基于UPLC-Q/TOF MS及网络药理学的丹参川芎嗪注射液抗血瘀活性成分和机制研究

运用网络药理学方法研究丹参川芎嗪注射液抗血瘀的活性成分及机制,采用UPLC-Q/TOF MS技术测定其化学成分,通过UNIFI天然产物分析平台,对比各成分的精确分子质量、保留时间及质谱碎片离子信息,并分析鉴定其结构.利用里宾斯基五规则筛选出全成分中的活性成分,运用TCMSP、DisGeNET、DAVID等数据库预测活性...     

高效液相色谱法测定凉茶中绿原酸、新橙皮苷、甘草酸的含量

目的：建立凉茶中绿原酸、新橙皮苷、甘草酸的高效液相色谱测定方法。方法：采用ZORBAX SB-C18柱（4.6×250 mm,5μm）,以乙腈-0.1%磷酸溶液为流动相,梯度洗脱,流速1.0mL/min;进样量10μL。结果：绿原酸、新橙皮苷、甘草酸的线性范围均为0.10～50mg/L,样品的平均加标回收率范围分别为90.1%-94.0%、97.8%-115%、78.7%-91.9%,相对标准偏差分别为0.60%、1.16%、0.67%。该方法操作简便、快速、准确,应用于凉茶中绿原酸、新橙皮苷、甘草酸的分析,取得了较好的结果。     

亚麻子发芽前后化学成分及抗氧化活性的研究

采用高效液相色谱-二级串联质谱(HPLC/MS²)技术对不同发芽时间(0~8天)的亚麻子芽的化学成分进行鉴定。利用紫外分光光度法对亚麻子发芽前后的总黄酮和总多酚类成分进行定量分析,并采用铁离子还原能力法(FRAP)、自由基清除能力法(DPPH)和氧自由基吸收能力法(ORAC)对亚麻子芽的抗氧化活性进行评价。采用Phenomenex ODS C18色谱柱(4.6 mm×250 mm×5 μm),以甲醇-0.2%乙酸水溶液为流动相进行线性梯度洗脱,流速0.75 mL/min,柱温25 ℃,全波长扫描紫外检测器(DAD),检测波长为280 nm;采用电喷雾离子源(ESI)在正、负离子模式下进行检测。相比于亚麻子,在亚麻子芽样品中鉴定出12种新化合物,包括糖类、嘌呤类、核苷类和黄酮类化合物。对这12种化学成分进行定量和抗氧化活性分析,结果表明,随着发芽时间的增加,亚麻子芽中的总黄酮、总多酚类成分的量以及抗氧化活性逐渐增加。该方法可为亚麻子作为保健品以及功能性食品的原料提供理论依据。     

大青叶化学成分及抗氧化活性研究

采用高效液相色谱-大气压化学电离质谱法（HPLC-APCI/MS）和直接进样的电喷雾串联质谱法（ESI-MS²）分析大青叶乙酸乙酯、正丁醇和95%乙醇提取物的化学成分。采用Kromasil C18分析柱（4.6 mm×250 mm×5 μm），以0.15%乙酸/水-甲醇为流动相进行线性梯度洗脱，流速0.8 mL/min，柱温25 ℃；采用全波长扫描紫外检测器,检测波长分别为254 nm和265 nm；采用大气压化学电离源和电喷雾电离源，在正、负两种离子模式下进行检测。结果表明，在大青叶乙酸乙酯、正丁醇和95%乙醇提取物中共鉴别出39种化合物，包括生物碱类、有机酸类、核苷嘌呤类、氨基酸类、黄酮类、糖类、卟啉类以及含硫类成分化合物。采用铁离子还原能力法（FRAP）对大青叶的抗氧化活性进行评价，并用自由基清除能力法验证FRAP法的测试结果。抗氧化活性评价结果表明，大青叶抗氧化活性部位主要集中在极性较低的化学组分中，吲哚类、奎唑酮类和有机酸类化合物是其主要有效成分。该方法快捷、灵敏，适用于中药化学成分分析和抗氧化活性评价。     

Hela细胞增殖抑制模型结合离心超滤质谱筛选中药抗癌药物

采用基于MTT比色法的Hela细胞增殖抑制模型评价中药提取物的抗癌活性,并利用离心超滤质谱方法对活性最强的中药提取物中抗癌活性成分进行筛选和鉴定。结果表明,长春花提取物对Hela细胞增殖的抑制活性最强,与阳性对照药阿霉素相当,且抑制活性与给药浓度呈正相关。延胡索提取物在3个不同给药浓度下也可以显著抑制Hela细胞的增殖。三尖杉、黄连和喜树果提取物在高浓度时能够显著抑制Hela细胞的增殖,而刺五加提取物则没有明显抑制作用。利用离心超滤质谱从长春花提取物中筛选得到2种存活素基因启动子片段结合剂和6种钙调蛋白结合剂,经多级串联质谱分析鉴定,分别为蛇根碱、鸡骨常山碱、它波宁、环氧长春碱、长春新碱和21’-氧代环氧长春碱。本研究可为中药抗癌药物的筛选提供一种简便、快速的方法。     

复合营养素制剂人体抗氧化功能试验研究

目的探讨以番茄红素与葡萄籽提取物为主要原料制成的复合营养素制剂对人体抗氧化方面的作用。方法将120例符合要求的健康志愿者随机分为试食组和对照组,试食组连续服用复合营养素制剂样品120 d,对照组服用安慰剂。试食前后各测定一次血清中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-PX)活性和安全性指标。结果试食后,试食组的MDA含量平均下降0.33±0.14 nmol/mL,下降率为8.07%,自身及组间比较的差异具有统计学意义(p0.01);SOD活性平均升高7.98±2.49 U/mL,上升率为8.71%,自身及组间比较的差异具有统计学意义(p0.01);GSH-PX活性平均升高9.42±6.07 U/mL,上升率为7.71%,自身及组间比较的差异具有统计学意义(p0.01),各项安全性指标试验前后均无明显变化。结论以番茄红素与葡萄籽提取物为主要原料制成的复合营养素制剂对人体具有抗氧化功能的作用。     

现代色谱联用技术在中药质量控制中的应用

以作者近几年研究进展为例探讨现代色谱联用技术特别是液相色谱-质谱联用技术在中药质量控制中的应用。重点讨论中药全成分快速表征创新策略和方法，以及在中药同属药材的鉴别、药材初加工评价、炮制品区分、配方颗粒质量评价、活性成分资源寻找和毒性成分筛选等方面的应用。     

亲和超滤质谱技术在中药活性成分筛选中的研究进展

亲和超滤质谱技术是20世纪90年代中期发展起来的一种快速、简单、有效的药物小分子发现模式。该技术利用配体与受体之间特异性结合,通过超滤装置快速筛选活性小分子化合物,再结合液相色谱 质谱联用技术（LC-MS）,鉴定活性成分结构。亲和超滤质谱技术集药物活性成分筛选、结构鉴定于一体,尤其适用于从复杂体系中筛选潜在的活性小分子化合物。近年来,针对中药发挥药理作用具有多组分、多靶点的重要特点,亲和超滤质谱技术已被广泛用于从中药提取物中筛选与特定蛋白靶点相结合的小分子活性物质,对阐明中药药效的物质基础和以活性成分作为先导化合物的新药开发具有重要意义,是对传统药物发现方法的有利补充。本工作综述了该技术在中药活性成分筛选中的原理、特点、应用进展,以及对未来的展望。     

UPLC-MS-MS测定红茶中的茶黄素含量

建立了一种快速测定红茶中4种茶黄素(茶黄素、茶黄素-3-没食子酸酯、茶黄素-3'-没食了酸酯和茶黄素-3,3'-双没食了酸酯)的超高效液相色谱串联质谱的分析方法。样品用70%的甲醇水浴提取,采用Waters ACQUITY BEH C18色谱柱分离,串联四级杆质谱多反应离子监测,外标法定量;色谱系统流速为250μL/min,柱温为30℃,流动相为0.1%甲酸和乙腈,梯度洗脱。结果表明,4种茶黄素在检测范围内线性关系良好,相关系数大于0.995,平均回收率在91.95%~105.37%之间,RSD(n=5)在2.37%~4.03%;方法的检出限在13.2~19.1 ng/m L,方法的定量限在39.7~52.6 ng/m L。所测红茶茶黄素含量为3.32 mg/g、茶黄素-3-没食子酸酯含量为1.96 mg/g、茶黄素-3'-没食了酸酯含量为1.48 mg/g、茶黄素-3,3'-双没食了酸酯0.70 mg/g,茶黄素总量为0.75%。该检测方法能快速检测红茶中的茶黄素含量,能有效分析评价红茶的品质,合理控制红茶生产质量。     

高效液相色谱法测定催泪喷射器的成分含量

利用简单流动相组成和商品柱,建立了测定催泪喷射器中有效成分的反相高效液相色谱法.色谱柱为Kromasil KR-1005C18(250 mm×4.6 mm,5μm),流动相为甲醇:水(80:20,V/V),流速0.8 mL/min,柱温30℃,紫外检测波长为205 nm.能够明确地分辨催泪喷射器内容物中主要成分:邻氯苯亚甲基丙二腈.该方法简便快捷,线性关系良好,结果准确可靠,对修改催泪喷射器的成分含量检测的标准具有重要作用.     

Influence of aldehydes in make-up oils on antioxidation properties

The addition of more antioxidant to aged oils is not always effective in preventing autoxidation. To investigate this phenomenon, a solution of antioxidant and model oxide in hexadecane was examined by means of the induction period method. Aldehydes were found to be the most active species for reducing the antioxidation properties of zinc dialkyldithiophosphate (ZDTP), 2,6-di(tert. butyl)-4-methylphenol (DBPC), and N-phenyl-1-naphtylamine (PNA). Carboxylic acids, ethers, alcohols, and ketones to some extent reduced the induction period with these antioxidants. Esters showed almost no influence on reducing their antioxidant properties. Reaction of ZDTP with aldehyde was studied by means of ³¹P-NMR analysis. It was found that (a) aldehydes accelerate conversion of ZDTP, and thereby reduce its antioxidant performance. (b) Conversion of ZDTP in the presence of aldehydes under oxygen at 130° in hexadecane and no reaction was observed under nitrogen at the same temperature. Active intermediates derived by the autoxidation of aldehyde were suspected. (c) Reaction of peroxycarboxylic acid with ZDTP took place smoothly at 0°C under nitrogen to give bis(dialkylthiorophosphoro)disulphide and the corresponding carboxylic acid. Reaction of hydroperoxide with ZDTP took place at room temperature to give oxo-hexa(dialkylphosphorothiorothionato)-tetrazinc. Two mechanisms for decomposition of ZDTP promoted by aldehydes, namely hydroperoxide and peroxycarboxylic acid mechanisms, are proposed. Reaction of peroxycarboxylic acid with DBPC or PNA barely took place at room temperature: for these antioxidants, the peroxycarboxylic acid mechanism would be excluded. Radical species derived by the autoxidation of aldehydes were considered as active intermediates. It is considered that classification of the types of oxides that exist in used oils in the key to optimising the effect of re-added antioxidants.     

Antioxidant activity and rosmarinic acid content of ultrasound-assisted ethanolic extracts of medicinal plants

The interest in finding natural antioxidants for use in food, drugs or cosmetic products has increased concerning the replacement of synthetic antioxidants due to their possible undesirable toxic effects on humans long-term exposed.Ethanolic extracts of Hypericum perforatum (Hypericaceae), Lavandula angustifolia (Lamiaceae), Malva sylvestris (Malvaceae), Melissa officinalis (Lamiaceae), Salvia officinalis (Lamiaceae), Rosmarinus officinalis (Lamiaceae) were obtained under sonication with a yield of 5.9%, 3.8%, 5.8%, 3.8%, 4.2% and 2.3% (w/w), respectively and their antioxidant activity assessed by the TLC-DPPH and DPPH assays. With exception of L. angustifolia and M. sylvestris extracts all the others showed antioxidant activity greater than 70% (at 0.1 mg/mL).The HPLC analysis shows that all tested extracts contain rosmarinic acid and suggest that H. perforatum extract have the highest concentration (2.76 mM) and M. sylvestris extract the lowest one (0.47 mM).In this study, we describe a simple laboratory procedure using an ultrasound-assisted ethanolic extraction from six medicinal plants. This potential didactic approach showed to be a rapid and effective process on preserving the antioxidant activity and rosmarinic acid content. The HPLC system was used to evaluate the ability to assess rosmarinic acid content.     

Computer simulation of turbine oil oxidation. 2: Consumption of a hindered phenol antioxidant in laboratory oxidation tests and in service

The computer simulation study of consumption of di-tert-butyl p-cresol (DBPC) was extended to the oxidation of hexadecane/tetralin model oils under conditions of laboratory oxidation tests and in service. The oxidation lives of the model oils in the ALCOA oxidation test and the ASTM D2272 rotary bomb and D943 turbine oil oxidation tests were successfully predicted from the simulation study. The oxidation behaviour of the model oils is compared to that of a turbine oil used in actual service units. The model oil having 6 mass % of tetralin showed almost identical oxidation behaviour to the turbine oil when the same amount of DBPC was added (0.6 mass %). The calculation shows that the direct oxidation of antioxidants is the main process of antioxidant consumption in the laboratory oxidation tests as well as in the model oxidation system without metal catalysts. In most of the service units which operated at moderate conditions the calculation shows that the direct oxidation of antioxidants is also the main, process. As a consequence, the rate of antioxidant consumption does not depend much on the tetralin content in the model oils. The oxidation behaviour of the model oils in the service units which operate under severe conditions, such as in the high temperature regions of turbines, is calculated by setting a flow reactor model. The results of this calculation show in this case that the turbine oil degradation occurs mostly in the high temperature regions and that the rate of antioxidant consumption increases with an increased amount of tetralin in the model oils. This strong dependence on base oil composition can be attributed to the increased importance of antioxidant consumption through the inhibition of oxidation.     

Coke Formation from Aircraft Turbine Engine Oils: Part I — Deposit Analysis and Development of Laboratory Oil Coking Test

The authors report here the results of a study undertaken to determine how aircraft engine oil degrades to form coke in static films on engine components and the impacts of additives and surface materials upon the coke production processes. The research is presented in two parts. This first paper deals with the initial research to develop a simple laboratory oil-coking test capable of monitoring antioxidant depletion, oil degradation, and coke formation of stressed aircraft turbine engine oils. Details of the analytical chemistry experiments performed on the stressed samples to quantitate the percentage of oil, polymer dissolved in the oil, and insoluble coke are also presented. The second paper (Part 2) (I) deals with more quantitative measurements of the degradation of thin layers of oil on heated surfaces. Effects of different oils and surfaces are explored.

The results presented in Part I show that simple open-vial laboratory tests involving thin films of oil produce polymer and coke similar to the deposits seen on failed face seals taken from operating aircraft engines and isolated from used engine oils. Coking reactions require oxygen to deplete the antioxidant package and to polymerize/decompose the ester basestock of the oil. After the antioxidant package is depleted, the ester basestock polymerizes to form small oil-soluble polymers. As the oil spends additional time at elevated temperature, these polymers increase in molecular weight and change in chemical composition becoming insoluble in the oil, producing deposits. Although the antioxidants inhibit the polymer/coke reactions, the antioxidants are incorporated in the formed deposits increasing the deposit amount. Consequently, these results indicate the proper concentration and choice of antioxidants will allow elimination, not just reduction, of deposits on engine components coated with thin layers of static oil films.     

Computer simulation of turbine oil oxidation. 1: Consumption of a hindered phenol antioxidant in model hydrocarbon systems at 115°C

Computer simulation of the inhibited oxidation of model hydrocarbon systems representing turbine oils of different composition, hexadecane/tetralin mixtures containing di-tert-butyl pcresol (DBPC), has been achieved using the kinetic and mechanistic information obtained from fundamental hydrocarbon autoxidation and inhibition studies. The oxidation life, the time to complete consumption of DBPC and its reactive intermediates, was calculated using previously reported and newly measured absolute rate constants for reaction involved. The results of this calculation of oxidation life were in good agreement with values experimentally determined. The oxidation life has been found to decrease with increasing amount of tetralin in the mixtures. This can be explained by the increased rate of consumption of antioxidants, caused by an increase in the rate of free radical formation, via the direct reaction of hydrocarbons with oxygen, and via the homolytic decomposition of hydroperoxides. The observed decrease in oxidation life, however, is not significant, despite the fact that the addition of tetralin considerably increases the rate of free-radical formation. The relatively low sensitivity of oxidation life to the rate of free-radical formation caused by the addition of an easily oxidisable substrate can be attributed to the occurrence of a direct oxidation of antioxidants. That is, kinetic analysis for the consumption of antioxidants disclosed that antioxidant consumption mostly occurred not through the inhibition of oxidation, but through direct oxidation. The direct oxidation of antioxidants obeys the rate equation -d[AH]/dt = k₆[AH]^1.0[O₂]^1.5 where k₆ can be calculated, at various temperatures, from Arrhenius parameters obtained in this study (log (A/M⁻¹s⁻¹) = 9.6 and Ea = 22.6 kcall mol).     

Evaluation of sodium acetylacetonate as a synergist for arylamine antioxidants in synthetic lubricants

Differential scanning calorimetry (DSC) was employed to evaluate the performance of an ester lubricant containing sodium acetylacetonate. In the isothermal DSC oxidation test, sodium acetylacetonate is an effective synergist for arylamine antioxidant p,p′-dioctyldiphenylamine (DODPA). When combining with arylamine antioxidants, sodium acetylacetonate can also effectively reduce the increases in viscosity and acid number of the ester oil in oxidation–corrosion test. The electron paramagnetic resonance (EPR) results indicated that sodium acetylacetonate can reduce the free radical contents in the oxidized oil with arylamine antioxidants present.     

HPLC-ESI-MS/MS分析黄芪蜜炙前后活性成分差异

建立了HPLC-ESI-MS/MS联用技术分析生黄芪与炙黄芪的成分,并比较两者成分差异。采用Agilent ZORBAX SB-C18色谱柱(4.6 mm×250 mm×5μm),以0.1%甲酸乙腈溶液-0.1%甲酸水溶液为流动相进行梯度洗脱,流速1 mL/min,检测波长203 nm,柱温25℃;使用ESI离子源,分别在正、负离子模式下进行全扫描和二级扫描,质量扫描范围m/z 100～2 000。采用该方法鉴定了生黄芪与炙黄芪中的62种成分,并对黄芪蜜炙前后的成分进行了比较,实验结果可为黄芪炮制方法的选择提供参考依据。     

The response to antioxidants in base oils of different degrees of refining

Base oils with different compositions and different degrees of refining have been subjected to three types of oxidation stability tests in the presence of antioxidants. A higher degree of refining of base oils depletes the natural antioxidants present in oils of lower degree of refining. However, the results of the study show that oxidation stability of highly refined oils, after addition of suitable antioxidant packages, by far outperforms that of the oils of lower degree of refining. This is true regardless of the dominating hydrocarbon type in the oil, or its feedstock origin. The results also show that in general, the viscosity index of a base oil, or any other simple physical property, has no direct correlation to oxidation stability or response to antioxidants. To analyse the response to antioxidants, a concept called oxidation stability response is introduced. A novel method for determining the oxidation onset temperature in differential scanning calorimetry measurements by inverse derivatives is introduced. Copyright © 2007 John Wiley & Sons, Ltd.     

中药陆英高效液相色谱指纹图谱的研究

目的：建立中药陆英高效液相色谱指纹图谱,为科学评价及有效控制其质量提供可靠方法。方法：采用高效液相色谱法,色谱柱为Kromasil C18（250mm×4．6mm,5μm）柱,流动相为乙腈-0．2％磷酸水溶液（80：20）,光电二极管阵列检测器,检测波长215nm,流速0．8mL／min,柱温30℃。结果建立了中药陆英的指纹图谱,确定了10个共有峰,各陆英样品指纹图谱与对照指纹图谱相似度均在0．9以上,可用于陆英药材的定性鉴别。结论所建立的HPLC指纹罔谱具有良好的精密度、重现性和稳定性,可以作为陆英质量评价的主要依据之一,同时为深入研究中药陆英的质量标准提供了实验基础。