤总的还原能力、抑制亚油酸过氧化作用进行了测定;通过D-半乳糖诱导的衰老小鼠研究益生汤对小鼠心肌线粒体超氧化物歧化酶(SOD)活性,丙二醛(MDA)含量及肝组织中谷胱甘肽过氧化物酶(GSH-Px)活性的影响.结果:益生汤清除DPPH自由基、·OH自由基和O2-自由基的IC50值分别为0.547 0、0.209 1和0.300 9 mg·ml-1;总还原力的IC50为0.506 2 mg·ml-1;益生汤可明显抑制亚油酸的氧化.体内研究显示,与模型组相比,益生汤可剂量依赖性提高小鼠心肌线粒体中SOD的活性(P＜0.01)和肝组织GSH-Px活性(P＜0.01),降低心肌线粒体中MDA的含量(P＜0.01).结论:益生汤有一定的抗氧化功能,具有良好的开发前景.     

Anti-inflammatory,Antinociceptive, and Neuropharmacological Activities of Clerodendron viscosum.

Abstract

The crude methanol extract of Clerodendron viscosum. Vent. (Verbenaceae) leaves was evaluated for its anti-inflammatory, antinociceptive, and neuropharmacological activities. When given orally to rats at doses of 200 and 400 mg/kg of body weight, the extract showed a significant (p < 0.001) anti-inflammatory activity against carrageenan-induced rat paw edema comparable with the standard drug phenylbutazone at the dose of 100 mg/kg of body weight. It also produced a significant writhing inhibition in acetic acid–induced writhing in mice at the oral dose of 250 and 500 mg/kg of body weight (p < 0.001), which was comparable with the standard drug diclofenac sodium at the dose of 25 mg/kg of body weight. Moreover, when given intraperitoneally to albino mice, it potentiated the pentobarbital-induced sleeping time (p < 0.001), decreased the open field score in open field test (p < 0.001), decreased the number of holes crossed from one chamber to the other in the hole-cross test (p < 0.001), and decreased the head dip responses in the hole-board test (p < 0.001) at the dose of 250 and 500 mg/kg of body weight. The overall results tend to suggest the anti-inflammatory, antinociceptive, and central nervous system depressant activities of the crude methanol extract of Clerodendron viscosum..     

黄花蒿幼嫩叶的化学成分

目的研究黄花蒿(Artemisia annuaL.)全草的化学成分。方法用硅胶、聚酰胺和SephadexLH-20柱色谱分离化合物,根据理化性质和波谱数据鉴定其结构。结果从黄花蒿全草的乙醇提取物中分离得到13个化合物,分别鉴定为:青蒿素(artemisinin,1)、青蒿乙素(arteannuin B,2)、3α-羟基-1-去氧青蒿素(3α-hydroxy-1-deoxyartemisinin,3)、青蒿酸(artemisinic acid,4)、artemetin(5)、猫眼草黄素(chrysosplenetin,6)、quercetagetin-3,7,3′,4′-tetramethyl ether(7)、猫眼草酚(chrysosplenol D,8)、水杨酸(salicylic acid,9)、domesticoside(10)、东莨菪苷(scopolin,11)、β-谷甾醇(β-sitosterol,12)、胡萝卜苷(daucostrol,13)。结论化合物1-4为同一类化合物,属倍半萜类,化合物5-8为具有多甲氧基取代的黄酮类化合物,化合物10为首次从菊科植物中分离得到。     

Toxic essential oils. Part II: Chemical,toxicological, pharmacological and microbiological profiles of Artemisia annua L. volatiles

Botanical drugs based on Artemisia annua L. (Asteraceae) are important in the treatment of malaria. Alongside with artemisinin, this aromatic species produces high and variable amounts of other chemicals that have mostly unknown biological/pharmacological activities. Herein, we have studied the toxicological/pharmacological profile of volatile constituents of a Serbian population of A. annua. Fifty-eight components were identified, among them, artemisia ketone (35.7%), α-pinene (16.5%) and 1,8-cineole (5.5%) were the most abundant ones. Significant variability of A. annua volatile profile was confirmed by means of agglomerative hierarchical cluster analysis indicating the existence of several different A. annua chemotypes. In an attempt to connect the chemical profile of A. annua oil with its biological/toxicological effects, we have evaluated in vivo and/or in vitro toxicity (including hepato- and nephrotoxicity/protection), antinociceptive, antioxidant (DPPH, ABTS and superoxide radical scavenging activity assays), enzyme inhibiting (protein kinase A and α-amylase) and antimicrobial potential of A. annua oil and of its constituents. Our results revealed that the beneficial properties of A. annua botanical drugs are not limited only to their antimalarial properties. Taking into account its relatively low toxicity, the usage of A. annua volatiles (at least of the herein studied population) does not represent a health risk.