New analytical methods for epoxy resins, 2. Two-dimensional chromatography

In order to gain information on the functionality type distribution and molar mass of bisphenol A based epoxy resins, on-line coupled two-dimensional chromatography was used. In the first chromatographic dimension, liquid chromatography at the critical point of adsorption separated the samples with respect to functionality. The functionality fractions were automatically transferred into size exclusion chromatography as the second dimension, where the fractions were separated with respect to their molar mass distribution. As a result of the two-dimensional experiment, the samples were mapped in contour plots, providing quantitative information on the interconnected parameters functionality, molar mass, and branching.     

Fractionation of the triacylglycerols of lipase-modified butter oil

Triacylglycerols (TGs) of lipase-modified butter oil were separated into saturated, monoene, diene and triene fractions on ap-propylbenzene sulfonic acid solid-phase extraction column loaded with silver ions. Fatty acid analysis of the fractions showed that the amounts of saturated TGs (98.4 mol%) and monoene TGs (26.0 mol%) in the saturated and monoene fractions, respectively, were close to the theoretical amounts of TGs in pure fractions. The column method provides a useful alternative to AgNO₃-thin-layer chromatography as a means of separating the TGs of butterfat and producing relatively pure TG fractions for further analysis by gas chromatography (GC) or GC-mass spectrometry.     

Molecular characterization of hydroxyl-terminated polybutadienes

A comparative study has been made on the functionality distribution of two hydroxyl terminated polybutadiene products which are used largely as prepolymers for sealants, adhesives, and solid propellants. They are known to possess nearly the same molecular characteristics. To this end, each product was fractionated into 17 fractions with different molecular weights by the preparative GPC technique. Sample fractions thus obtained were subjected to “polarity-controlled adsorption” thin-layer chromatography (TLC) to determine the functionality distribution. From these experimental results it was concluded that a decisive difference between these products consists in the functionality distribution, within a molecular weight range higher than 7000.     

藻类水热液化产物生物油分离纯化及组分分析

杜氏盐藻通过水热液化制备得到的生物油,先采用溶剂分割分别得到正己烷相、二氯甲烷相和乙醚相,再对二氯甲烷相进行柱层析分离纯化。正己烷相和乙醚相直接通过GC-MS和FT-IR进行分析,二氯甲烷相先经柱层析分离后结合GC-MS、二次质谱和FT-IR等确认不同馏分的产物组成。研究结果表明:二氯甲烷相经柱层析分离可得到16个馏分,分别是石油醚馏分(A₁),主要是烯烃类;石油醚:乙酸乙酯馏分(A₂),主要是酸类化合物;石油醚:丙酮馏分(A₃),主要是酰胺类;石油醚:甲醇馏分(A₄),主要是烷烃类;甲醇馏分(A₅),主要是十八碳烯酰胺。经柱层析分离纯化后,生物油的回收率高达91.38%;获取较全的生物油组分信息,为藻类液化机理的分析和生物油的改质提供了依据。     

Coupled column chromatography for copolymer characterisation

Coupled column chromatography has been performed with two chromatographic systems in which fractions from a size exclusion column are injected into a second column where retention is determined by non-exclusion mechanisms. This has been accomplished by directing the eluent from the first column system through the injection valve of the second column system. The first column system contained crosslinked polystyrene gel with the eluent tetrahydrofuran as good solvent. The second column system contained crosslinked polystyrene gel (or crosslinked polyacrylamide gel) with a mobile phase consisting of a solvent/non-solvent mixture which is a poor solvent promoting non-exclusion mechanisms for statistical copolymers of styrene and n-butyl methacrylate. Separations dependent on copolymer composition as a function of the non-solvent component in the mobile phase and the polarity of the stationary phase in the second column system have been accomplished by isocratic elutions.     

低压硅胶柱层析分离制备高纯辣椒素

从辣椒素总碱（Capsaicinoids）中提取高纯度的辣椒索及二氢辣椒素单体,建立了分离工艺条件。以TLC实验筛选适当的洗脱流动相,低压硅胶柱层析分离纯化,产物用两相法重结晶,TLC及HPLC监测分离效果及产品纯度。结果表明优选的流动相为混合溶剂石油醚：丙酮=8：2（体积比）,纯化分离得到辣椒素和二氢辣椒索,纯度分别从64％和29％提高到97％和96％。     

适应高滴度抗体制备的抗体捕集纯化技术研究进展

抗体捕集纯化是单克隆抗体和Fc融合蛋白药物制备过程的关键步骤。但相较于大规模动物细胞培养技术的迅猛发展，抗体的捕集纯化已经成为了制约单抗药物生产的主要“瓶颈”。本文回顾了蛋白A亲和层析和阳离子交换层析等当前抗体药物工业生产过程主要捕集纯化技术的发展现状和应用情况，介绍了近年来亲和肽层析和混合模式吸附层析等新型层析分离技术的发展以及膨胀床吸附和多柱串联吸附等过程集成方法在提高抗体捕集效率方面所展现的良好前景。在此基础上，指出了影响抗体捕集纯化的主要因素以及各技术发展中存在的问题，展望抗体捕集纯化技术的发展方向。     

Determination of functional type distribution of oligocaprolactonediols by liquid chromatography under critical conditions

Summary Functional type separation of oligocaprolactonediols by liquid chromatography of macromolecules under critical conditions was investigated. The analysis was performed with the aid of a novel variant of HPLC in the critical region on the boundary of the exclusion and adsorption separation modes. This method is characterized by practically complete disappereance of separation by molecular masses thus giving only information on the type of functionality.The critical conditions have been found on two systems. The first consists of a Lichrospher Si 100 column and a hexane-acetone mixture as eluent, while the second consists of a Zorbax NH2 column and a diethylether-acetone mixture.     

HPLC of plasmalogen-containing phosphatidylcholine under reverse-phase or argentation conditions

Two approaches to the high pressure liquid chromatography (HPLC) isolation of intact plasmalogens were investigated. The first used reversed-phase HPLC and sought to take advantage of subtle differences in the hydrophobicity of the alk-1-enyl chain from the acyl counterpart. On a C-18 column, bovine heart phosphatidylcholine (PC), which was 47% plasmalogen, was separated into a number of fractions that differed in their molecular species composition. One combination of fractions amounted to a 26% yield of PC enriched to 82% plasmalogen. The second approach sought to take advantage of the uniquely electron-rich functionality of the plasmalogens, the alk-1-enyl ether double bond, and its potential to coordinate with heavy metal ions. Specificially, bovine heart PC was applied to a cation-exchange type HPLC column in the silver ion mode. Although complete exchange of all the active sites of the column with silver ion led to complete retention of PC, partial activation with silver ion resulted in the separation of the PC into fractions, according to the degree of unsaturation. Plasmalogen-rich fractions eluted last and remained intact during the process. One combination of these fractions amounted to a 49% yield of PC enriched to 72% plasmalogen. Use of a cation-exchange system in the mercuric ion mode led to on-column hydrolysis of the plasmalogen; with palladium ion, the metallic species was stripped from the column by the eluting lipid.     

Separation of squalene from palm fatty acid distillate using adsorption chromatography

A method to separate squalene from palm fatty acid distillate (PFAD) using neutralization‐hydrolysis‐neutralization before employing adsorption column chromatography was developed. Extraneous matters, especially free fatty acids (83.8%) and acylglycerols (12.7%), were first neutralized and removed before being subjected to hydrolysis by using commercially available, immobilized Candida antartica lipase, at 65 °C for 8 h. Neutralization followed by hydrolysis and repeated neutralization successfully concentrated squalene from an initial amount of 3.76% to 27.5%. Oil extracted from neutralized‐hydrolyzed‐neutralized PFAD was then passed through a Diaion HP‐20 column using reverse‐phase adsorption chromatography. Squalene was desorbed by hexane, with a recovery of 93%.     

重烷基苯磺酸的品质改善研究

采用硅胶吸附法和碳酸钡中和法,脱除重烷基苯磺酸（HABSA）中的硫酸。以石油醚和乙醇为洗脱剂,采用硅胶层析柱法洗脱分离HABSA与重烷基苯（HAB）。实验结果表明：硅胶吸附法虽然能脱除一定量的硫酸,但其HABSA回收效果差;与之相反,碳酸钡中和法可脱除70%以上的硫酸,且保持高的HABSA回收率。实验结果也表明,硅胶柱层析法是一种快速简便且高效的分析或工业分离HAB与HABSA的方法。碳酸钡中和法和硅胶柱层析法的联合运用可有效脱除HABSA中的杂质,达到品质改善的目的。     

Quantitative Papier-Chromatographie der Fettsäuren

Systematic Quantitative Analysis of Natural Waxes with the Help of Ion-Exchange, Column and Thin-Layer Chromatography A method was developed for the quantitative separation of natural waxes into substance classes. The hydrocarbons were separated by the adsorption chromatography on silica gel/CS₂ column and the separation of acids from alcohols was achieved on a two-phase ion-exchange column after saponification. The isolation of the individual homologues (C₁₆? C₃₂) from the alcohol and acid mixtures was carried out with the help of reversed phase column chromatography by stepwise increase in temperature and acetic acid concentration.     

Isolation of a Target Component from a Multi‐component Mixture by Coupling Batch and Continuous Countercurrent Chromatography

A typical situation in preparative chromatography is that a certain target component elutes somewhere in the middle of a train of other components present in the feed. In order to isolate efficiently this target component the operating conditions of conventional batch chromatography have to be optimized carefully. Nevertheless, the achievable production rates and recovery yields are frequently low. Recently a possible alternative was suggested [1] based on using batch chromatography just as a first step providing fractions where the target component has the highest or lowest retention time. Subsequently, these fractions could be processed using continuous countercurrent chromatography in order to isolate the target component. Such a coupled process is analyzed theoretically for the case of linear adsorption isotherms. Based on results of simulations using a) the ideal model of chromatography and b) the equilibrium dispersive model it is demonstrated that the coupled process arrangement could possess advantages, compared to the application of batch chromatography alone.     

Theory of chromatography of linear and cyclic polymers with functional groups

We discuss the chromatographic behavior of linear polymers and rings having specific (functional) adsorption-active groups. Functionalized eight-shaped, daisy-like and theta-shaped macromolecules are considered as well. By using a model of an ideal chain with point-type defects in a slit-like pore we derive equations for the distribution coefficient covering all modes of chromatography of functionalized polymers of any molar mass in both narrow and wide pores. Additionally simple approximate formulae are obtained for a number of important modes of chromatography; chromatograms are simulated for model mixtures of polydisperse non-functional and functional polymers. By using the theory we analyze separation of polymers by molar mass, functionality and topology. Although there is a principal possibility to use adsorption chromatography or size-exclusion chromatography (SEC), we conclude that the liquid chromatography at the critical condition (LCCC) is especially efficient for separation of polydisperse polymers by both functionality and topology. The theory predicts that functionalized linear and cyclic polymers can be separated from each others by LCCC even better than non-functionalized ones. The LCCC behavior of some other types of polymers such as comb-like and semicyclic ones is discussed as well.     

Separation of Amino-Acids by Reversed Phase Chromatography: Development of a Model Mixture Dedicated to the Study of Solvent Gradient Chromatography Processes

Solvent gradient is currently used in analytical chromatography and presents an increasing interest in preparative chromatography. The applications relate to complex separations of mixtures where the target species has an intermediate affinity for the solid support compared to that of the other species, which is often the case during the purification of biomolecules extracted from natural raw materials or resulting from biotechnologies. In this case, separation leads to three fractions, impurities weakly retained, an intermediate fraction, and impurities strongly retained. An inexpensive and non-toxic mixture, containing five amino-acids, was identified as a good model mixture for such studies. These amino-acids were selected by taking into account their non-polar and hydrophobic nature. They were selected because of their aptitude for being easily detected in UV-visible spectroscopy without derivation. Separations were carried out by reversed phase chromatography. An experimental study first enabled determining the adsorption isotherm parameters of each amino-acid on an analytical column then on a preparative column for various organic solvent fractions. An empirical law was chosen to express the retention factor k as a function of the mobile phase composition (k=f (x_M)). An approach based on the column modeling by mixed cells in series enabled to simulate the separations obtained on the two types of columns. The use of the laws giving retention factor k as a function of the organic solvent fraction x_M then made it possible to carry out simulations for various solvent gradients. A simple methodology was developed to calculate the optimal solvent gradient. The gradient was optimized by minimizing the separation time and by respecting a constraint on the peaks resolution of the two species which are the most difficult to separate. A really good adequacy was observed between the experimental simulations and results for the weakly retained fraction as well as for the intermediate fraction which comprises components difficult to separate. Adequacy is not as good for strongly retained fraction but it is worth using these two species in the model mixture. This model mixture proves to be interesting to study solvent gradient preparative chromatography processes. The retention of these compounds is sensitive to the organic solvent fraction. It is easy to get these products which are inexpensive, non-toxic, and detectable without any derivation.     

柱径对制备色谱性能影响的分析

制备色谱是得到高纯度药物中间体的极好手段。柱径的选择是制备色谱分离中的关键问题。在制备色谱的应用中,柱效和压力是两个重要的参数。因此,分析柱径对压力和柱效的影响具有重要意义。本文在基本测试的基础上,研究了柱径对柱压力和柱效的影响。结果表明,随柱径增大,柱的孔隙率减小;柱压力并不随柱径的增大,呈平方倍数的增加;柱效随柱径的增大而增大。     

A comparison of the effects of various purification treatments on the oxidative stability of squid visceral oil

Factors influencing the oxidative stability of refined squid visceral oil after various purification treatments were evaluated. Squid visceral oil was treated by refining, steam deodorizing, molecular distillation, Sep-Pak cartridge extraction, alumina adsorption, EDTA solution washing, and column chromatography, and the treated oils were compared for oxidative stability. Sep-Pak cartridge extraction and alumina adsorption were designed to eliminate phenolic compounds, α-tocopherol, phospholipids, and metal ions from the oil. The oxidative stability was measured using PV and increase in weight. The refined oil and the deodorized oil were the most stable, and the highly purified oil was the least stable. However, alumina-adsorbed oil that had been washed with EDTA was more resistant to oxidation than highly purified oil. When refined oil was passed through an activated carbon-Celite chromatography column, it could be separated into hexane, ether, and ethanol fractions. The ethanol eluate contained more α-tocopherol and phospholipids than the ether eluate. The addition of the ethanol eluate extracted from squid visceral oil to the highly purified oil resulted in excellent stability.     

Matrix‐Assisted Laser Desorption/Ionization Mass Spectrometry of Synthetic Polymers, 5. Analysis of Poly(propylene oxide)s by Coupled Liquid Chromatography at the Critical Point of Adsorption and MALDI‐TOF Mass Spectrometry

Poly(propylene oxide)s can be analyzed efficiently with respect to functionality and molar mass by coupled liquid chromatography and MALDI‐TOF mass spectrometry. Both techniques are coupled via a robotic interface, where the matrix is coaxially added to the eluate and spotted dropwise onto the MALDI target. It is shown that size exclusion chromatography and liquid chromatography at critical conditions of adsorption coupled to MALDI‐TOF yield useful structural information on oligomer masses and chemical composition. In particular, the analysis of technical poly(propylene oxide) triols by LC‐CC/MALDI‐TOF reveals the presence of diols and monools in addition to the triols. The oligomer distributions of all functionality fractions are determined.     

辣椒红素与辣椒碱的分离

将干辣椒粉碎,以甲醇作溶剂回流提取2次,每次2h,溶剂用量依次为原料质量的5倍、3倍,合并提取液,过滤,减压蒸馏至浸膏状。甲醇提取物以甲醇-水〔V(甲醇)∶V(水)=1∶1〕为洗脱剂进行硅胶柱层析,用薄层层析-碘化铋钾喷雾联合检测,收集辣椒碱部分,无水乙醇结晶得无色细针晶,熔点63～65℃。再用乙醇作溶剂洗脱辣椒红素,脱除溶剂得到红色油状辣椒红素产品,色价为150。     

The unusual properties of high mass materials from coal-derived liquids

This short paper highlights the unusual properties of the high-mass material of coal liquids isolated by their insolubility in pyridine and solubility in NMP. The separation has been achieved by a column chromatography method. One gram quantity have been processed and near quantitative recovery of the sample as fractions has been achieved. This fractionation permitted recourse to a broad range of analytical methods, including some (e.g. ¹³C NMR), which require large sample sizes. Multiple macro analyses have been undertaken, using elemental analysis, TGA proximate analysis, NMR and FT-ir in addition to the micro-analytical methods used previously—pyrolysis-gc-ms, SEC, UV-fluorescence, probe-ms and MALDI-ms. The fractions show increasing concentrations of large molecular mass material with increasing polarity of successive eluents used in the fractionation. Evidence from solid-state ¹³C NMR and UV-fluorescence spectroscopy show progressive structural changes with increasing apparent molecular mass.