CZE法鉴别测定苦豆草药材中多种生物碱成分

建立了一种同时在苦参药材中鉴别9种生物碱并测定其中6种生物碱含量的毛细管区带电泳法.即用未涂渍熔融石英毛细管柱50μm×41.5cm(有效长度33cm),以0.3mol/L Tris水溶液一正丙醇(ⅥV=60/40,pH=7.0)为运行缓冲液,氯丙那林为内标,在检测波长210nm、运行电压30kV、50mbar压力下进样10s、运行温度25℃和分析时间40min进行毛细管区带电泳.结果表明,宁夏市售苦豆草中含有野靛碱、莱蔓碱、槐安碱、苦参碱、槐定碱、槐果碱、苦豆碱、氧化槐果碱和氧化苦参碱,其中苦参碱、槐果碱、苦豆碱、氧化槐果碱和氧化苦参碱的含量接近,槐定碱的含量明显高于它们.本法灵敏、准确、专属性好,适用于苦豆草药材中生物碱成分的鉴别和含量测定.     

二-(2-乙基己基)磷酸/正辛醇反胶束萃取L-辛弗林

利用二-(2-乙基己基)磷酸(P204)/正辛醇反胶束从枳实粗提物中萃取L-辛弗林。 研究了萃取机理和水相pH值、含水量W0、阳离子浓度、萃取时间以及P204浓度对L-辛弗林萃取率的影响。 通过原子力显微镜观察了P204/正辛醇反胶束;萃取L-辛弗林的最佳条件为：萃取原料液水相pH=6.5,含水量W0=15、P204浓度为0.08 mol/L,在此条件下萃取15 min,对L-辛弗林的单次萃取率为68.0%。 基于萃取率建立的数学模型可反映pH值、P204浓度与萃取率的关系,在一定pH值及P204浓度区间可预测萃取率的变化趋势。     

苦参配方颗粒的高效液相色谱指纹图谱研究及定量分析

研究了苦参配方颗粒的指纹图谱和含量测定。采用高效液相色谱法,在C18柱上以乙腈-磷酸氢二钠水溶液(20 mmol/L,磷酸调pH 7.1)进行梯度洗脱,柱温35℃,检测波长220 nm,用指标成分氧化苦参碱、氧化槐果碱和苦参碱进行了定位,并在同一色谱条件下同时测定了它们的含量。确定了15个共有峰,其中7、8和11号峰分别为氧化苦参碱、氧化槐果碱和苦参碱。10批样品平均相似度为0.99以上,三个成分的含量分别为:氧化苦参碱50.0~79.8 mg/g;氧化槐果碱13.5~18.1 mg/g;苦参碱16.5~30.1 mg/g。结果表明:指纹图谱与指标成分含量测定相结合,可提高苦参配方颗粒的内在质量控制水平。     

微波辅助提取-高效液相色谱法测定苦参中5种苦参生物碱的含量

取粒径在0.18～0.25mm的药材粉末于提取罐中,用水作提取剂,在110℃、压力为15MPa、功率为780W的条件下进行微波提取(PMAE)5min,经离心后取上清液为提取液。提取液经无水乙醇处理提纯后得到待测物的甲醇溶液,从中分取部分溶液按选定条件用液相色谱-质谱(LC-MS)联用做定性分析,用高效液相色谱(HPLC)做定量分析。LC-MS结果表明:提取液中含有氧化苦参碱、氧化槐果碱、槐定碱、苦参碱和槐果碱,与其它3种提取方法比较,PMAE的提取率高且提取时间短。HPLC定量时,采用C18色谱柱,以不同比例的甲醇、乙腈和0.1%氨水溶液(pH 10.3)的混合溶液作流动相进行梯度淋洗,可分离上述5种生物碱,以峰面积和浓度间的线性关系进行定量。5种生物碱的回收率在93.9%～101.0%之间,测定值的相对标准偏差(n=5)在0.53%～2.6%之间。     

液相微萃取/后萃取-高效液相色谱法测定氧化苦参碱和苦参碱

建立了液相微萃取/后萃取-高效液相色谱法测定中药苦参、复方苦参注射液中氧化苦参碱和苦参碱含量的方法。利用自制的微萃取装置,选择异丙醇为萃取有机溶剂,2.00 mL NaOH(pH9)为供相,HCl(pH4)为接受相,聚丙烯腈纤维的长度为10 cm,搅拌速度为1500 r/min,萃取时间为30 min。萃取完成后,经高效液相色谱仪分析,测得氧化苦参碱和苦参碱线性范围分别为11~437 mg/L和10~433 mg/L;检出限均为1.0mg/L;相对标准偏差分别小于9.4%和6.7%。复方苦参注射液中氧化苦参碱和苦参碱的平均回收率分别为83.0%~116.1%和108.8%~117.8%;苦参药材中氧化苦参碱的平均回收率为104.3%~114.7%。本方法有机溶剂用量少,可有效去除复杂机体的干扰,测得结果满意。     

液相色谱/光电二极管阵列检测/质谱联用技术在中药分析中的应用

建立了基于液相色谱/光电二极管阵列检测/质谱(LC/DAD/MS)技术的中药分析基本方法。检出了丹参水溶性成分中的丹参素、原儿茶醛、咖啡酸、紫草酸、丹参酸丙和丹酚酸C等;在银杏内酯提取物中,鉴定和发现了白果内酯,银杏内酯A、B、C和一些新化合物,其中经结构确证的两个新化合物分别被命名为银杏内酯K和L;在苦参生物碱中鉴定了苦参碱、氧化苦参碱、槐果碱或异槐果碱、N-氧化槐果碱和槐花醇N-氧化物。     

电解质水溶液在丙酸十二铵-四氯化碳溶液中的增溶

表面活性剂在非极性溶剂中形成的反胶束在催化反应、光化学、蛋白质苹取分离等方面有着广泛的应用问．这些应用与反胶束的性质有着密切的关系，而增溶水后的反胶束其形状和大小都会发生很大的变化．增溶不同水量的反胶束的微极性、酸碱性、微勤度等已有不少文献报导［2－5］．一些不溶于非极性溶剂而溶于水的物质可以溶解在非极性溶剂中的反胶束核心水团中，这个现象被称为二次增溶．其中，电解质的二次增溶对于研究配体转换反应。酶催化反应问及改变反胶束内部的微环境有着十分重要的作用，Aebi和Weibush回首先研究了有水存在时N。CI在A…     

三烷基混合叔胺-正辛醇-磺化煤油萃取菜籽粕酸脱毒液中的植酸

用三烷基混合叔胺(N235)-正辛醇-磺化煤油从菜籽粕酸脱毒废液中萃取植酸,研究了水相pH值、植酸浓度、N235浓度对植酸分配比的影响。 确定萃取植酸的最佳条件为：初始pH=3.0、N235的浓度0.17 mol/L、正辛醇浓度0.48 mol/L、相比R为5∶1、萃取时间5 min,对0.02 mol/L标准植酸溶液的萃取率为95.2%。 以0.25 mol/L NaOH溶液为反萃取剂,在相比5∶2下反萃取5 min,三级反萃取总收率达到82.6%。 实验证明加入正辛醇可提高N235萃取植酸的分配比,可能与正辛醇和N235形成“分子簇”结构产生协同萃取有关。     

以离子液体作流动相添加剂的高效液相色谱法测定复方苦参注射液中4种生物碱

将离子液体1-丁基-3-甲基咪唑四氟硼酸盐(［BMIM］BF4)作为流动相添加剂建立了同时测定复方苦参注射液中4种主要生物碱的HPLC分析方法。以Agilent TC-C18柱(250 mm×4.6 mm, 5 μm)为分离柱,甲醇-0.1%磷酸水溶液(含2.2×10-4mol/L ［BMIM］BF4)(5:95, v/v)为流动相,流速1.0 mL/min,柱温30 ℃,进样量20 μL,在205 nm下检测。结果表明,苦参碱、槐定碱、氧化槐果碱以及氧化苦参碱的质量浓度分别在25.8～155.0 mg/L, 40.0～240.0 mg/L, 21.7～130.0 mg/L和37.5～225.0 mg/L范围内线性关系良好,相关系数均大于0.9990,平均回收率(n=9)在96.2%和98.9%之间。离子液体的加入能明显改善C18柱分离生物碱的色谱峰形并提高分离度。本法简便、快速、重复性好,可用于复方苦参注射液中生物碱的分离与测定。     

液-质联用研究槐定碱、槐果碱和苦参碱在兔体内的药代动力学

建立了一种液-质联用测定家兔血浆中槐定碱、槐果碱和苦参碱浓度的方法,并用于生物碱的药代动力学研究。分析方法的线性范围为槐定碱13.2~995.0μg/L,槐果碱7.0~530.0μg/L和苦参碱8.8~655.0μg/L,回收率为90.2%~99.8%。药代动力学参数计算结果表明,槐定碱和槐果碱符合二室模型,而苦参碱则符合三室模型。比较苦黄注射液和混合标准品注射后的药代动力学参数,表明苦黄注射液中的其他成分对槐果碱有促进其消除的作用,对槐定碱的消除可能有促进作用,而对苦参碱则没有影响。     

Simultaneous determination of matrine, sophoridine and oxymatrine in Sophora flavescens Ait. by high performance liquid chromatography

Following a detailed study, a reversed-phase high performance liquid chromatographic method (HPLC) has been developed and validated for analysis of three bioactive alkaloids, matrine, sophoridine and oxymatrine, in Sophora flavescens Ait. HPLC separation of the alkaloids was performed on a Kromasil C(18) column and detected by ultraviolet absorbance at 208 nm. The column temperature was maintained at 40 degrees C. A mobile phase composed of 0.01 mol/L KH(2)PO(4) buffer-methanol-triethylamine in the ratios 94:6:0.01 (v/v) was found to be the most suitable for this separation at a fl ow-rate of 1.0 mL/min and enabled the baseline separation of the three analytes free from interferences with isocratic elution. The analysis time was 24 min per injection. The calibration was linear in the range of 0.2-120.0 micro g/mL for matrine, 0.2-115.2 micro g/mL for sophoridine and 0.2-110.4 micro g/mL for oxymatrine, respectively. For assaying Sophora Flavescens Ait. samples, the relative standard deviations were 2.0% for matrine, 2.8% for sophoridine and 1.8% for oxymatrine analysis. The average recoveries of matrine, sophoridine and oxymatrine were 93.9, 95.3 and 93.5% for the Sophora flavescens Ait. samples, respectively. The method has been successfully applied to the simultaneous determination of matrine, sophoridine and oxymatrine in Sophora Flavescens Ait. samples collected in different habitats.     

Extraction of Bovine Serum Albumin Using Reverse Micelles Formed by Hexadecyl Trimethyl Ammonium Chloride

The extraction of bovine serum albumin (BSA) has been investigated using reverse micelles of hexadecyl trimethyl ammonium chloride/n-octanol/isooctane. Forward extraction process parameters such as the surfactant concentration, co-solvent concentration, pH, ionic strength, and species of the initial aqueous phase were important factors affecting the extraction performance. These parameters were varied to optimize the extraction efficiency. Under the optimized conditions, forward extraction efficiencies of BSA can reach practically 99.55%. The thermodynamic study revealed that the extraction of BSA is controlled by entropy changes. Maximum back-extraction efficiency of 85.16% can be obtained at low pH values and high salt concentrations. The structures of BSA during reverse micelle extraction did not change by comparing the circular dichroism spectra of BSA back-extracted to the aqueous phase with that of feed BSA.     

Field-amplified sample stacking in capillary electrophoresis for on-column concentration of alkaloids in Sophora flavescens Ait

A simple and rapid capillary zone electrophoresis method was developed for the separation of the main alkaloids from Sophora flavescens Ait. with the optimum buffer solution containing 110 mM NaH(2)PO(4) and 15% 2-propanol (pH 3.0). The field-amplified sample stacking (FASS) technique was applied to the on-line concentration of the alkaloids. The data presented in this work demonstrate that the use of a short water plug at the column inlet is essential for improving the reproducibility of FASS with electro-injection, and that the water plug injection time affected the sensitivity significantly. The sample concentration was further increased by about 2-3-fold by the introduction of a relatively longer water plug. With this stacking measure, the concentration sensitivity was about 3-4 orders of magnitude higher than in hydrodynamic injection.     

AEOT反胶束中脂肪酶的催化活性

反胶束已广泛应用于膜模拟化学和蛋白质的液 液萃取中[1～ 3] ,反胶束酶反应作为实现有机相酶催化的方法之一 ,具有许多独特的优点 ,反胶束独特的结构特征使表面活性剂分子组成的膜将油水相隔开 ,从而有利于保持酶的活性和稳定性。酶在反胶束的微水环境中比在水溶液中更接近天然的细胞内环境 ,在这里酶和底物分子均可得到有效的分散 ,接触几率大大提高 ,因而催化效率也得到很大提高。反胶束可以适用于各种类型的 (亲水的、疏水的和双亲的 )底物[4] ,已逐步形成“胶束酶学”的研究分支 ,研究胶束酶学的Martinek等[3] 曾预言 :反胶束体系有可…     

Reverse Micellar Extraction of β-Galactosidase from Barley (Hordeum vulgare)

The reverse micellar system of sodium bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane was used for the extraction and primary purification of beta-galactosidase (EC 3.2.1.23) from the aqueous extract of barley (Hordeum vulgare) for the first time. The process parameters such as the concentration of the surfactant, the volume of the sample injected, and its protein concentration, pH, and ionic strength of the initial aqueous phase for forward extraction, buffer pH, and salt concentration for back extraction are varied to optimize the extraction efficiency. Studies carried out with both phase transfer and injection mode of reverse micellar extraction confirmed the injection mode to be more suitable for beta-galactosidase extraction. The extent of reverse micellar solubilization of proteins increased with an increase in protein concentration of the feed sample. However, back extraction efficiency remained almost constant (13-14.4%), which indicates the selectivity of AOT reverse micelles for a particular protein under given experimental conditions. beta-Galactosidase was extracted with an activity recovery of 98.74% and a degree of purification of 7.2-fold.     

Microwave-assisted extraction of oxymatrine from Sophora flavescens

In this paper, microwave-assisted extraction (MAE) of oxymatrine from Sophora flavescens were studied by HPLC-photodiode array detection. Effects of several experimental parameters, such as concentration of extraction solvent, ratio of liquid to material, microwave power, extraction temperature, and extraction time on the extraction efficiencies of oxymatrine were evaluated. The optimal extraction conditions were 60% ethanol, a 20:1 (v/v) ratio of liquid to material and extraction for 10 min at 50 °C under 500 W microwave irradiation. Under the optimum conditions, the yield of oxymatrine was 14.37 mg/g. The crude extract obtained could be used as either a component of some complex traditional medicines or for further isolation and purification of bioactive compounds. The results, which indicated that MAE is a very useful tool for the extraction of important phytochemicals from plant materials, should prove helpful for the full utilization of Sophora flavescens.     

用磷酸二异辛酯/正辛醇反胶束从面包酵母粗提液中萃取谷胱甘肽

采用磷酸二异辛酯（P204）/正辛醇反胶束液从面包酵母粗提液中萃取还原型谷胱甘肽(GSH),研究了各种因素对萃取率的影响以及水相pH值和P204浓度与GSH萃取率关系的数学模型。正交实验确定,GSH的最佳萃取条件是水相pH值2.0,K+浓度0.1 mol•L-1,萃取时间15 min,P204浓度0.16 mol•L-1,GSH单次的萃取率为33.1%,2次萃取率达到49.7%。