高效液相色谱法分离测定香兰素

用６０％乙醇提取香荚兰豆的香兰素,经高效液相色谱法分离,在紫外波长３３０ｍｍ处测定,选取适当的流动相,能快速,准确地测定出香兰素,最低检测含量可达到０．０１μｇ．     

反相高效液相色谱法制备HMC毒素纯品

本文采用反相高效液相色谱纯化制备了从玉米小斑病病菌Ｃ小种（ＨｅｌｍｉｎｔｈｏｓｐｒｉｕｍｍａｙｄｉｓＲａｃｅＣ）分离出来的毒素ＴｏｘｉｎＩ,经ＮＭＲ的结构分析证实此制备物纯度较高,达到结构分析要求。用氯仿提取含有ＨＭＣ的毒素培养滤液,经过多次ＴＬＣ展层层析（ＣＨＣｌ_３：ＭｅＯＨ＝９：１）,刮取具有侵染活性的部分（Ｒ_ｆ＝０．４）溶解在小体积甲醇中备用。色谱柱为Ｗａｔｅｒｓ（制备型２５０ｍｍ×１０ｍｍ）填料为ＹＷＧ－Ｃ１８反相柱,粒度１０ｕｍ,流动相选用甲醇：水５５：４５     

制备液相色谱技术

液相色谱技术的发明,可以追溯到本世纪初。俄国植物学家茨维特以填装碳酸钙粉末的玻璃柱,分离植物色素的经典实验。自50年代之后,气相色谱、液相色谱等迅速发展,各种型式的色谱方法,成了分离和分析的利器,在科学技术的众多领域中发挥着重要作用。自70年代以来,高效液相色谱得到了飞速发展。     

高效液相色谱法分离测定香兰素

用60％乙醇提取香荚兰豆的香兰素,经高效液相色谱法分离,在紫外波长330nm处测定。选取适当的流动相,能快速、准确地测定出香兰素,最低检测含量可达到0．01μg。     

高效液相色谱法分离和鉴定腐马素B_1

腐马素是由串珠镰刀菌产生的真菌毒素,其中腐马素B1可引发马脑白质软化症等疾病。本文用高效液相色谱法从串珠镰刀菌玉米培养物中分离出了腐马素B1并通过紫外光谱和快原子轰击质谱进行了鉴定。     

高效液相色谱法测定辣椒素

用９５％醋酸钠饱和乙醇溶液抽提辣椒树脂油中的辣椒素,经高效液相色谱分离,在紫外长２８０ｎｍ处测定。用此法分析测得福建特产“小米椒”的辣椒素含量为１０．３８％。     

高效液相色谱法分离和鉴定腐马素B1

腐马素是由串珠镰刀菌产生的真菌毒素,其中腐马素Ｂ１可引发马脑白质软化症等疾病。本文用高效液相色谱法从忠珠镰刀菌玉米培养物中分离出了腐马素Ｂ１并通过紫外光谱和快原子轰击质谱进行了鉴定。     

制备色谱操作条件的优化

色谱优化是蛋白质生产制备工艺开发中必不可少的一步。本文讨论了不同的优化目标 ,介绍了目前常见的优化方法 ,特别是实际应用中的原则及实验方法 ,对涉及优化的基本概念也做了说明。     

Separation of Glycopeptides by High Performance Liquid Chromatography

A method is presented for separation of tryptic glycopeptides-containing oligosaccharides of the N-asparagine-linked type. High performance liquid Chromatography (HPLC) of glycopeptides on a C18 reverse-phase system eluted with a gradient of 0%–50% acetonitrile in 0.1 M NaPO₄ pH 2.2 resolves the two major glycosylation sites from the envelope glycoprotein (G) of vesicular stomatitis virus. Glycopeptides containing N-linked oligosaccharides of the complex type coelute with those containing N-linked oligosaccharides of the neutral, high mannose type, indicating that separation is based upon peptide rather than carbohydrate composition. The contribution of the carbohydrate component to glycopeptide elution, as determined by cleavage of the high mannose oligosaccharides with endo-β-Nacetylglucosaminidase H, is that of a significant, but minor, decrease in peptide retention time. Comparison of the tryptic glycopeptide profiles of G isolated from both wild type and mutant strains of VSV illustrates the rapid, reproducible, and quantitative nature of the technique. Through HPLC analysis of appropriately treated glycopeptides, it is possible to explore both the nature and extent of glycosylation at individual sites in glycoproteins in a single step.     

高效液相色谱法测定辣椒素

用95%醋酸钠饱和乙醇溶液抽提辣椒树脂油中的辣椒素,经高效液相色谱(HPLC)分离,在紫外波长280 nm处测定。用此法分析测得福建特产"小米椒"的辣椒素含量为10.38%。     

双水相电泳分离蛋白质的研究

近几年来,随着生物技术的迅速发展,制备型电泳技术的研究得到了重视。然而由于技术上的原因,大规模的制备型电泳技术的研究还未能取得突破。阻碍电泳放大的一个主要问题是由于电加热作用而导致的热对流对电泳分离的破坏。为解决这一问题,人们提出了许多方法。例如,在太空的微重力环境下进行电泳,应力稳定自由流动电泳,循环等电聚焦和区带电泳,色谱电泳和等电膜等电聚焦等。这些方法在电泳放大上都取得了一定的进展,但各有其局限性。最近,Clark提出利用双水相的液液界面阻止热对流的设想,为开发大规模的制备型电泳技术开辟了一条新途径、Raghava Rao等在两种双水相体系上施加电场后成倍地缩短了分相时间。Levine和Bier采用U型管电泳装置研究了双水相体系中血红蛋白的电泳迁移率,观测到界面有阻滞作用。Clark在柱型电泳装置中进行了一组双水相萃取肌红蛋白的简单实验。在10mA的恒电流下电泳40min之后,肌红蛋白的分配系数为7.5,而当电场反向后,分配系数变为0.04,界面阻力并不显著,两者结论并不一致。     

A Quantitative Regional Analysis of Amino Acids Involved in Rat Brain Protein Synthesis by High Performance Liquid Chromatography

A biochemical method is described for the simultaneous quantitative estimation of unidirectional blood-brain amino acid influx and protein biosynthesis in individual structures of the rat brain. The method involved a double labeling experiment started by the administration of [14C]carboxyl-labeled amino acids and terminated 2 min after infusion of 3H-labeled amino acids, each at tracer quantities, the total labeling period being 45 min. Specific radioactivities of 14C- or 3H-labeled phenylalanine, tyrosine, leucine, isoleucine, and valine were determined in plasma and in small brain tissue samples for free amino acids, aminoacyl-tRNAs, and proteins. Amino acids were converted to their corresponding 5-dimethylamino-naphthalenesulfonyl (Dns, dansyl) derivatives and separated on HPLC C18 reversed-phase columns isocratically according to a newly developed optimizing procedure. The order of influx values between the neutral amino acids in relation to each other was Leu greater than Tyr greater than Ile greater than Phe greater than Val in every structure examined. Although aminoacylation of tRNAs was found to proceed to a comparable degree for neutral amino acids in all regions investigated, the specific radioactivity of amino acids attached to tRNAs differed substantially from that in the free amino acid pool, especially for leucine and valine. The results indicate the necessity of aminoacyl-tRNA determinations for tracer incorporation studies in protein synthesis analysis. Relative protein synthesis rates in the halothane-anesthetized rat were determined to be 30 and 67-91 pmol total amino acid incorporation/min/mg tissue for white and gray matter, respectively.     

高效液相色谱法测定血浆中同型半胱氨酸

建立了一种高效液相色谱-库仑阵列电化学测定血浆中同型半胱氨酸的方法。采用3-2-羧乙基膦作为还原剂还原巯基间形成的二硫键,用0.1 mol.L-1HC lO4来沉淀血浆蛋白,离心取上清,进样分析。在0.40~50.00μmol.L-1范围内,线性关系良好。加标平均回收率是98.43%,RSD%是4.35%。该方法简单、快速、灵敏。     

Preparation of Porcine Neurophysin Proteins by High Performance Liquid Chromatography

Abstract: The crude neurophysin containing extract from posterior lobes of porcine pituitaries was roughly purified by gel chromatography. 15 mg of the lyophilized neurophysin complex were completely separated by HPLC yielding in neurophysin I₁ (3.6 mg), I₂ (4.0 mg), II (4.6 mg) and III (1.9 mg). All of the neurophysins were homogenous by PAGE and SDS-electrophoresis, isoelectrofocussing, amino-acid composition and N- and C-terminal amino acid analysis. In conclusion, HPLC is a reliable and quick method for the preparation of pure neurophysins.     

高效液相色谱测定血清中的非胆固醇甾醇

本文介绍一种用高效波相色谱（ＨＰＬＣ）同时测定血清中２４－脱氢胆固醇、７－烯胆甾烷醇、菜油甾醇、胆甾烷醇和β－谷甾醇的方法。血清加内标（６－氯豆甾醇）后用氢氧化钾醇溶液皂化,用正己烷提取其中的各种甾醇,将提出的甾醇衍生为苯氨基甲酸酯,用ＨＰＬＣ分离测定。本法样品处理简单,色谱分析时间短,批内和批间变异系数分别为２．２～３．８％和３．３～６．７％。本文也首次报告我国成年人血清非胆固醇甾醇水平,５种甾醇血清浓度的总和平均为１．５４ｍｇ／ｄｌ,是胆固醇的０．８６％。     

高效液相色谱测定微量胆固醇氧化产物

介绍一种高效液相色谱测定胆固醇氧化产物的方法,以苯甲酰氯为衍生剂,将胆固醇及其氧化产物衍生成苯甲酸酯,反相高效液相色谱分离,紫外检测,内标法定量．本法灵敏度高,重现性好,可应用于胆固醇纯度标准物质的杂质分析及血清中的胆固醇氧化产物测定。     

中压制备液相色谱快速分离制备儿茶素单体

以反相中压制备液相色谱为工具,甲醇-水作为洗脱溶剂,通过C18 (ODS-AQ)填料从茶多酚中一步分离出表没食子儿茶素(EGC)、表没食子儿茶素没食子酸酯(EGCG)、表儿茶素(EC)、表儿茶素没食子酸酯(ECG)等四种儿茶素单体.1.0g纯度为92.6％的茶多酚经过中压色谱,制备得到了90 mg EGC、355 mg EGCG、23 mgEC和92mg ECG,它们的纯度分别为91.8％、97.6％、97.7％、99.3％,纯品得率56.0％,四种单体总回收率达到68.2％.四种儿茶素单体的结构经核磁共振氢谱、碳谱以及高分辨质谱加以确证.