瘤果槲寄生及其3种寄主齐墩果酸和熊果酸的含量研究

目的:考察瘤果槲寄生及其3种寄主植物齐墩果酸和熊果酸含量的关系。方法:采用HPLC法对瘤果槲寄生及其寄主植物齐墩果酸、熊果酸含量进行测定,齐墩果酸、熊果酸采用乙醇超声提取30 min,用Agilent Eclipse XDB-C18柱,甲醇-乙腈-0.05%醋酸铵溶液(12∶67∶21)为流动相,流速1.0mL·min-1,检测波长210 nm。结果:齐墩果酸和熊果酸的线性范围分别为3.91～313 mg·L-1(r=0.999 8)和8.15～652 mg·L-1(r=0.999 9),与峰面积呈良好线性关系,平均加样回收率分别为99.27%和99.75%。结论:瘤果槲寄生含齐墩果酸,不含熊果酸。瘤果槲寄生与其寄主齐墩果酸含量没有直接关系,为其自身固有。     

反相高效液相色谱法测定槲寄生中齐墩果酸的含量

［摘要］目的采用反相高效液相色谱（RP HPLC）法测定槲寄生中齐墩果酸的含量。方法采用Hypersil ODS2柱（250 mm ×4.6 mm，5 μm），流动相为甲醇：1.0%冰醋酸水溶液（94：6），检测波长为210 nm ，流速为0.5 mL•min 1。结果齐墩果酸分别在1.002～5.010 μg，范围线性关系良好(r＝0.998 3)；齐墩果酸的平均回收率为99.98%(RSD＝2.81%)。结论该方法简便，准确、重复性好，适用于槲寄生药材中齐墩果酸的质量控制。     

广寄生和瘤果槲寄生及其寄主植物中齐墩果酸、熊果酸含量的相关性研究

目的:考察寄主及其寄生植物齐墩果酸和熊果酸含量的关系.方法:采用HPLC法对寄主及其寄生齐墩果酸、熊果酸含量进行测定,样品采用乙醇超声提取30 min,用Agilent Eclipse XDB-C18柱(250 mm×4.6 mm,5 μm),甲醇-乙腈-0.05%醋酸铵溶液(12:67:21)为流动相,流速为1.0 ml·min-1,检测波长为210 nm.结果:齐墩果酸和熊果酸的线性范围分别为2.61～209.00 mg·L-1(r=0.999 8)和7.85～628.00 mg·L-1(r=0.999 9),浓度与峰面积呈良好线性关系,平均加样回收率分别为99.27%和99.75%.结论:广寄生不含齐墩果酸和熊果酸,瘤果槲寄生中含有齐墩果酸,不含熊果酸.     

球形红细菌生物转化槲寄生中总黄酮类化合物的测定

目的建立槲寄生及球形红细菌转化槲寄生培养液中总黄酮类化合物的含量测定方法,并对各个样品的含量进行比较研究。方法采用紫外分光光度法,以芦丁为对照品,以亚硝酸钠一硝酸铝溶液为显色系统．512nm波长处测定总黄酮类化合物的含量。结果芦丁在8-48μg／ml的范围内线性关系良好,回归方程为l,=10．627X-0．0544,r=O．9979。槲寄生75％的乙醇提取物（样品3）、水提取物（样品6）经过球形红细菌转化总黄酮类化合物的含量分别增加129％、65％。结论经过球形红细菌转化可以增加槲寄生提取液中总黄酮类化合物的含量。     

高效液相色谱法测定不同产地白花蛇舌草中齐墩果酸和熊果酸的含量

目的分析不同产地白花蛇舌草中齐墩果酸和熊果酸含量,为药材及其制剂的质量控制提供参考。方法采用高效液相色谱法(HPLC)测定。色谱柱AgilentC18柱,流动相为甲醇-水-冰醋酸-三乙胺(91.5∶8.5∶0.04∶0.1),检测波长210nm,流速0.80ml/min,柱温20℃。结果齐墩果酸和熊果酸分别在20.4～326.4μg/ml(r=0.9991)和39.2～627.2μg/ml(r=0.9994)范围内线性关系良好;齐墩果酸和熊果酸的平均回收率在98.0%～99.0%,相对标准差(RSD)分别为0.87%和1.22%。结论本方法准确、简便、重现性好,可同时测定白花蛇舌草中齐墩果酸和熊果酸的含量,能够应用于药材及其制剂的质量控制。     

柱前衍生化HPLC-UV测定牛膝中齐墩果酸的含量

目的建立牛膝中齐墩果酸的柱前衍生化高效液相色谱测定法。方法以对硝基苯甲酰氯为衍生化试剂,对齐墩果酸分子中的羟基进行苯甲酰化,以乙腈-0.2%磷酸水溶液（95∶5）为流动相,在254nm波长处检测。结果齐墩果酸衍生化产物线性回归方程为：Y=85197X＋22550（r=0.9998）,表明齐墩果酸衍生产物在2.0～6.0μg内线性关系良好。结论所建立的柱前衍生化HPLC-UV可用于牛膝中齐墩果酸的含量测定,齐墩果酸与对硝基苯甲酰氯反应,衍生物为对硝基苯甲酸齐墩果酸酯,紫外检测波长由210nm红移至254nm,与以往方法比较,大幅改善了测定效果,提高了检测灵敏度。该方法准确,灵敏度高,为牛膝药材的质量评价提供了参考依据。     

高效液相色谱法测定山楂药材中齐墩果酸和熊果酸的含量

目的应用高效液相色谱法（HPLC）测定山楂药材中齐墩果酸和熊果酸的含量,并以齐墩果酸和熊果酸的含量为标准建立山楂的质量标准。方法色谱条件HypersilODSC18（150mm×4.6mm,5μm）;流动相为甲醇—水（88：12）;检测波长210nm,流速0.8ml/min,柱温25℃。结果齐墩果酸的线性范围为0.172～1.610μg,r=0.9993,回收率为97.98%（RSD=1.39%）;熊果酸的线性范围为0.473～4.117μg,r=0.9995,回收率为97.21%（RSD=1.27%）。结论该方法简便、可靠、准确,可用于山楂药材中齐墩果酸和熊果酸的含量测定和山楂的质量控制。     

HPLC法测定齐墩果酸胶囊中齐墩果酸的含量

建立高效液相色谱法测定齐墩果酸胶囊的含量.采用HYPERSIL C18色谱柱;流动相:甲醇-0.1%磷酸溶液(90:10);流速:1.0mL·min-1;检测波长:210nm.齐墩果酸在1.018～10.18цg范围内线性关系良好(r=1.0000);齐墩果酸平均回收率99.6%,RSD=0.8%.该方法灵敏、准确,可用于齐墩果酸胶囊的含量测定.     

高效液相色谱法测定山楂药材中齐墩果酸和熊果酸的含量

目的 应用高效液相色谱法(HPLC)测定山楂药材中齐墩果酸和熊果酸的含量,并以齐墩果酸和熊果酸的含量为标准建立山楂的质量标准.方法 色谱条件Hypersil ODS C18(150mm×4.6mm,5μm);流动相为甲醇-水(88:12);检测波长210nm,流速0.8ml/min,柱温25℃.结果 齐墩果酸的线性范围为0.172～1.610μg,r＝0.9993,回收率为97.98%(RSD=1.39%);熊果酸的线性范围为0.473～4.117μg,r=0.9995,回收率为97.21%(RSD=1.27%).结论 该方法简便、可靠、准确,可用于山楂药材中齐墩果酸和熊果酸的含量测定和山楂的质量控制.     

不同产地对九节菖蒲中总皂苷、齐墩果酸及腺苷含量的影响

目的测定不同产地九节菖蒲中总皂苷、齐墩果酸及腺苷的含量。方法以齐墩果酸为对照品,采用紫外分光光度法于208 nm波长处对九节菖蒲中的总皂苷含量进行测定;采用高效液相色谱法对九节菖蒲中齐墩果酸的含量进行测定,色谱柱为Diamonsil C18柱(250 mm×4.6 mm,5μm),柱温为室温(25℃),流动相为乙腈-水(92∶8),流速为1 mL/min,检测波长为208 nm;采用高效液相色谱法对九节菖蒲中腺苷的含量进行测定,色谱柱为Diamonsil C18(250 mm×4.6 mm,5μm)柱,柱温为室温(25℃),流动相为乙腈-水(8∶92),流速为1 mL/min,检测波长为260 nm。结果不同产地九节菖蒲中总皂苷、齐墩果酸的含量差异较大,腺苷的含量差异较小。结论不同产地对九节菖蒲中总皂苷、齐墩果酸的含量有一定影响。     

紫外可见分光光度法测定苣荬菜根部总三萜皂苷的含量

目的用紫外可见分光光度法测定苣荬菜根部总三萜皂苷的含量。方法采用AB-8、D-101、D-4020、HPD-400和NKA-95种大孔树脂对苣荬菜根部总三萜皂苷进行纯化,以齐墩果酸为对照品,50g·L-1香草醛-冰醋酸、高氯酸为显色系统,在546nm波长处测定总三萜皂苷含量。结果在AB-8、D-101、D-4020、HPD-400和NKA-9 5种树脂中,D-101对苣荬菜根部总三萜皂苷纯化效果最好;齐墩果酸质量浓度在0.034 2⁰.106 5mg·mL-1范围内,吸光度与含量呈良好的线性关系,回归方程Y=8.298 2 X-0.084 9(r=0.999 6),平均加样回收率为95.3%,RSD=1.5%(n=5),苣荬菜根部总三萜皂苷含量为0.202%,RSD=2.6%。结论该方法操作简单,快速灵敏,准确可靠,对设备要求不高,适用于苣荬菜根部总三萜皂苷的含量测定。     

Box-Behnken设计-响应面法优化山茱萸叶总三萜提取工艺

目的:优化山茱萸叶总三萜提取工艺。方法:在山茱萸叶充分溶胀的基础上,采用加热回流法提取其总三萜。在单因素试验中考察乙醇体积分数、液料比、提取时间和提取次数对山茱萸叶总三萜含量的影响;以齐墩果酸为对照品,采用紫外-可见分光光度法测定总三萜含量。在单因素试验的基础上,固定提取次数为3次,以总三萜含量为响应值,以乙醇体积分数、液料比、提取时间为响应因素,采用Box-Behnken设计-响应面法优化山茱萸叶总三萜的提取工艺并进行验证。结果:山茱萸叶总三萜的最佳提取工艺为乙醇体积分数73%、液料比38∶1(mL/g)、提取时间60 min。3次验证试验结果显示,山茱萸叶总三萜的含量分别为6.92%、6.91%、6.84%,平均含量为6.89%(RSD=0.63%),与预测值(7.28%)的相对误差为5.36%。结论:优化的提取工艺稳定、可靠,可用于山茱萸叶总三萜的提取。     

Protective effects of betulin and betulinic acid against ethanol-induced cytotoxicity in HepG2 cells

Plant triterpenes, such as oleanolic acid and betulin were described as hepatoprotectants active against cytotoxicity of acetaminophen or cadmium. The aim of this paper is to compare the cytoprotective activity of betulin, betulinic acid and oleanolic acid against ethanol-induced cytotoxicity in HepG2 cells. The influence of three triterpenes on ethanol-induced production of superoxide anion and hydrogen peroxide was also examined. Among the examined triterpenes, betulin was the most active protectant of HepG2 cells against ethanol-induced cytotoxicity. Betulin and betulinic acid significantly decreased ethanol-induced production of superoxide anion. Oleanolic acid inhibited only ethanol- and phorbol ester-induced production of hydrogen peroxide. The results indicate that cytoprotective or antioxidative activity of triterpenes depends on their chemical structure.     

Antioxidant and antiatherogenic activities of pentacyclic triterpenic diols and acids

The present paper aimed to test the potential cardioprotective activity of four pentacyclic triterpenes, uvaol, erythrodiol, oleanolic acid and maslinic acid, widely distributed throughout the vegetable kingdom. For this purpose, their antioxidant and antithrombotic activities related to LDL particles have been in vitro evaluated. Results demonstrated that maslinic acid, uvaol and erythrodiol exert antiatherogenic effect while no effect was observed for oleanolic acid. Specifically, maslinic acid has shown the most potent dose-dependent antioxidant effect and did not have antithrombotic properties, whereas uvaol and erythrodiol exhibited both antioxidant and antithrombotic activities. In addition, antioxidant mechanisms of action were determined. While maslinic acid possesses dual activity acting as scavenger of free radicals and as copper chelator, uvaol is able to form a complex with copper and erythrodiol seems to behave as a retarder antioxidant. In conclusion, dietary triterpenes may exert a cardioprotective effect by different mechanisms of action related to antioxidant and antithrombotic activities.     

Effects of oleanolic acid and maslinic acid on hyperlipidemia

In the present study, we examined the therapeutic effects of pentacyclic triterpenes, oleanolic acid (OA) and maslinic acid (MA), on hyperlipidemia in a high‐cholesterol diet model. Hyperlipidemia was induced in male Sprague‐Dawley rats by feeding with a high‐cholesterol diet (HCD) for 30 days. MA and OA were supplemented (100 mg/kg body wt/day) during the last 15 days. The levels of serum total cholesterol (TC), triglyceride (TG), high‐density lipoprotein‐cholesterol (HDL‐C), and low‐density lipoprotein‐cholesterol (LDL‐C) increased in hyperlipidemic rats. An apparent increase in the expression of Acyl‐CoA cholesterol acyltransferase (ACAT) mRNA was seen in HCD‐fed rats. The activities of hepatic marker enzymes that serve as indices of cellular injury were also altered in HCD‐fed rats. Treatment with triterpenes modulated the abnormalities induced by hyperlipidimia. Lipid accumulation was decreased in histological findings. Elevated hepatic glycogen content (P<0.05) in triterpene groups was observed compared with HCD‐fed groups. Furthermore, ACAT gene expression was suppressed compared with hyperlipidemia‐induced groups without triterpenes. It can be concluded that triterpene treatment possesses therapeutic effects on diet‐induced hyperlipidemia by inhibiting the intestinal absorption and storage of cholesterol. Drug Dev Res 68:261–266, 2007. © 2007 Wiley‐Liss, Inc.     

Anti-complementary activity of triterpenoides from fruits of Zizyphus jujuba

In order to determine on the anti-complement activity of triterpenes, following eleven triterpenoides were isolated from the fruits of the Zizyphus jujuba MILL: ceanothane-type triterpenes: colubrinic acid (1), zizyberenalic acid (11); lupane-type triterpenes: alphitolic acid (2), 3-O-cis-p-coumaroyl alphitolic acid (3), 3-O-trans-p-coumaroyl alphitolic acid (4), betulinic acid (7), betulonic acid (9); and oleanane-type triterpenes: 3-O-cis-p-coumaroyl maslinic acid (5), 3-O-trans-p-coumaroyl maslinic acid (6), oleanolic acid (8), oleanonic acid (10). These compounds were examined for their anti-complement activity against the classical pathway of the complement system. Among them, compounds 5, 6, and 8 exhibited significant anti-complement activity with IC(50) values of 101.4, 143.9, and 163.4 microM, respectively, whereas the ceanothane-type and the lupane-type triterpenes were inactive. This suggests that the oleanane-structure plays an important role in inhibiting the hemolytic activity of human serum against erythrocytes.     

Antihistaminic and antieicosanoid effects of oleanolic and ursolic acid fraction from Helichrysum picardii

Helichrysum picardii Boiss. & Reuter is a Mediterranean vegetal species from the Asteraceae family. From the methanolic extract of the aerial flowering parts of this plant, a fraction of two pentacyclic triterpenes has been isolated. Gas chromatography revealed that the triterpene isomers ursolic and oleanolic acids comprised 69% and 29% respectively of the composition of this fraction. The triterpene isomeric fraction was tested in two phagocyte cell systems. It inhibited compound 48/80-induced histamine release from rat peritoneal mast cells in an approximately percentage of 45% at 100 microM and myeloperoxidase secretion from A23187-ionophore-stimulated rat peritoneal leukocytes in a significant manner at doses of 50 and 100 miroM. Furthermore, the triterpene isomers very significantly and dose-dependently inhibited generation of the cyclo-oxygenase metabolite prostaglandin E2 (41% inhibition at 50 miroM) and the 5-lipoxygenase metabolite leukotriene B4 (79% inhibition at 50 microM) from activated rat leukocytes. This anti-eicosanoid activity of the triterpene fraction was more potent than that produced by the pure triterpene oleanolic acid used for comparision, indicating a stronger action of the ursolic acid, the major compound of the isolated triterpene fraction. From these data, it can be suggested that the triterpene isomers oleanolic and ursolic acids present in the medicinal plant Helichrysum picardii contribute to the anti-inflammatory profile of this vegetal species.     

远志木部及皮部总皂苷含量比较分析

目的：对远志原药材木部及皮部中总皂苷含量进行比较分析。用以指导临床用药。方法：以齐墩果酸为对照品,香草醛-高氟酸为显色剂,采用紫外－可见分光光度法在575nm处测定远志总皂苷含量。结果：以此方法对远志木部及皮部中总皂苷含量进行比较、分析,为远志皮部总皂苷含量为3．33％、木部总皂苷含量为0．69％。结论：远志中皮部总皂苷含量远远大于木部,远志总皂苷主要存在于根皮中。因此．在提取远志中的总皂苷时,应选用根皮为原料药材。     

Oleanonic acid, a 3-oxotriterpene from Pistacia, inhibits leukotriene synthesis and has anti-inflammatory activity.

One of the best known bioactive triterpenoids is oleanolic acid, a widespread 3-hydroxy-17-carboxy oleanane-type compound. In order to determine whether further oxidation of carbon 3 affects anti-inflammatory activity in mice, different tests were carried out on oleanolic acid and its 3-oxo-analogue oleanonic acid, which was obtained from Pistacia terebinthus galls. The last one showed activity on the ear oedema induced by 12-deoxyphorbol-13-phenylacetate (DPP), the dermatitis induced by multiple applications of 12-O-tetradecanoyl-13-acetate (TPA) and the paw oedemas induced by bradykinin and phospholipase A2. The production of leukotriene B4 from rat peritoneal leukocytes was reduced by oleanonic acid with an IC50 of 17 microM. Negligible differences were observed in the response of both triterpenes to DPP, bradykinin, and phospholipase A2, while oleanonic acid was more active on the dermatitis by TPA and on the in vitro leukotriene formation. In conclusion, the presence of a ketone at C-3 implies an increase in the inhibitory effects on models related to 5-lipoxygenase activity and on associated in vivo inflammatory processes.     

Further triterpenes from Melissa officinalis L (author's transl)

Further Triterpenes from Melissa officinalis L. Melissa off. contains as further triterpenes pomolic acid, three 2-hydroxy- and one 29-hydroxytriterpenic acid. The root contains betulinic acid, betulin and three 2-hydroxytriterpenic acids. The configuration of the 2,3-dihydroxytriterpenic acids was proved by synthesis of the four isomeric 2-hydroxy-derivatives of oleanolic acid, ursolic acid and α-amyrin.