肠癌患者尿中核苷排放的高效液相法研究

 用反相高效液相法测定尿中核苷。通过苯基硼酸亲和法提取尿中核苷,在柱(4 6mmi d ×250mm,5μm)上以25mmol/L磷酸二氢钾溶液(pH4 55)和60%的甲醇水溶液作为流动相进行二元梯度淋洗,于22℃下进行反相分离,260nm处紫外检测。用该法测定了41例肠癌患者和52例正常人尿中15种核苷的含量(用核苷与肌酐的摩尔比表示,下同),结果表明肠癌患者中有12种核苷的含量比正常人显著性增高(P<0 001)。以15种核苷的含量作为参量,结合主成分分析区分正常人和肠癌患者,对癌症病人的识别率达76%(31/41)。     

High Performance Liquid Chromatography of Dithiolthiones

Abstract

Sixteen dithiolthiones including oltipraz (5-(2-Pyrazinyl)-4-methyl-1-1, 2 dithiol-3-thione) and anethol dithiolthione (P-methoxyphenyl-1,2-dithiol-3-thione) have been studied using an isocratic high performance liquid chromatographic system. The retention characteristics of these compounds were determined using a conventional C 18 reverse phase column and a μ Bondapack phenyl column. The mobile phases used consisted of several concentrations of methanol and water. All compounds studied were adequately detected in the nanogram range (30–100 ng on column) using ultra violet detection (UV) set at 300 nm. Several analogs and isomers were separated. The retention characteristics of the 16 compounds studied are reported for both columns using 4 mobile phases and chromatograms of reference standards are presented.     

Reversed-Phase High Performance Liquid Chromatography (HPLC) of Nucleosides in Cell Extracts with Special Reference to Deoxythymidine

Abstract

A reversed-phase high performance liquid chromatography (HPLC) system was designed to assay the nucleoside concentrations, especially deoxythymidine, of tissue cultured cells. Concentration and purification of the acid extracted cell samples were achieved by utilizing a Sep-Pak C₁₈ cartridge. After adsorption of the sample, the cartridge was washed with 0.5 ml of water followed by 2 × 0.5 ml of 2.5% methanol. The compounds of interest were subsequently eluted in 2 × 0.5 ml of methanol. The cartridge procedure was found to be fast, inexpensive and showed good recoveries for most tested nucleosides. The nucleosides uridine, deoxythymidine and adenosine could be detected in green monkey kidney cells. Ribonucleotides and deoxyribonucleotides could be separated from each other with the HPLC system used.     

Fluorescence Detection in High Performance Liquid Chromatography

Abstract

The principles and applications of fluorescence detection and fluorescence introducing reagents and methods in HPLC are reviewed. The design and requirements for fluorescence detectors, flow cells and excitation sources and the conversion of non-fluorescent compounds into fluorescent products by pre-column and post-column derivatization reactions are discussed. For the applications the emphasis is on drug analysis, where possible in biological fluids (serum, urine, etc.). The last paragraphs are divided in a number of sections in which newly developed and some scarcely used reagents are mentioned shortly; a more complete treatment is given of the reagents and labels most frequently used in the derivatization of certain functional groups. In this discussion the methods of derivatization as well as the selectivity, stability, fluorescence behaviour of the reagents/labels and derivatives and the reaction conditions are included. An up-to-date survey of the applications of fluorescence detection in liquid chromatography (TABLE III, TABLE IV and TABLE V), ends this review paper.     

气相色谱法和高效液相色谱法分析中不确定度的评定

气相色谱法和高效液相色谱法在标准曲线的绘制、标准样品的使用、测定结果的输出等方面极为相似,因而,评定其分析不确定度时具有相同的数学模型,且各不确定度分项的评定可以采用相似的评定方法。本文参照有关文献[15],以高效液相色谱法测定胺苯磺隆为例,对其不确定度的评定进行了讨论。1　主要仪器、试剂和实验方法　　HPLC1100型高效液相色谱仪,进样定量环(20μL,不确定度1%);电子分析天平。容量瓶:A级,100×(1±2×10-3)mL,50×(1±2×10-3)mL;刻度移液管:分度值0 01mL;胺苯磺隆标样:质量分数w为99 0%,标准偏差S为0 01%(n=5)。　…     

超声波破碎-高效液相色谱法定量检测核酸

采用超声波破碎结合高效液相色谱技术,建立了定量检测质粒DNA的方法,测量结果可以溯源至核苷酸标准物质.采用超声波破碎(功率300 W,频率24 kHz)技术将质粒DNA破碎成200～500 bp的小片段DNA,再用蛇毒磷酸二酯酶将其水解为4种核苷酸(dCMP:3.2 min;dTMP:4.7 min; dGMP:5.3...     

High Performance Liquid Chromatography of Uroporphyrin Isomers

Abstract

The separation of uroporphyrin I and III isomers by reversed-phase high performance liquid chromatography on ODS-Hypersil with ammonium acetate buffer-acetonitrile solvent systems is described. The effects of buffer concentration, pH, organic modifier proportion and different organic modifiers on the resolution are studied. The optimum conditions for the separation were 12–13% acetonitrile in 1M ammonium acetate buffer pH 5.10–5.20. The method also separated uroporphyrin I and III from the II isomers but the resolution of uroporphyrin III and IV isomers was not achieved.     

High Performance Liquid Chromatography of Cholesteryl Esters

Abstract

The separation of cholesteryl esters by high performance liquid chromatography was facilitated by cooling of the column, followed by spontaneous return to ambient conditions. the resolution of the esters was increased. the method was applied to detection of the cholesteryl esters in plasma.     

蛋白质制备色谱的研究

研究了蛋白质纯化制备的二种运行模式,其一,在色谱柱超载下纯化制备蛋白质模式,并成功地用离子交换色谱纯化制备了大豆中的胰蛋白酶,用反相液相色谱纯化了细胞色素Ｃ;其二,用溶质－溶质顶替色谱纯化制备了核糖核酸酶－Ａ,并对顶替色谱过程中诸参数进行了讨论和选择。     

分子印迹液相色谱整体柱

分子印迹是合成预定选择性固定相的新兴技术,整体柱是新型的色谱固定相技术。将分子印迹聚合物与整体柱技术相结合,可以有效提高液相色谱的分离效率,有助于推动整个分离科学的发展,意义重大,是当今分析化学的研究热点。本文就分子印迹液相色谱整体柱的制备合成、色谱分离条件以及物理化学特性评价方法等方面的研究进展进行了较系统的综述,并对该技术目前存在的问题和发展前景进行了探讨。     

Agilent 1100高效液相色谱系统

色谱分析是目前最为活跃的分析化学分支 ,在环境、生化、药物分析等领域的应用很普遍。高效液相色谱法分离效能高、选择性好、检测灵敏度高、分析速度快、仪器稳定性和数据的重现性均较好。近年来 ,随着高效液相色谱仪器的发展及完善 ,特别是计算机技术的引入 ,可以实现分析条件的快速优化选择和准确的定量处理 ,解决实际问题的能力日益增强 ,应用范围不断扩大 ,已经成为很多实验室必备的分离分析和制备纯化手段。国内外有多家仪器制造商可提供高效液相色谱仪 ,其中安捷伦科技公司 (原惠普公司的分析仪器部 )生产的高效液相色谱仪经多年的发…     

Hypnotic-Sedative Screen by High Performance Liquid Chromatography

Abstract

In this procedure some of the most commonly abused sedatives and hypnotics can be identified and quantitated using 100 ul of serum. Ethosuximide, primidone, methyprylon, phenobarbital, butabarbital, butalbital, glutethimide, mephobarbital, phenytoin, carbamazepine, secobarbital, phensuximide, pentobarbital, amobarbital, and methaqualone are extracted into a chloroform-ethanol solvent containing either 5-(p-methylphenyl)-5-phenyl-hydantoin or hexobarbital as an internal standard, evaporated to dryness, reconstituted with mobile phase and injected into a high performance liquid chromatograph in conjunction with a radial compression separation system. Peak heights are measured at 195 nm and 254 nm and sensitivity for all drugs is one μg/ml. Day to day precision obtained CV's ranging from 4.5 to 10.4%.     

高效液相色谱整体柱技术的进展

高效液相色谱整体柱（又名连续床）具有制备相对简单、原料易得以及聚合组分在一定范围内可调节的优点,是近年来得到迅速发展的新型色谱柱。本文综述了目前高效液相色谱（HPLC）制备整体柱的典型高聚物体系、制备各种整体柱时反应条件的影响,并简要介绍了它的表征方法和应用。     

High Performance Liquid Chromatography of Zinc and Copper Pheophytins

Abstract

The zinc or copper chelates of pheophytins a and b were formed and separated on a reversed-phase C-18 column. Allomerized products were produced readily during the chelation reaction. Resolution of the allomerized compounds from the non-allomerized chelates was achieved using a gradient elution technique. Compound identification was facilitated by monitoring the column eluate at both 436 and 658 nm. The method allowed for isolation of individual pigments for further study.     

高效液相法同时检测南昌霉素和梅岭霉素

 采用高效液相法同时分离测定聚醚类抗生素南昌霉素和十六元大环内酯类抗生素梅岭霉素。柱为WatersXTerraTM RP18柱 (3 9mmi.d .× 15 0mm ,5 μm) ,流动相为乙腈水溶液 ,流速为 0 8mL/min ,其梯度洗脱程序为 5 7% (体积分数 ,下同 )乙腈 (30min)→ 70 %乙腈 (2min)→ 80 %乙腈 (2min)→ 90 %乙腈 (16min)。检测波长为 2 34nm ,柱温 2 5℃。该法测定的灵敏度好 ,加标回收率高 ,可用于南昌链霉菌发酵液中南昌霉素和梅岭霉素的定量测定。此法具有灵敏、准确、快速的特点。     

高效液相色谱法测定克芜踪

1 引言克芜踪(gramoxone)中的有效成分为百草枯有效离子,属于双吡啶盐类,为广谱触杀性除草剂.目前国内多采用比色法和气相色谱法测定百草枯的含量,不仅分析时间长,且回收率低.本实验采用高效液相色谱法,该方法简便、快速、准确,且回收率高,已应用于实际的进口克芜踪检验工作中.2 实验部分2.1 仪器和试剂 HP1090Ⅱ液相色谱仪(HP公司),配有二极阵列检测器(DAD);三元溶液系统;定量进样器及化学工作站;Millipore超纯水器.乙腈:分析纯,过滤;水:二次蒸馏水;百草枯标准品:纯度为99.6%;B-5离子对试剂(I-pentane sulfonic acid);H_2SO_4 (1:3);三乙胺:分析纯;滤膜:0.45μm.     

炔敌稗的高效液相色谱分析

炔敌稗(proyzamide),其化学名称是3,5-二氯-N-(1,1-二甲基丙炔基)苯甲酰胺,是一种应用广泛的除草剂,常用于阔叶杂草(wild oats,john grass,ryegrass)等杂草的控制.目前,只有用高效液相色谱法检测其在环境中残留量的报道,但该方法所用试剂昂贵,操作繁琐,不适用于产品的质量分析,本文建立了测定炔敌稗有效成分的高效液相色谱定量分析方法,并利用质谱(MS)、核磁共振(NMR)确定其中的主要杂质为3-氯-N-(1,1.二甲基丙炔基)苯甲酰胺.该方法可用于产品的质量检测.2 实验部分2.1 仪器与试剂 P200型高效液相色谱系统,可变波长紫外检测器,WDL-95色谱工作站,样品过滤器(国家色谱分析中心,大连依利特科学仪器有限公司),数据用WDL-95色谱工作站在联想P5/100微机上处理.Bruk-er Ac-P200核磁共振仪.HP5989质谱仪.炔敌稗粉剂(50%)为Rohm-Haas公司产品;水为二次蒸馏水;甲醇为色谱纯;炔敌稗标准(自制).     

High Performance Liquid Chromatography of Tryptophan and Serotonin Metabolites

Abstract

Serotonin is a neurotransmitter in cerebral centers and its perturbations can produce humor and behavior disorders. In addition, exploration of tryptophan and serotonin metabolism is extremely important for early detection and supervision of treatment in carcinoïd tumors. High performance liquid chromatography (HPLC) enabled us to separate and titrate different metabolites (5-hydroxyindolylacetic acid, serotonin, tryptophan, 5-hydroxy-tryptophan and N-acetyl tryptophan (NAT) as internal standard) in several types of biological tissues: blood, urine, brain and cerebrospinal fluid. This method is original because it connects HPLC with fluorescence in continuous flow, which allows to change the conditions of pH and buffer. This technique is highly sensitive (below 100 picograms) and very quick (ten minutes).     

Ion Exchange High Performance Liquid Chromatography of Leukotrienes

Abstract

A novel anion exchange liquid chromatographic system has been developed for isocratic separation of leukotrienes. Hydrophobic as well as ionic forces were found to influence the separation. By optimization of solvent strenght, ionic strenght and pH, amphoteric peptidoleukotrienes could be separated simultaneously with hydroxy fatty acids such as leukotriene B₄ and its ω-oxidized metabolites. To obtain a good buffering capacity of the mobile phase at optimum pH, a multicomponent buffer was developed.     

高效液相色谱法测定氟喹诺酮类药物

1 引 言氟喹诺酮类药物(fluoroquinolone,FQS)由于具有抗菌谱广、抗菌作用强、生物利用度高、毒副作用小、组织分布广等特点,而在临床上得到广泛应用。有关高效液相色谱法测定FQS的方法已有较多文献报道,但由于FQS结构上的相似性,多限于单组分的测定或多组分梯度洗脱分离。而采用同一流动相同时测定多种FQS却少见报道。本文建立了一种简