中药大黄的綜合研究 Ⅱ.蒽醌衍生物的紙上层析

本文研究了中药大黄中蒽醌衍生物纸上层析的扩展剂体系和其他条件.实验试用了33种扩展剂体系,我们选择了8种效果较好者,其中以四氯化碳-苯-水体系为最好.以此改进的纸上层析法进行了不同品种大黄中蒽醌衍生物含量的测定,实验结果表明,不同品种及同品种不同部位的大黄中各种蒽醌衍生物的含量差异很大.蒽醌衍生物总量及致泻与抗菌有效成分,都以西宁大黄为最高.     

市售三黄片中游离和结合蒽醌含量的比较及聚类分析

目的：比较市售不同厂家三黄片中游离蒽醌和结合蒽醌芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚及其苷的含量,并进行聚类分析。方法：采用HPLC法,色谱柱为Eclipse plus C₁₈色谱柱,流动相为甲醇-0.1%磷酸溶液,梯度洗脱,流速为0.8 mL·min^-1,检测波长为254 nm,柱温为25℃,进样量为20 μL。结果：测得10个不同厂家三黄片中游离总蒽醌含量分别为6.272,9.143,3.950,6.718,7.214,5.411,8.501,5.411,11.725,8.785 mg·g^-1,结合总蒽醌含量分别为3.781,0.700,6.060,4.202,5.560,8.369,6.036,5.621,1.624,3.361 mg·g^-1。聚类分析结果表明,类别的划分突出特性,反映蒽醌含量具有空间分布特征,不同厂家最高和最低的含量存在很大差异。结论：10个厂家三黄片中大黄总蒽醌含量差别大,特别是作为三黄片发挥泻下药效作用的结合蒽醌含量也相差悬殊。     

黄芩对大黄蒽醌在提取精制过程中转化的影响

目的探讨黄芩对大黄中5种蒽醌类成分在提取精制过程中相互转化的影响。方法 HPLC测定药材和提取物中5种成分的含量,比较药材和提取物中芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的相对比例;再分别用5种蒽醌的对照品加入和不加黄芩药材提取液模拟提取精制过程,考察5种蒽醌成分之间的转化情况。结果大黄药材和提取物中5种成分的相对比例有明显变化,大黄酸在提取物中的比例增高;同时采用对照品模拟提取精制过程时各个成分并无转化,而加入黄芩药材提取液后,大黄酸经过提取精制处理后能部分转化为大黄素,其余4种成分之间无转化。结论黄芩和大黄药材配伍提取可以使大黄蒽醌类成分发生转化。     

中药大黄的综合研究 Ⅰ.大黄中蒽醌衍生物抗菌效价的研究

中药大黄在祖国医学經驗中不但用以緩下、健胃及利胆,并且亦作为清毒消炎之用。关于大黄水煎剂抗菌作用的实驗研究,文献已有报告;但关于其抗菌作用有效成分的研究报告很少。据文献,Costa Rica地方民間曾用Cassia reticulata叶的浸出物口服以治疗淋病。1947年,Robbias,Kavanagh和Thayer等曾从此叶分离出一种Cassicacid,发現其对金黄色葡萄球菌、枯草杆菌、草分枝杆菌、淋病双球菌、蕈状菌及包皮垢菌等有显著抑制作用,其抑菌最低浓度为4—32微克/毫升;但对大腸杆菌、肺炎杆菌及綠脓     

高效液相色谱法测定狗体内大黄游离蒽醌的含量

目的 :建立狗体内大黄游离蒽醌的含量测定方法。方法 :采用HypersilBDSC18 色谱柱 (4 6mm×250mm) ,甲醇 -0 5 %冰醋酸 (80∶20)为流动相 ,流速为1 0ml/min ,检测波长为430nm。样品以甲醇沉淀蛋白并提取 ,常温下氮气吹干 ,复溶进样分析。结果 :芦荟大黄素在0 15～6 0μg/ml(r=0 9996)、大黄酸在0 14～5 6μg/ml(r=0 9982)、大黄素在0 13～5 2μg/ml(r=0 9990)、大黄酚在0 2～8 0μg/ml(r=0 9991)、大黄素甲醚在0 1～4 0μg/ml(r=0 9994)范围内线性关系良好。结论 :该方法为大黄制剂中游离蒽醌成分的体内分析及其生物利用度研究提供了方法依据。     

生熟大黄总提取物灌胃给药后游离蒽醌在SD大鼠组织中的分布差异研究

大黄应用范围广、疗效确切,然而近年来国内外实验报道大黄中蒽醌成分具有肝、肾细胞毒性。本文对比研究了生熟大黄总提取物灌胃给药后游离蒽醌在正常SD大鼠组织中的分布。肝、脾和肾组织中游离蒽醌类成分含量采用UPLC-MS/MS法测定。生熟大黄总提物(相当于生药量14.69 g.kg-1)灌胃给药,连续12周。结果表明,游离蒽醌在生大黄组大鼠组织中的分布浓度高于熟大黄组;大黄游离蒽醌在同一组织中的分布浓度顺序大致为大黄酸>大黄素>芦荟大黄素;大黄酸在肝、脾和肾组织中的分布浓度明显高于芦荟大黄素和大黄素,且大黄酸在生大黄组的组织分布浓度明显高于熟大黄组;停药恢复4周后,大黄游离蒽醌在组织中检测不到。结果提示生大黄的组织毒性可能高于熟大黄;大黄酸可能是大黄主要的毒性物质之一;大黄游离蒽醌在组织中可能无蓄积毒性。炮制过程不仅影响了有效成分的含量还可能影响其成分的组成,通过影响吸收和作用过程中成分之间的相互作用,致使生大黄组动物组织中游离蒽醌的分布浓度高于熟大黄组,这与传统中医药理论炮制减毒的思想认识基本一致。     

碱提法提取大黄游离蒽醌的研究

目的优选碱提法提取大黄游离蒽醌的最佳提取工艺.方法比较浸渍法、加热回流法、超声提取法3种提取方式,并采用L9(34)正交试验,以大黄游离蒽醌的含量为指标,对影响大黄游离蒽醌碱法提取工艺的因素水平进行了研究.结果碱提法提取大黄游离蒽醌的最佳提取工艺为:大黄粉末用15倍量2%的Na2CO3溶液提取两次,每次30min,提取温度为20℃.结论碱提法提取大黄游离蒽醌的优选工艺稳定可行.     

Chrysophanol-8-O-glucoside, an anthraquinone derivative in rhubarb, has antiplatelet and anticoagulant activities

Rhubarb is a widely used traditional medicine and has been reported to elicit a number of biological effects including anti-inflammatory and antiplatelet effects. In the present study, we investigated the effects of anthraquinone derivatives isolated from rhubarb on platelet activity. Of four anthraquinone derivatives isolated from rhubarb examined, chrysophanol-8-O-glucoside (CP-8-O-glc) was found to have the most potent inhibitory effect on collagen- and thrombin-induced platelet aggregation. CP-8-O-glc-treated mice showed significantly prolonged bleeding times. Furthermore, CP-8-O-glc was found to have a significant inhibitory effect on rat platelet aggregation ex vivo and on thromboxane A(2) formation in vitro. In coagulation tests, CP-8-O-glc did not alter prothrombin time, and it prolonged the activated partial thromboplastin time. However, CP-8-O-glc only inhibited platelet phosphatidylserine exposure, but not exert direct inhibition on intrinsic factors. This study demonstrates the antiplatelet and anticoagulant effects of CP-8-O-glc and suggests that this compound might be of therapeutic benefit for the prevention of platelet-related cardiovascular diseases.     

Simultaneous determination of eight active components in Chinese medicine 'YIQING' capsule using high-performance liquid chromatography

An effective, accurate and reliable method for the simultaneous separation and determination of eight active components (berberine, aloe-emodin, rhein, emodin, chrysophanol, baicalin, baicalein and wogonin) in Chinese medicine 'YIQING' capsule was developed using reverse phase high-performance liquid chromatography (RP-HPLC) coupled with diode array detection. The chromatographic separation was performed on a Lichrospher C18 column (250 mm x 4.6 mm i.d. with 5.0 microm particle size) with a simple linear gradient elution programme. Due to the different UV characteristic of these components, three detection wavelengths were utilized for the quantitative analysis (UV wavelength 254 nm for anthraquinone derivatives, 278 nm for flavones compounds, and 345 nm for protoberberine alkaloids, respectively). Excellent linear behaviors over the investigated concentration ranges were observed with the values of R2 higher than 0.99 for all the analytes. The recoveries, measured at three concentration levels, varied from 94.9% to 105.3%. The validated method was successfully applied to the simultaneously determination of these active components in 'YIQING' capsules from different production batches.     

清肠饮中大黄蒽醌含量的比色分析方法研究

目的:建立一种测定清肠饮中大黄蒽醌含量的比色分析方法.方法:采用5%氢氧化钠 2%氢氧化铵混合碱液和0.5%醋酸镁 甲醇液两种比色法对样品的含量进行比较研究.结果:醋酸镁 甲醇比色法灵敏、简便、准确且稳定性好.结论:醋酸镁 甲醇比色法可作为测定大黄中蒽醌类含量的常规分析方法.     

新型骨靶向抗肿瘤大黄酚衍生物的合成

目的: 利用大黄蒽醌类化合物的抗肿瘤作用与趋骨性,将大黄酚抗肿瘤药5-氟脲嘧啶及其衍生物连接,合成系列新型骨靶向抗肿瘤衍生物.方法: 合成大黄酚衍生物,MTT法测定其对肿瘤细胞增殖的抑制作用,用羟基磷灰石吸附试验评价该类药物的体外骨亲和性.结果: 合成了21个大黄酚衍生物均为新化合物.实验显示所有化合物均有不同程度的骨亲和性,大部分化合物的亲和性高于阳性对照药四环素.结论: 大黄酚衍生物具有良好的骨亲和性.     

药用大黄中蒽醌和非蒽醌类成分的分离与鉴定

目的对药用大黄中蒽醌及非蒽醌类成分进行深入研究。方法药用大黄80%(φ)乙醇溶液提取物经硅胶柱、ODS柱、MCI gel CHP20、SephadexLH-20凝胶柱色谱及HPLC色谱分离,纯化,根据理化性质和波谱数据进行结构鉴定。结果分离得到6个蒽醌类和5个非蒽醌类成分,经波谱数据解析,分别鉴定为香兰基丙酮(vanillylacetone,1)、大黄素(emodin,2)、芦荟大黄素(aloeemodin,3)、大黄素甲醚-8-O-β-D-葡萄糖苷(physcion-8-O-β-D-glucopyranoside,4)、大黄酚-8-O-β-D-葡萄糖苷(chrysophanol-8-O-β-D-glucopyranoside,5)、1-甲基-8-羟基-9,10-蒽醌-3-O-β-D-(6'-O-桂皮酰基)吡喃葡萄糖苷(1-methyl-8-hydroxyl-9,10-anthraquinone-3-O-β-D-(6'-O-cinnamoyl)glucopyrano-side,6)、芦荟大黄素-3-(羟甲基)-O-β-D-葡萄糖苷(aloe emodin-3-(hydroxymethyl)-O-β-D-glucopy-ranoside,7)、(+)儿茶素(catechin,8)、表儿茶素没食子酸酯(epicatechin-3-O-gallate,9)、儿茶素-3-O-β-D-葡萄糖苷(catechin-3-O-β-D-glucopyranoside,10)、儿茶素-8-β-D-葡萄糖苷(catechin-8-β-D-glucopyranoside,11)。结论化合物1,6,9-11为首次从该植物中分离。     

Lipoxygenase inhibitory constituents from rhubarb

Phytochemical study on the ethanol extract of rhubarb led to the isolation of fifteen compounds, including five anthraquinones: chrysophanol (1), physcion (2), emodin (7), chrysophanol-8-O-beta-D: -glucopyranoside (9) and emodin-8-O-beta-D: -glucopyranoside (15), and ten stilbenes: desoxyrhaponticin (3), rhaponticin (4), resveratrol (5), desoxyrhapotigenin (6), rhapontigenin (8), piceatannol-3'-O-beta-D: -glucopyranoside (10), piceid (11), epsilon-viniferin (12), ampelopsin B (13) and isorhaponticin (14). Their structures were identified by comparing the physicochemical data with those of published papers. Among the isolated compounds, stilbene derivatives (3-6, 8 and 10-14) showed remarkable inhibitory effect on lipoxygenase with IC(50) values ranging from 6.7 to 74.1 microM. The inhibition kinetics analyzed by Lineweaver-Burk plots found that they were competitive inhibitors with the linoleic acid at the active site of lipoxygenase. In addition, stilbenes exhibited significantly free radical scavenging activity against ABTS(.+) with trolox equivalent activity capacity (TEAC) values ranging from 1.16 to 4.64. Whereas, anthraquinone derivatives (1-2, 7, 9 and 15) neither inhibited lipoxygenase nor scavenged free radical ABTS(.+). These results indicated that stilbene derivatives were considerate to be mainly lipoxygenase inhibitor and free radical scavenger constituents of rhubarb.     

Comparison of phenolic compounds of rhubarbs in the section deserticola with Rheum palmatum by HPLC-DAD-ESI-MSn

A sophisticated LC-ESI-MS (n) method was utilized for the analysis of the phenolic compounds in five unofficial RHEUM species for the first time. These species belong to the section deserticola Maxim. of the Polygonaceae family, including Rheum racemiferum Maxim., R. nanum Siev. ex Pall., R. delavayi Franch., R. sublanceolatum C. Y. Cheng et T. C. Kao and R. pumilum Maxim. A total of 101 phenolic compounds, including sennosides, anthraquinones, stilbenes, glucose gallates, naphthalenes, procyanidins and chromones were identified or tentatively characterized based on their UV and mass spectral data from the methanol extracts. These chemical constituents were compared with those of an official species ( R. palmatum l.). The results showed that most sennosides and procyanidin derivatives were only present in R. palmatum rather than in the unofficial species; stilbenes and chromones were only found in the sect. deserticola species; while anthraquinones, glucose gallates and naphthalenes could be detected in all studied species. For anthraquinone glycosides, only emodin O-glucosides were detected in the sect. deserticola species. Moreover, remarkable difference was observed among unofficial species. These results may be useful for the quality control of rhubarb in order to guarantee its clinical effects, and may also be helpful for the further clarification of the taxonomic relationship of the plants in the sect. deserticola.     

高效液相色谱法测定何首乌和夜交藤中蒽醌类成分的含量

建立了何首乌中的主要有效成分大黄素、大黄素甲醚、大黄酚及大黄酸等蒽醌类的反相高效液相色谱法。以ＴＳＤ－ｇｅｌ８０Ｔｓ为固定相，甲醇－水－磷酸（７２０：２８０：１）为流动相。样品用７０％甲醇提取浓缩后用氯仿萃取，分离出游离型和结合型蒽醌的定量用试液。本法精确、可靠。用此方法测定了不同产地的９种何首乌和４种夜交藤样品。     

5个蒽醌三三配伍治疗脑缺血大鼠的药代动力学研究

目的:建立LC-MS方法测定大鼠血浆中蒽醌成分含量,探讨5个大黄蒽醌成分(芦荟大黄素、大黄素、大黄酸、大黄酚、大黄素甲醚)三三配伍时在脑缺血大鼠模型中的药代动力学特征。方法:MCAO法制备大鼠局灶性脑缺血模型。模型大鼠随机分成11组,空白组1组,给药组10组。给药组将5个大黄蒽醌三三结合,给药剂量:芦荟大黄素(A)13.05 mg·kg^-1,大黄素(B)34.65 mg·kg^-1,大黄酸(C)17.25mg·kg^-1,大黄酚(D)39.45 mg·kg^-1,大黄素甲醚(E)26.4 mg·kg^-1。血浆样品经甲醇沉淀蛋白后,XBridge^TMC₁₈色谱柱分析,高分辨质谱检测。流动相为甲醇-3 mmol·L^-1乙酸铵,梯度洗脱。采用内标法测血药浓度,Kinetica 5.0药动学软件以非房室模型计算药代动力学参数。结果:当与不同的2个大黄蒽醌联用时,芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚的药代动力学参数均存在差异。结论:本研究为这5个大黄蒽醌的作用机制研究提供理论基础,为其临床用药提供了实验参考。     

决明子发酵前后5种蒽醌类成分变化研究

目的:探讨固态发酵对决明子中5种蒽醌类成分含量的影响。方法:采用高效液相色谱法测定决明子发酵前后芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的含量并进行对比研究。色谱柱为C18-ODS反相柱(250mm×4.6mm,5μm),流动相为甲醇-0.1%磷酸水溶液(梯度洗脱),流速为1mL.min-1,检测波长为280nm。结果:决明子经过发酵后,游离型蒽醌含量呈上升趋势。以未发酵成分含量为100%计算,5种成分变化分别为104.6%、102.2%、93.4%、215.6%、134.8%,其中芦荟大黄素、大黄酸和大黄素变化不明显,大黄酚与大黄素甲醚分别增加至2.15倍和1.34倍,可见发酵增加了蒽醌苷元的含量。结论:决明子通过发酵,有利于增加活性成分蒽醌苷元的含量。     

Photoreactivity of anthraquinones for the analysis of ginsenosides using photoreduction fluorescence detection-HPLC

The photoreactivity of twelve anthraquinone derivatives was examined to evaluate its usefulness as a photo-reagent for the analysis of ginsenosides using photoreduction fluorescence (PRF) detection method. Among the tested compounds, 2-tert-butylanthraquinone (TBAQ), 2-chloroanthraquinone (CAQ) and anthraquinone (AQ) showed good characteristics as photoreagents. The detection limits of ginsenoside Rg₁ by PRF-HPLC method using TBAQ, CAQ or AQ as a photo-reagent were found to be ca. 35 ng, 50 ng and 50 ng, respectively.     

HPLC法测定微米大黄炭中没食子酸的含量

目的：建立HPLC法测定微米大黄炭中没食子酸含量。方法：采用Fortis universil C18色谱柱（250 mm×4.6 mm,5μm）;流动相为甲醇：0.1%磷酸溶液（12：88）;检测波长：273 nm;流速：0.9 ml·min-1。结果：没食子酸回归方程为Y=31.93X＋2.61（r=0.999 9）,其浓度线性范围6.666.0μg·ml-1。平均回收率为101.35%,RSD=1.16%（n=6）。结论：测定方法简便、准确、重复性好,可用于微米大黄炭中没食子酸的含量测定。