正天丸中钩藤的薄层色谱鉴别方法改进

目的：对正天丸原质量标准中钩藤的薄层色谱鉴别项下的供试液制备及层析条件进行改进，使检验结果更易判断。方法：对原标准中钩藤的薄层色谱鉴别项下的提取方法和层析及显色条件进行改进。经过酸碱交替提取，很大程度上排除了中性及酸性成分的干扰，展开剂改用碱性，使钩藤中的生物碱容易游离出来，选用生物碱显色剂碘化铋钾，使鉴定效果更加灵敏、稳定，阴性对照无干扰，斑点更清晰易分开。结果：操作简便，色谱斑点清晰、无干扰。结论：改进法更有利于检验结果的判定。     

Application of radioligand-receptor binding assays in the search for the active principles of the traditional Chinese medicine ‘Gouteng’

Uncaria rhynchophylla and related species (i.e. Gouteng of the Pharmacopoeia of the People's Republic of China) have antihypertensive, sedative and anticonvulsant activities. A methanol extract of ‘Gouteng’ (U. rhynchopylla) hooks and stems was assessed for its ability to inhibit the binding of radioligands to 13 different receptors in this study. The extract inhibited ligand-receptor binding by more than 60% to α₂-adrenoceptors, dopamine 1, 5-HT1A, opiate, GABA_A and GABA_B receptors. Bioassay-guided fractionation resulted in the isolation of ursolic acid (muscarinic and sulphonylureas activities), husutine (α₂- and β-adrenoceptor, 5-HT1A and 2, opiate and sulphonylureas activities) and epiallocorynanthine (β-adrenoceptor, 5-HT1A and 2, and opiate receptor activities). These findings are discussed in relation to the reported pharmacological properties, clinical uses and differing alkaloid profiles of those Uncaria species which constitute ‘Gouteng’ of the Pharmacopoeia of the People's Republic of China. © 1997 John Wiley & Sons, Ltd.     

正交实验法优选天麻钩藤饮的提取工艺

目的：优选天麻钩藤饮的提取工艺。方法：以天麻素含量为指标，采用正交设计，利用高效液相色谱法来考察煎煮时间、加水倍数、煎煮次数对天麻素提取效果的影响。结果：煎煮时间和煎煮次数对检测指标均有显著影响，加水倍数为不显著因素。结论：以10倍量的水煎煮3次，每次45min为最佳提取工艺。     

碳纤维微电极实时在线研究钩藤对小鼠脑内5-羟色胺的影响

目的 建立碳纤维微电极实时在线测定小鼠脑内海马区5-羟色胺含量的方法.方法 制备火焰刻蚀的碳纤维微电极,利用循环伏安法,建立生理pH条件下5-羟色胺与氧化峰电流的线性关系.在线测定钩藤给药小鼠脑内海马区5-羟色胺随时间的变化,同时与空白组和安定对照组小鼠进行比较.结果 5-羟色胺在碳纤维微电极上的氧化峰电位为0.40 V,峰电流与5-羟色胺在1.0× 10-8～1.0× 10-5 mol/L范围内呈良好的线性关系,检测限为1.0× 10-8 mol/L.钩藤给药小鼠脑内海马区5-羟色胺在给药1h后含量最高,随后呈逐渐降低的趋势.结论 碳纤维微电极制作简单,稳定性和重现性良好,火焰刻蚀后灵敏度大大提高,干扰少.该法用于活体实时在线测定,结果与药理实验具有一致性.     

Effect of Alkaloid-Free and Alkaloid-Rich preparations from Uncaria tomentosa bark on mitotic activity and chromosome morphology evaluated by Allium Test

Ethnopharmacological revelance

Uncaria tomentosa (Willd.) DC. is the most popular Peruvian plant, used in folk medicine for different purposes. It contains thousands of active compounds with great content of alkaloids.

Aim of study

Two different fractions of Alkaloid-Rich and Alkaloid-Free were researched on chromosome morphology, mitotic activity and phases indexes.

Materials and methods

Cells of Allium Test (meristematic cells of root tips) were incubated up to 24 h in different concentrations of Alkaloid-Free and Alkaloid-Rich fraction obtained from Uncaria tomentosa bark followed by 48 h of postincubation in water. The chromosome morphology was analyzed and the content of mitotic and phase indexes were done. Individual compounds, oxindole alkaloids, phenolic compounds and sugars were determined.

Results

In Alkaloid-Rich and Alkaloid-Free fractions (different in chemical composition) we observed condensation and contraction of chromosomes (more in Alkaloid-Rich) with retardation and/or inhibition of mitoses and changed mitotic phases. Postincubation reversed results in the highest concentration which was lethal (in mostly Alkaloid-Rich fraction).

Conclusions

Our studies indicate that different action can depend on different groups of active compounds in a preparation either containing alkaloids or not. Other fraction analysis may be useful in the future.     

大叶钩藤叶子化学成分研究

目的：对茜草科钩藤属植物大叶钩藤（Uncaria macrophylla Wall.）的叶子进行化学成分研究。方法：用甲醇提取及柱色谱等方法进行分离,薄层色谱跟踪检测,波谱法鉴定结构。结果：从该植物叶子中共分离鉴定出6个化合物,分别是柯诺辛碱（1）、、异钩藤碱（2）、钩藤碱（3）、熊果酸（4）、表儿茶素（5）、山奈酚（6）。结论：该实验首次报道了大叶钩藤叶子的非挥发性化学成分。     

滇钩藤化学成份研究

目的　研究滇钩藤(Uncaria yunnanensis Hsia.C.C)根的化学成分。方法　运用各种色谱技术进行分离,用IR,MS,¹HNMR,¹³CNMR,2DNMR等光谱技术鉴定化合物。结果　得到7个化合物,分别鉴定为:3β,6β,19α-三羟基乌苏酸(I),23,-去甲基24烯-3β,6β,19α-乌苏酸(II),6β,19α-二羟基-乌苏酸-3-酮(III),齐墩果酸(IV),5,7,3′,4′-四羟基-黄烷-3-醇(V),β-育亨宾(VI)和滇钩藤碱I(VII)。结论　VII为新化合物,其余化合物均为首次从该植物中分得。     

钩藤总碱预处理对海马神经元急性缺氧的保护作用

目的:观察钩藤总碱预处理对急性缺氧后海马神经元电压门控性Na⁺电流的影响。方法:原代培养大鼠海马神经元,分为常氧、急性缺氧和钩藤总碱预处理进行实验。采用膜片钳全细胞记录技术,检测海马神经元钠电流(I_Na)的变化。结果:急性缺氧后,细胞I_Na幅度明显降低,I_Na阈值右移;而经钩藤总碱预处理的海马神经元I_Na的降低幅度明显减轻。结论:钩藤总碱预处理提高了海马神经元对急性缺氧的耐受性,对神经元有显著保护作用。     

天麻钩藤饮研究进展

天麻钩藤饮现广泛应用于心血管系统及神经系统等多种疾病,对其进行的研究也从药理、药效、质量控制等方面逐渐深入,研究取得了一定进展,但仍存在较多问题。(1)在临床应用方面,运用的方法学质量普遍较低,对天麻钩藤饮治疗疾病的疗效评价多单纯运用主观症状的改善来衡量,客观指标过少,影响了实验的可靠性和说服力。(2)在药理研究方面,天麻钩藤饮药理作用复杂,目前对其进行的研究以血清药理学方法为主,但试验指标相对滞后,重复性研究较多,突破性进展较少。此外,中药复方药理研究不能脱离中医药理论的指导作用,不能忽略病与证的结合问题,故规范动物试验模型是试验成功的前提,同时应密切结合其病理变化寻找研究方向。(3)天麻钩藤饮复方的药效物质基础方面目前还未见相关文献报道,由于中药复方的药效物质基础能够阐明其药用功效、作用机制,是其安全性和质量控制的保障,故从整体入手研究天麻钩藤饮的药效物质基础将是其必不可少的研究方向。     

非水毛细管电泳法测定钩藤中的钩藤碱和异钩藤碱

目的 建市同时测定钩藤中钩藤碱、异钩藤碱含量的方法.方法 采用非水毛细管电泳法,运行缓冲液为0.05 mol·L~(-1)的醋酸铵甲醇溶液(冰醋酸凋pH6.4),操作电压25 kV,温度为20℃,检测波长为239 nm.结果 钩藤碱0.014～0.570mg·mL~(-1)与峰面积呈良好的线性关系(r=0.9997),加样同收率为98.8%,RSD=2.34%;异钩藤碱0.008～0.194 mg·mL~(-1)与峰面积呈良好的线性关系(r=0.9996),加样回收率为96.3%,RSD=3.06%.钩藤碱及异钩藤碱可良好地分离.结论 所建方法可快速有效地分离钩藤碱及异钩藤碱并测定植物中钩藤碱的含量.