Microwave-assisted Extraction of Rutin and Quercetin from Flos Sophorae

Three microwave-assisted extraction(MAE) procedures were studied. The first procedure was household microwave oven dynamic extraction(HMODE). The second procedure was special microwave oven bath extraction(SMOBE). The third procedure was microwave resonant cavity dynamic extraction(MRCDE). The results obtained by the three microwave-assisted extraction procedures were compared with those obtained by using traditional Soxhlet extraction. The results indicate that the MAE not only took a shorter time, but also simplified the procedure, and made the extraction a higher yield. At the same time the results obtained by the three MAE procedures were also compared with each other.

Relaxant Effects of Quercetin and Rutin on Human Isolated Bronchus

Increasing epidemiological evidence supports the view, that quercetin has protective roles in a multitude of disease states in human who have a high intake of polyphenols. To investigate the ability of quercetin and its rutinoside, rutin, to modulate the relaxation of human airways smooth muscle and to determine the mechanism (s) of such relaxation, isolated human bronchus rings were suspended in individual organ baths, precontracted with acetylcholine or with histamine and the relaxing effects of quercetin and rutin were determined by measurement of isometric tension. Quercetin induced concentration-dependent relaxant responses on acetylcholine or histamine precontracted human bronchial rings and with almost equal effectiveness. In terms of potency (pD2) and efficacy (Emax), quercetin is more potent than rutin on relaxant responses of human bronchus. K+ and Ca2+ concentration-dependent contraction curves were inhibited after incubation with increasing concentrations of quercetin. Quercetin potentiated in a concentration-dependent manner the relaxant effects of isoprenaline or sodium nitroprusside. Rutin had no effect on K+-induced contraction and on relaxant activity of isoprenaline or sodium nitroprusside. Our results suggest that the bronchodilator effects of quercetin are modulated by an increase in cyclic nucleotide levels as well as an alteration in availability of Ca2+ to the contractile machinery.

Study of inducibility and mechanism of phase 2 enzymes by quercetin and rutin

Objective The increasing recognition of the role for oxidative stress in hepatic disorders has led to extensive investigation on the protection by exogenous antioxidants against hepatic injury.In this study,we choose two typical polyphenol,quercetin and rutin,to investigate the mechanism of induction of cellular antioxidants and phase 2 enzymes in human HepG2 cells.Methods The HepG2 cells were treated with various concentrations of quercetin and rutin for 6 h and 24 h.The activities of NAD(P)H:quinone oxidoreductase(NQO1)in HepG2 cells were measured by 2,6-dichloroindophenol reduction method.The content of superoxide dismutase(SOD)was determined with the method of chemical colorimetry.The protein expressions of NQO1 and NF-E2-related factor 2(Nrf2)in HepG2 cells were detected by Western blotting.Results Incubation of HepG2 cells with quercetin and rutin resulted in a marked concentration-and time-dependent induction of a number of cellular antioxidants and phase 2 enzymes,including NQO1,SOD.Quercetin and rutin treatment of HepG2 cells also caused increase in protein expressions of NQO1 and Nrf2.Conclusions This study demonstrates that a series of phase 2 enzymes in HepG2 cells can be induced by quercetin and rutin in a concentration-and time-dependent fashion by upregulation the protein expression of nrf2.

Cytoprotective effects of quercetin and its sugar-containing natural congeners in cultured HEK293 cells injured by anoxia/hypoglycemia and the structure-effect relationship thereto

Objective To comparatively study anti-free radical and cytoprotective effects of quercetin(Q)and its monoglucoside isoquercetin(I),diglucoside rutin(R),which differs only in glycosyl-substitution at C-3 position of the molecules,using anoxia/hypoglycemia-induced cell injury model and thereby to explore the structure-effect relationship thereto.Methods The cell injury model was established by HEK293 cells cultured in vitro with Na2S2O3 plus sugar-free Earle's fluid as incubation medium;Cell survival rate(CSR),total antioxidant capacity(TAC),SOD and LDH levels were determined;The effect intensity of the 3 flavonoids were compared by means of IC50,the concentration required to achieve 50% inhibition of the changes in the above indices in injured cells.Results Q,I and R all concentration-dependently elevated CSR,TAC and SOD,and reduced LDH level;the all of IC50s for the above indices were ranked in order of IC50,Q<IC50,I<IC50,R,namely,the effect intensity should be Q>I>R.Conclusions The 3 structurally similar flavoloids all have significant and concentration-dependent anti-free radical and cyto-protective effects with the intensity being in order of aglycone>monoglucoside>diglucoside;the substitution of-OH by sugar group at C-3 position of flavoloids and increase in the sugar-substituent number are associated with the effect intensity reduced;namely,the intensity of these effects of flavonoids is negatively related the substutution by sugar group at C-3 position.

Anti-free radical and cytoprotective effects of quercetin and its sugar-containing natural congeners in cultured HEK293 cells injured by anoxia/hypoglycemia and the structure-effect relationship thereto

Objective To comparatively study anti-free radical and cytoprotective effects of quercetin(Q)and its monoglucoside isoquercetin(I),diglucoside rutin(R),which differs only in glycosyl-substitution at C-3 position of the molecules,using anoxia/hypoglycemia-induced cell injury model and thereby to explore the structure-effect relationship thereto.Methods The cell injury model was established by HEK293 cells cultured in vitro with Na2S2O3 plus sugar-free Earle's fluid as incubation medium.Cell survival rate(CSR),total antioxidant capacity(TAC),SOD and LDH levels were determined.The effect intensity of the 3 flavonoids was compared by means of IC50,the concentration required to achieve 50% inhibition of the changes in the above indices in injured cells.Results Q,I and R all concentration-dependently elevated CSR,TAC and SOD and reduced LDH level.The all of IC50s for the above indices were ranked in order of IC50,Q<IC50,I<IC50,R,namely,the effect intensity should be Q>I >R.Conclusions The 3 structurally similar flavoloids all have significant and concentration-dependent anti-free radical and cyto-protective effects with the intensity being in order of aglycone>monoglucoside>diglucoside;the substitution of-OH by sugar group at C-3 position of flavoloids and increase in the sugar-substituent number are associated with the effect intensity reduced;namely,the intensity of these effects of flavonoids is negatively related the substutution by sugar group at C-3 position.

Evaluation of the Mutagenicity Potential of Trace-Rutinosidase Variety of Tartary Buckwheat (Fagopyrum tataricum Gaertn.) Using the Ames Test

To ensure the safety of “Manten-Kirari”, a non-bitter and trace-rutinosidase variety of Tartary buckwheat, we evaluated its mutagenic activity in a bacterial reverse mutagenicity assay, the Ames test. Salmonella typhimurium TA100, TA1535, TA98, TA153, and Escherichia coli WP2 uvrA were employed as test bacteria. The flour of “Manten-Kirari” was dissolved at 12 - 50,000 μg/mL in DMSO and investigated. The number of revertants did not differ compared to the negative control for all concentrations tested, whereas that in the positive control, the number of revertants was increased with or without metabolic activation for each bacterial strain tested. These results suggested that the flour of the Tartary buckwheat “Manten-Kirari” was not genotoxic.

HPLC法同时测定杠板归不同部位中5种成分的含量

目的建立高效液相色谱法同时测定杠板归不同部位(茎、叶、根)中5种成分的含量。方法采用Luna?-C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸水为流动相梯度洗脱,流速为1.0 mL/min,检测波长为320 nm,柱温为30℃,测定杠板归不同部位中绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素的含量。结果绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素分别在0.046~4.6μg/mL,0.212~21.2μg/mL,0.055~5.5μg/mL,0.084~8.4μg/mL,0.114~11.4μg/mL范围内质量浓度与峰面积呈线性关系良好(r>0.9990),平均加样回收率为97.65%~102.09%,RSD为1.76%~2.44%。不同部位中5种成分含量差异较大,绿原酸和咖啡酸2个成分在叶中含量高,且远远高于根和茎中的含量;芦丁和阿魏酸主要存在于茎和根中,叶中未检测到;槲皮素在茎中含量最高,叶中次之,根中最低。结论建立的HPLC方法准确可靠、重复性好、专属性强,可用于杠板归不同部位中5个成分含量的同时测定。

不同发酵工艺对浙产六神曲中5种成分含量变化的影响

目的建立同时测定浙产六神曲中苦杏仁苷、香草酸、芦丁、阿魏酸、槲皮素的高效液相色谱法,并考察不同发酵条件对浙产六神曲中5个成分含量变化的影响。方法对同一批次六神曲样品,通过设置不同的发酵条件(温度为25℃、30℃、35℃,相对湿度为60%、70%、80%、90%)进行发酵并分别取样。色谱柱为Chrom Core AR C18柱(4.6 mm×250 mm,5μm);以乙腈为流动相A,0.08%磷酸水溶液为流动相B,进行梯度洗脱;流速为1.0 ml/min,柱温为30℃,检测波长220 nm。分析苦杏仁苷、香草酸、芦丁、阿魏酸、槲皮素5种成分的变化。结果苦杏仁苷、香草酸、芦丁、阿魏酸、槲皮素在选定质量浓度范围内线性关系、重复性、稳定性良好,平均加样回收率均达到要求。浙产六神曲最佳发酵工艺参数为温度30℃,相对湿度80%,发酵时间为3 d。结论本研究建立的方法操作简便、结果准确、重复性及稳定性均较好,可用于浙产六神曲的质量控制。浙产六神曲最佳发酵工艺参数为温度30℃,相对湿度80%,发酵时间为3 d。

3种中药成分对玉米赤霉烯酮致小鼠肝脏损伤的缓解作用

【目的】探究中药对玉米赤霉烯酮(zearalenone, ZEA)致动物肝损伤的缓解作用。为临床研发新中兽药和饲料添加剂提供理论依据。【方法】将63只8周龄昆明雌鼠随机分为7组,每组9只。对照组,上、下午灌胃生理盐水各1次;ZEA组,上午灌胃20 mg/kg ZEA,下午灌胃生理盐水;槲皮素、山奈酚、芦丁和成分复方组为中药成分治疗组,上午灌胃20 mg/kg ZEA,下午分别灌胃50 mg/kg槲皮素、50 mg/kg山奈酚、100 mg/kg芦丁、100 mg/kg成分复方(V_((槲皮素))∶V_((山奈酚))∶V_((芦丁))=1∶2∶2);阳性对照组,上午灌胃20 mg/kg ZEA,下午灌胃100 mg/kg维生素E;每只每次均灌胃0.2 mL,连续7 d。于第8天称量各组小鼠体重,眼球摘除采血,分离血清,脱颈处死小鼠,剥离肝脏并称重,计算肝脏指数,检测血清肝脏功能、抗氧化酶活性和肝脏炎症指标。【结果】生长发育检测结果显示,与对照组相比,ZEA组小鼠体重降低、肝脏重量升高(P<0.05);与ZEA组相比,芦丁和成分复方组体重升高(P<0.05)。血清肝脏功能检测结果,与对照组相比,ZEA组血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、乳酸脱氢酶(LDH)活性均升高(P<0.05);与ZEA组相比,中药成分和维生素E治疗组各个酶活性均降低(P<0.05)。肝脏氧化应激检测结果显示,与对照组相比,ZEA组肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性、谷胱甘肽(GSH)含量和总抗氧化能力(T-AOC)均降低(P<0.05),丙二醛(MDA)和LDH含量升高(P<0.05);与ZEA组相比,中药成分和维生素E治疗组SOD、CAT活性、GSH含量和T-AOC均升高(P<0.05),MDA和LDH含量降低(P<0.05)。炎症因子检测结果,与对照组相比,ZEA组机体肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)和IL-6含量升高、IL-10含量降低(P<0.05);与ZEA组相比,中药成分和维生素E治疗组机体TNF-α、IL-1β和IL-6含量降低(P<0.05),槲皮素和维生素E治疗组肝脏IL-10含量升高(P<0.05)。【结论】3种中药成分槲皮素、山奈酚、芦丁和维生素E均可缓解ZEA致小鼠肝脏的损伤,山奈酚效果最佳。

黄连汤化学成分研究

黄连汤出自张仲景的《伤寒论》,由黄连、炙甘草、干姜、桂枝、人参、半夏、大枣组成。在中医临床中广泛用于胸中有热,胃中有寒引起的胸中烦热,欲呕吐,腹中疼痛。综述了黄连汤组成中药的化学成分,并采用硅胶、聚酰胺柱层析及重结晶方法对黄连汤的化学成分进行分离纯化,根据理化性质和光谱数据分析鉴定得到9个化合物,分别是:槲皮素(quercetin,1)、芦丁(rutin,2)、胡萝卜甙(daucosterol,3)、齐墩果酸(oleanolic acid,4)、甘草次酸(glycyrrhetinic acid,5)、小檗碱(berberine,6)、大黄酚(chrysophanol,7)、氧化小檗碱(oxyberberine,8)、5-羟甲基糠醛(5-hydroxymethylfurfural,9)。以上化合物均为首次从该复方中分离得到。

女贞子中多种有效化学成分的同步测定方法研究

建立一种高效液相色谱法同步分析女贞子中咖啡酸、芦丁、槲皮素和熊果酸四种有效化学成分的方法.使用Waters SymmetryshieldTMRP18(4.6×250 mm,5μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相梯度洗脱,体积流量1.0 mL/min,检测波长范围190~400 nm,柱温30℃,进样量10μL.在此色谱条件下,芦丁、槲皮素、咖啡酸和熊果酸四种成分分离效果良好,在40~250μg/mL、40~400μg/mL、80~700μg/mL和40~400μg/mL范围内各相关系数均大于0.999,不同浓度条件下芦丁、槲皮素、咖啡酸和熊果酸日内精密度试验的平均RSD值分别为0.41%、0.61%、0.61%、0.86%,日间精密度试验的平均RSD值分别为0.43%、0.52%、0.68%、0.83%,稳定性试验的RSD值分别在0.17%~0.67%、0.54%~0.80%、0.32%~0.87%、0.36%~1.29%,平均回收率分别为100.81%、97.68%、102.83%、100.01%.该方法精密度高、稳定性良好,可以对女贞子中芦丁、槲皮素、咖啡酸和熊果酸四种有效成分进行同步准确测定,研究结果可为四种有效化学成分的深入研究以及女贞子的开发应用提供研究基础.

槲皮素-3-O-乙酸-(4-(3H-1,2-二硫杂环戊烯-3-硫酮))-苯酯的合成

以芦丁为起始原料,以5-(4-羟基苯基)-3H-1,2-二硫杂环戊烯-3-硫酮作为H_(2)S供体,合成一种具有治疗糖尿病且促进伤口愈合功能的药物槲皮素-3-O-乙酸-(4-(3H-1,2-二硫杂环戊烯-3-硫酮))-苯酯。整个工艺路线包括取代、水解、williamson醚化、水解和酯化等5步反应,总产率为6.4%。各步产物经MS、^(1)H NMR分析表征。该合成路线具有原料廉价易得、反应条件温和、操作简便的优点。

高效液相色谱法同时测定固体饮料中没食子酸、芦丁、鞣花酸和槲皮素的含量

目的建立高效液相色谱法(high performance liquid chromatography,HPLC)同时测定固体饮料中没食子酸、芦丁、鞣花酸和槲皮素含量的方法。方法样品经15%二甲亚砜(dimethyl sulfoxide,DMSO)甲醇溶液萃取,离心后取上清液,经0.45μm微孔滤膜过滤后用高效液相色谱仪分析。选用色谱柱Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm);流动相A为乙腈,流动相B为0.2%磷酸水溶液,梯度洗脱;流速为1.0 mL/min;柱温为25℃;进样体积5μL;检测波长没食子酸为273 nm,芦丁、鞣花酸和槲皮素为254 nm,运行时间为55 min。结果没食子酸、芦丁、鞣花酸和槲皮素分别在0.00072~0.02150 mg/mL(r^(2)=0.9953)、0.00380~0.15000 mg/mL(r^(2)=0.9998)、0.00033~0.01300 mg/mL(r^(2)=0.9995)、0.00062~0.02500 mg/mL(r^(2)=0.9999)范围内线性关系良好;没食子酸、芦丁、鞣花酸、槲皮素的加标回收率在95.7%~109.7%之间,相对标准偏差在0.7%~2.2%之间。本方法中检出限和定量限:没食子酸分别为11.0μg/g和33.0μg/g;芦丁分别为6.6μg/g和20.0μg/g;鞣花酸分别为1.5μg/g和5.2μg/g;槲皮素分别为3.5μg/g和12.0μg/g。结论本方法前处理方法简单,检测灵敏度高,稳定性好,并且可以有效地把食品样品中没食子酸、芦丁、鞣花酸和槲皮素几种植物化学物质同时准确检测,可以作为成分复杂的固体饮料中多个植物化学物的质量控制的参考。

HPLC-MS/MS同时测定感冒安颗粒中5种黄酮成分的含量

目的建立HPLC-MS/MS方法测定感冒安颗粒中5种黄酮成分含量。方法色谱条件:色谱柱为Kromasil C_(18)柱(4.6 mm×150 mm×4.6 mm,5μm,100Å),流动相为甲醇(A)-0.1%甲酸(B),梯度洗脱(0~20 min,35%A;20~40 min,45%A),流速为1.0 ml/min,柱温为25℃;质谱条件:电喷雾电离源(ESI源)负离子扫描,多反应监测(MRM)模式检测。5种黄酮检测的离子对:芦丁m/z 609.1→300.1、金丝桃苷和异槲皮苷m/z 463.0→300.1、槲皮素m/z 301.0→151.0、木犀草素m/z 285.0→132.9。结果5个成分在一定浓度范围内与峰面积具有良好的线性关系,相关系数r均>0.9991;平均加样回收率在100.63%~102.81%之间,RSD范围在0.67%~2.07%。10批感冒安颗粒中芦丁、金丝桃苷、异槲皮苷、槲皮素和木犀草素的含量测定结果分别为32.23~479.83、0.291~1.825、11.44~20.54、6.32~18.41、3.46~6.51μg/g。结论建立的HPLCMS/MS法灵敏度高、专属性强,适合同时测定感冒安颗粒中5种黄酮类成分的含量。

槲皮素体内外抗氧化作用的比较研究

目的 :测定槲皮素的体外总抗氧化力 ,进一步观察槲皮素灌胃后大鼠外周血总抗氧化力的变化 ,并与芦丁、维生素C、维生素E相比较。方法 :总抗氧化力采用Fe3 + 还原法 ,槲皮素、芦丁分析采用紫外分光光度法及高效液相色谱法。结果 :相同浓度条件下槲皮素的体外总抗氧化力显著强于芦丁 ,与传统的抗氧化剂维生素C、维生素E相当。槲皮素 4 0mg/kg灌胃 1h后大鼠外周血总抗氧化力及槲皮素含量 (紫外分光光度法 )较灌胃前升高最为明显。维生素C也有显著提高外周血总抗氧化力的作用 ,芦丁与维生素E未表现出显著作用。血浆高效液相分析表明槲皮素灌胃后未出现明显的槲皮素吸收峰 ,而与其峰相邻的两个未知峰的面积增大。结论 :槲皮素的体外抗氧化作用强于芦丁 ,与传统的抗氧化剂维生素C、维生素E相当 ;槲皮素吸收后经代谢形成衍生物 。

槲皮素及异槲皮素、芦丁抗自由基活性的比较研究

目的评价3种黄酮类同系化合物槲皮素及其单糖苷异槲皮素和双糖苷芦丁的体外抗自由基作用,进一步研究其构效关系。方法采用H2O2/Fe2+体系,通过Fenton反应生成羟自由基(·OH);采用邻苯三酚自氧化法产生超氧阴离子(O2-)。利用·OH造成肝细胞脂质过氧化以及红细胞膜破裂,观察受试化合物的保护作用,并计算IC50。结果在以上4种体外模型中,3种药物均表现出极强的清除自由基能力。在·OH以及O2-清除实验中,抗氧化活性顺序为芦丁>异槲皮素>槲皮素;在抑制肝匀浆脂质过氧化及红细胞膜破裂实验中,抗氧化活性顺序为槲皮素>异槲皮素>芦丁。结论3种受试黄酮类化合物均具有十分强大的自由基清除作用,且呈现明显的剂量依赖性和糖基结构依赖性,在不同的实验体系中其抗自由基活性强弱顺序不同。

槲皮素、芦丁及葛根素抑制LDL氧化修饰作用的研究

目的:研究黄酮类中药槲皮素、芦丁及葛根素抑制低密度脂蛋白(LDL)氧化修饰的作用。方法:按一次性密度梯度超速离心法分离人血清LDL,分别加入5.0μmol.L-1槲皮素、芦丁及葛根素与LDL温育3 h,然后利用Cu2+介导体外氧化修饰脂蛋白的方法,观察LDL氧化修饰不同时间,其A234,相对电泳迁移率(REM),硫代巴比妥酸反应物质(TBARS)及载脂蛋白羰基含量的动力学改变,并与对照组比较。结果:Cu2+介导LDL氧化修饰时,在2,4,6,8,12,24 h后,其A234,REM,TBARS及载脂蛋白羰基含量均逐步增加;加入5.0μmol.L-1槲皮素、芦丁及葛根素的实验组,Cu2+介导LDL的氧化修饰分别延迟2~6 h,2 h和2 h,其A234,REM,TBARS及载脂蛋白羰基含量较对照组降低;槲皮素实验组分别较对照组降低27.7%~49.6%,24.1%~38.6%,19.8%~34.3%及36.4%~56.8%;芦丁实验组分别降低12.8%~39.3%,15.7%~32.0%,19.0%~28.1%及12.8%~50.3%;葛根素实验组分别降低3.3%~19.2%,7.0%~22.5%,19.5%~22.8%及8.6%~47.0%。结论:槲皮素、芦丁及葛根素均可抑制Cu2+诱导的LDL的氧化修饰,且槲皮素的抗氧化作用较芦丁及葛根素强。

云南普洱茶中黄酮含量的测定及槲皮素、芦丁的提取分离

研究了部分云南普洱茶中黄酮类化合物的含量及槲皮素、芦丁的提取分离方法。实验结果表明,新工艺原料黄酮含量最高,达24.576mg/g,新工艺特级最低,仅有3.660mg/g。从中选取黄酮含量较高的三个样品进一步提取分离槲皮素、芦丁,得到了纯度在30%以上的粗制品。

不同肠段对槲皮素、芦丁吸收的比较研究

为了比较不同肠段对槲皮素及芦丁的吸收情况 ,选用Wistar大鼠 72只 ,随机分为对照组、槲皮素组及芦丁组 ,每组分别进行十二指肠、空肠、回肠及结肠灌流。灌流液分别为 1%的二甲基亚砜溶液 (溶剂 )、2 0 μmol L槲皮素、芦丁溶液。灌流结束后 ,采用高效液相色谱仪分析灌流液中槲皮素或芦丁的含量。结果发现 :十二指肠、空肠、回肠、结肠对槲皮素及芦丁均有吸收 ,芦丁的吸收率显著低于槲皮素。槲皮素单位长度吸收率 :结肠 >回肠 >十二指肠 >空肠 ,单位面积吸收率 :回肠 >空肠 >十二指肠 >结肠。槲皮素肠道灌流后 ,高效液相色谱图中出现新的未知峰 ,推测其为肠粘膜代谢后形成的衍生物重新分泌进入肠腔所致。

山豆根化学成分研究

利用多种柱层析方法,从山豆根中分得5个化合物。经过理化及波谱分析,分别鉴定为L 高丽槐素( 1)、红车轴草苷( 2 )、槲皮素( 3)、芦丁( 4 )和异鼠李素 3 芸香糖甙( 5 )。其中化合物3。