A simplified method for isocratic HPLC analysis of polyamines

A simple technique is described for the separation and analysis of polyamines in tissues and body fluids, utilizing precolumn clean-up on disposable CM-cellulose columns, followed by an automatable HPLC procedure. Complete separation and analysis takes 12-15 min per sample with sensitivity in the pmole range.     

Development and evaluation of a method of high pressure liquid chromatography for the determination of the level of degradation of thyroliberin (TRH) in blood]

The degradation of the TRH by plasma was studied using a sensitive method for the separation of TRH and products formed by high-pressure liquid chromatographic analysis (HPLC). The detected products are TRH-3H or synthetic TRH. The HPLC analysis is performed on a microparticulate (10 mu) silica gel chromatographic column with CH3CN/0,01 M NH4OAc, at pH 6 (30/70) (V/V). This technique provides a good separation of TRH and the other products of degradation. In our experimental conditions, the human plasma degrades 62.5 +/- 5% of of TRH in 60 mn; very similar results are obtained with thin layer chromatography.     

Dünnschichtchromatographischer Nachweis der Aminosäuren im Urin

Summary Urinary amino acids are converted into DNP-derivatives. Extraction by organic solvents yields both ether- and acid-soluble DNP-derivatives in a form suitable for separation and identification by thin-layer chromatography. Solvent systems and techniques for the detection of 35 urinary constituents are described. The method is applicable to the qualitative analysis of other biological fluids.     

微纳流体样品片上分离技术

基于微流控技术的细胞或颗粒分离是样品前处理中的一个重要步骤,近些年在生物和化学分析中引起了极大关注,随着微纳米加工技术的发展,分离技术在片上实验室系统的微环境应用中得到快速发展.基于微流体细胞或颗粒的分选技术具有很多优点,包括减少了样品量、加速了样品处理速度、高灵敏度、高空间分辨率、成本低以及便携性等.本文介绍了当前最先进的基于连续微流体细胞或颗粒分选技术,从被动式与主动式分离两方面的典型实例详细阐述了各种分选原理及其在细胞或微颗粒分选中的应用,讨论了这些技术的主要指标,如分选标记、分辨率、效率以及处理量,同时讨论了各类在微流体下的微分离方法存在的缺点.微尺度下高效的微分离技术在实现片上实验室、现场微分析设备和家庭诊断中将会起着越来越重要的作用,引起了众多学者们的研究兴趣,能够促进微分离与检测和信号读取等集成于一体,形成一个便携式的诊疗设备.     

Resolution of small G1 and G2 populations of L1210 leukemia by flow microfluorometric analysis aided by velocity sedimentation

Summary L1210 leukemic cells grown in vitro were subjected to kinetic analysis using a flow microfluorometer. A single broad peak was found for the DNA content distribution if unfractionated cells were used; prior fractionation using lg velocity sedimentation allowed the separation of small peaks with smaller (G₁) and larger (G₂) DNA contents from the dominant S phase peak with intermediate DNA content.This work was supported by grant number 5P01CA13053 awarded by the National Cancer Institute, DHEW USA, and by grant num ber RBI 76-1 from the Queen Wilhelmina Fund, The Netherlands.     

Einige neue Trennungsmethoden und ihre Anwendung auf biochemische und organisch-chemische Probleme

Summary A survey is given of some recent progress, particularly in the author's own laboratory, of methods for the separation of macromolecules and particles of biological origin, as viruses, bacteriophage, microsomes, cell fragments, bacteria and whole cells. Special attention is given to recent developments in zone electrophoresis, protein chromatography, gel filtration and the new partition methods for separation of particles and macromolecules (Albertsson). A number of examples are given to illustrate the various applications. Special emphasis is laid upon the new possibilities for biochemical-genetic studies by application of high-resolution separation methods, and the significance of particle separation methods as applied to fragments of biological structures in providing a tool for structure investigations on a macromolecular level—a field which also should be of considerable interest from a chemical point of view.

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DrittePaul-Karrer-Vorlesung gehalten an der Universität Zürich am 5. Juli 1961.     

Rapid temperature programmed gas-liquid chromatography of volatile fatty acids (C1−C7) for the identification of anaerobic bacteria

Summary A gas liquid chromatography method for the separation of 10 volatile fatty acids (C₁–C₇ and isomers) has been improved by using oven temperature programmed conditions. In our conditions, the proprietary stationary phase SP 1220 introduced by Supelco Inc., gave sharp separation of volatile fatty acids in less than 8 min. This method was suitable for analyses with both thermal conductivity and flame ionization detectors.     

Resolution of small G1 and G2 populations of L1210 leukemia by flow microfluorometric analysis aided by velocity sedimentation

L1210 leukemic cells grown in vitro were subjected to kinetic analysis using a flow microfluorometer. A single broad peak was found for the DNA content distribution if unfractionated cells were used; prior fractionation using lg velocity sedimentation allowed the separation of small peaks with smaller (G1) and larger (G2) DNA contents from the dominant S phase peak with intermediate DNA content.     

A rapid HPLC method for determination of adenylate energy charge

A simple procedure is described for separation and analysis of adenine nucleotides in tissue extracts, utilizing anion exchange HPLC. Determination of AMP, ADP, and ATP takes 10 min per sample.     

动物外周血单个核细胞分离方法探讨

目的 探索用人淋巴细胞分离液分离多种属动物外用血单个核细胞（PBMCs）的可行性及评价其分离效果,建立一种适于野外大规模多种属动物PBMCs的分离方法.方法 采用人淋巴细胞分离液分离来源于新疆的伊犁马、新疆驴、不同品种犬、新疆双峰驼、天山马鹿和来源于重庆的荣昌猪、中国荷斯坦奶牛、简阳大耳黑山羊外周血PBMCs.随机抽样检测分离的动物PBMCs细胞总数、细胞纯度和细胞活率.结果 用人淋巴细胞分离液成功分离上述8种动物PBMCs,每毫升动物外周血分离的PBMCs细胞总数为0.52×106～2.03×106个,纯度为67%～93%,细胞活率为92.5%-98.0%.结论 用人淋巴细胞分离液分离多种动物PB-MCs的方法宜在动物的病毒分子流行学调查中进一步推广.     

The application of the high-performance ion-exchange chromatography for the analysis of bovine milk proteins

Summary A high-performance ion-exchange chromatographic (HPIEC) method for the analysis of milk proteins is described. The technique provides an alternative to other separation methods. The resolution obtained for both the proteins of the casein group and bovine lactoserum is comparable with that of electrophoretic analysis on polyacrylamide gels and low-pressure chromatography.     

早期母子隔离应激对大鼠学习记忆能力的影响

目的探讨早期母子隔离应激对幼鼠学习记忆能力的影响。方法30只SD新生大鼠用于实验,实验随机分成3组,正常对照纽、母子隔离15分钟纽、母子隔离3小时纽。在生后40天,三组大鼠进行Morris水迷宫及记录海马脑片长时程增强电位（LTP）实验。结果Morris水迷宫测试,与正常组比较母子隔离3h组大鼠寻找平台需要的潜伏期延长（P〈0．05）,而母子隔离15分钟组大鼠寻找平台需要的潜伏期与正常组比较则无明显变化（P〉0．05）;海马脑片长时程增强电位提示：正常对照组条件刺激（CS）前后fEPSP斜率变化率为：64．7±22．9％;母子隔离3h组为：35．3±14．2％,低于正常组（P〈0．01）;母子隔离15min组为：66．3±29．7％,与正常组比较无显著性意义（P〉0．05）。结论幼鼠生后早期过度的应激导致大鼠的学习记忆能力减弱。     

The application of the high-performance ion-exchange chromatography for the analysis of bovine milk proteins

A high-performance ion-exchange chromatographic (HPIEC) method for the analysis of milk proteins is described. The technique provides an alternative to other separation methods. The resolution obtained for both the proteins of the casein group and bovine lactoserum is comparable with that of electrophoretic analysis on polyacrylamide gels and low-pressure chromatography.     

N-terminal acetylation in a third protein family of vertebrate alcohol dehydrogenase/retinal reductase found through a 'proteomics' approach in enzyme characterization

A recent finding of a novel class of retinol-active alcohol dehydrogenase (ADH) in frog prompted analysis of this activity in other vertebrate forms. Surprisingly, yet another and still more unrelated ADH was identified in chicken tissues. It was found to be a member of the aldo-keto reductase (AKR) enzyme family, not previously known as an ADH in vertebrates. Its terminal blocking group and the N-terminal segment, not assigned by protein and cDNA structure analysis, were determined by electrospray tandem mass spectrometry after protein isolation by two-dimensional gel electrophoresis. The N terminus is Acetyl-Ala- and the N-terminal segment contains two consecutive Asn residues. The results establish the new ADH enzyme of the AKR family and show the usefulness of combined gel separation and mass spectrometry in enzyme-characterization.     

A practicable variant of the ion exchange method for the radiometric estimation of ornithine decarboxylase activity

Summary A known ornithine decarboxylase assay working with ion exchang separation of [³H]ornithine and [³H]putrescine has been revised. The assay can be performed in disposable 1.5 ml vessels with a total of four pipetting steps. The separation of enzyme substrate and product, respectively, requires 3 h per 50 samples. The detection limit is about 50 pmoles [³H]putrescine formed.     

Nonspecific reaction of a thiol:Protein disulfide oxidoreductase with the disulfide bonds of insulin

Summary A thiol:protein disulfide oxidoreductase from bovine liver was isolated after separation from protein disulfide isomerase. The enzyme, after activation (reduction) with glutathione, was reacted with stoichiometric amounts of insulin and the sulfhydryl groups of the partially reduced hormone were labeled with iodo (l-¹⁴C)acetamide. After separation of the insulin chains, the radioactivity was found in both the peptides, with a ratio A-chain/B-chain equal to 2/1.     

Verkürzte Methode zur Isolierung radiochemisch reinen Chlorophylls

Summary Radiochemically pure chlorophylla andb have been separated from barley by a rapid two-step purification procedure. It is based on chromatographic separation of pigments on polyamide columns and by paper chromatography.     

Gallen und Gallensäuren Papierchromatographische Untersuchungen

Summary In cases of bile-scretion disturbances substitution-therapy can be performed using suitable animal biles equivalent to human bile. Comparative investigations have been made to establish which animal bile is in practice the most suitable for bile substitutiontherapy based on its bile acid components.A new paper partition chromatographic method has been applied for separation and identification of free bile acids. Ascending development was used on Schleicher & Schüll 2043b Mgl paper impregnated with 20 v/v% propylene glycol in chloroform. The xylene-methylethylketone 1:1 solvent system gave good separation. The bile acids can be detected by immersing the chromatograms in 20 w/v% SbCl₃ in chloroform followed by drying and heating for 5–10 min at 100–110°C. The spots show intense reddish-violet or blue fluorescence in filtered UV-light (see Table 1).These investigations have shown that human and ox biles are, in contrast to pig bile, physiologically related biological substances regarding their bile acid components.     

Méthode de séparation des diverses classes d'ARN par électrophorèse sur gel de polyacrylamide en plaques

Summary A method of electrophoresis in polyacrylamide gel slabs for the separation of different classes of RNA is described. A gel mixture of 2.5% acrylamide and 0.5% agarose allows an improved and rapid separation of RNA molecules. Comparison between electrophoresis and centrifugation on a linear density gradient is discussed.     

人工蜂群优化算法在复数盲源分离中的应用

针对复值信号的源数估计和有序分离等关键技术,提出一种基于人工蜂群优化的源数未知的复值盲源分离方法,该方法首先利用交叉互验技术来估算复数源信号的个数,然后通过人工蜂群算法优化峰度的绝对值来获得最佳分离向量,并实现了逐次恢复源信号的目的.仿真实验结果表明,该方法不仅能依峰度绝对值的降序实现服从任何分布源信号的盲分离,同时比其他方法具有更优越的估计性能.另外,提出一种基于峰度的欠定复盲源分离算法,该算法根据信号的统计特性构造了用于欠定混合情况下盲抽取向量的代价函数,然后通过人工蜂群算法优化其函数来获得最佳分离向量,通过多次分离来实现欠定复盲源分离的目的.通过对混合分布类型的复值源信号欠定盲分离仿真实验验证了该算法的有效性.