Pressure-programming of a binary,on-line mixed mobile phase for capillary column SFC using a packed column SFC pumping system

Summary Mixed mobile phase delivery for CSFC is served by a system previously designed for packed column SFC, where a back-pressure regulator controls pressure programming. Piston pumps deliver separate flows of carbon dioxide and modifier (2-propanol). The on-line mixed phase with the injected sample is split to the capillary column. Compared to a commercially available CSFC instrument the system shows no significant differences in resolution. Applications show the advantages of using modifiers in CSFC, such as separations of polar, ionic and high molecular weight compounds, influences on selectivity and shorter retention times.     

Sampling techniques for SFC using peak focusing by pressure and/or temperature programming

In analogy to the focusing effective in capillary GC, performed with temperature programming but also with sectional cooling of the column inlet as in multidimensional capillary GC, peak focusing can easily be attained in SFC by adjustment of the mobile phase pressure as well as the column temperature. This may be of practical use in connection with sampling techniques giving poor, i.e. broad and unsymmetrical, peak shapes. Such disturbances may occur, for example, in time controlled valve sampling over longer switching times. Generally, all other negative influences on peak shape can be suppressed or compensated by trapping within the column inlet. Special trapping devices and “retention gaps” may also be coupled to the column inlet in order to create narrow starting plug widths. Positive pressure (density) and negative temperature programs give rise to peak compression besides the increase of peak capacity of the separation.     

Temperature programming elution in capillary supercritical fluid chromatography

Summary Effects of column temperature on the retention behaviour of aromatic hydrocarbons and dialkyl phthalates were investigated in capillary supercritical fluid chromatography (SFC) with carbon dioxide as the mobile phase. Negative temperature programming could partly replace pressure programming. Positive temperature programming was applicable to solutes with proper volatility, in which gas chromatography-like retention mechanism (partition process) was involved.     

A model for quantitatively describing linear velocity programming in capillary SFC

Summary Linear velocity in capillary SFC is commonly controlled by restricting capillaries. In this paper, a model is described that quantitatively predicts the linear velocity of a supercritical fluid in SFC using tapered or ceramic frittype restrictors. In this model, the flow from the restricting capillary is assumed to be an isentropic expansion. The variation of the linear velocity as a function of pressure, temperature and cross-sectional area of the restricting aperture was predicted by this model. This predictive capability is important to the use of gradient programming in capillary SFC. Finally, the ideal variable restrictor for gradient programming was found to be one that could reversibly increase or decrease the linear velocity independent of the pressure, temperature, and/or density conditions used to create the gradient.     

Pressure drop effects on selectivity and resolution in packed column supercritical fluid chromatography

The influence of pressure drop on retention, selectivity, plate height and resolution was investigated systematically in packed supercritical fluid chromatography (SFC) using pure carbon dioxide as the mobile phase. Numerical methods developed previously which enabled the prediction of pressure gradients, diffusivities, capacity factors, plate heights and resolutions along the length of the column were used for the model calculations. The effects of inlet pressure and supercritical fluid flow rate on selectivity and resolution are studied. In packed column SFC with pure carbon dioxide as the mobile phase, the pressure drop can have a significant effect on resolution. The flow rate is shown to have a larger effect than generally realized. The calculated data are shown to be in good agreement with the experimental results. Finally, the variation of the chromatographic parameters along a 5.5 meter long model SFC column is illustrated. The possibilities and limitations of using long packed columns in SFC are discussed. It is demonstrated that long columns with large plate numbers do not necessarily yield better separations.     

超临界流体色谱流程设计及其应用

本文设计了多功能超临界流体色谱流程,流程中包括毛细管/微填充柱SFC,GC,计算机控制温度、压力、密度及信号采集、处理,配置有超临界流体萃取池,解决了超临界流体色谱分流口易堵问题。利用该流程,将石腊、DC-200气相色谱固定相、黄油、蜂蜡、救心油、红花油等样品进行超临界流体色谱分离。     

The application of capillary SFC,packed column SFC,and capillary SFC-MS in the analysis of controlled drugs

Multi–component mixtures of controlled drugs, drug impurities, and adulterants have been analyzed by capillary SFC-FID, packed column SFC-UV, and capillary SFC-MS. Isocratic packed column SFC has been performed with binary and ternary mobile phases using a single syringe pump. The combination of capillary SFC and double focusing MS is described with reference to MS source pressures and the spectra obtained. The use of negative temperature programming in SFC is described.     

A study of polyethoxylated alkylphenols by packed column supercritical fluid chromatography

Alkylphenol polyethoxylates (APEs) are a widely used group of nonionic surfactants in commercial production. Characterization of the composition of APE mixtures can be exploited for the determination of their most effective uses. In this study sample mixtures contain nonylphenol polyethoxylates and octylphenol polyethoxylates. The separation of individual alkylphenols by ethoxylate units is performed by supercritical fluid chromatography (SFC)-UV as well as normal-phase high-performance liquid chromatographic (HPLC)-UV employing packed columns. The stationary phase and column length are varied in the SFC setup to produce the most favorable separation conditions. Additionally, combinations of packed columns of different stationary phases are tested. The combination of a diol and a cyano column is found to produce optimal results. An advantage of using packed columns instead of capillary columns is the ability to inject large amounts of sample and thus collect eluted fractions. In this regard, fractions from SFC runs are collected and analyzed by flow injection analysis-electrospray ionization-mass spectroscopy in order to positively identify the composition of the fractions. In comparing the separation of APE mixtures by SFC and HPLC, it is found that SFC provides shorter retention times with similar resolution. In addition, less solvent waste is produced using SFC.     

Rapid analysis using capillary supercritical fluid chromatography

Capillary supercritical fluid chromatography (SFC) is proving to be a viable and useful separation method for thermally labile and nonvolatile materials. As with other capillary chromatographic techniques, very fast separations can be accomplished by sacrificing total efficiency and optimizing the conditions for rapid analysis. This is achieved using short, small-bore capillary columns, increased mobile phase linear velocities and very fast pressure programming rates. These principles are demonstrated for the rapid separation of selected component systems.     

Supercritical fluid chromatography/mass spectrometry using a simple capillary-direct interface

Summary A simple method for interfacing capillary supercritical fluid chromatography with a commercial quadrupole mass spectrometer (SFC/MS) has been developed that yields good chromatographic peak shapes and good sensitivity. No modification of the mass spectrometer is required, and a single instrument can be used for both SFC/MS and GC/MS with conversion between modes requiring less than 20 min. SFC/MS separations and chemical ionization mass spectra of wax components, a triazine pesticide metabolite, abietic acid, and high molecular weight polycyclic aromatic hydrocarbons are reported.

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Kombination von Chromatographie mit überkritischen fluiden Phasen und Massenspektrometrie unter Verwendung eines einfachen direkten Capillar-Interface

     

超临界流体色谱对吴茱萸中吲哚类生物碱的快速分析

建立了超临界流体色谱快速分析吴茱萸中吲哚类生物碱的方法。以标准品混合物和复杂样品为对象比较4种色谱柱的分离效果,进行色谱柱的筛选;考察了进样体积、改性剂、添加剂、温度和背压对保留行为的影响。结果表明,进样体积对峰形影响显著;添加剂对保留时间和色谱峰形影响有限;改变改性剂能使保留时间显著改变;降低温度,升高背压,保留时间减小。经过优化,确定采用Waters ACQUITY UPC² BEH色谱柱,以甲醇为改性剂,在35 ℃柱温和2.07×10⁷ Pa背压条件下,15 min内完成复杂样品的分析。同时采用超高效液相色谱完成复杂样品的快速分析。结果表明,超临界流体色谱可用于天然产物的高效快速分析,同时该方法与超高效液相色谱在分离选择上的差异有助于天然产物分析方法的拓展。     

Fast supercritical fluid chromatography of polymers using packed capillary columns

Summary Fast separations of perfluorinated polyethers and polymethylsiloxanes that are composed of 50–80 oligomers were demonstrated in packed capillary column supercritical fluid chromatography (SFC) using a carbon dioxide mobile phase. Separations were accomplished within 10 min using a 13 cm×250 μm i.d. column packed with 2 μm porous octadecyl bonded silica (ODS) particles. Effects of particle diameter of the packing material and pressure programming on separation were investigated, and packed column SFC was compared with open tubular column SFC. Results show that as the particle diameter was decreased from 5 to 3 to 2 μm and the column length was reduced from 85 to 43 to 13 cm, the separation time could be reduced from 70 to 20 to 10 min while still maintaining similar separation (resolution). Short columns packed with small porous particles are very suitable for fast SFC separations of polymers.     

Rapid and high resolution capillary supercritical fluid chromatography (SFC) and SFC/MS of trichothecene mycotoxins

The potential application of capillary column supercritical fluid chromatography (SFC) and SFC/mass spectrometry (SFC/MS) for the separation and analysis of mycotoxins of the trichothecene group was examined. Trichothecenes present significant analytical problems for both gas and liquid chromatography with a major difficulty for the latter being the lack of sufficiently sensitive and selective detectors. Supercritical carbon dioxide mobile phases at temperatures up to 100 degrees C were used with deactivated fused silica columns coated with crosslinked stationary phases. Separations were obtained under pressure ramped conditions using long (15 m) 50-micron i.d. columns for several trichothecenes (diacetoxyscirpenol, deoxynivalenol, and T-2 toxin) and related higher molecular weight macrocyclic (roridin and verrucarin) trichothecenes. In addition, new rapid pressure programming techniques with short (less than 2m) 25- to 50-micron i.d. capillary columns were used to obtain fast separations in as little as 1 min. SFC/MS with ammonia chemical ionization provided high selectivity and sensitive detection (with approximately 1-pg detection limits) for trichothecene mixtures. The extension to complex sample matrices is discussed and the application of selective MS/MS detection is demonstrated.     

Retention behaviour of carboxylic acid methyl esters in supercritical fluid chromatography

The study on retention behavior in supercritical fluid chromatography (SFC) is necessary to understand the mechanism of the various interactions in SFC. The retention of SFC in carboxylic acid methyl ester/polymethylsiloxane/CO2 system was studied systematically and the retention behavior of this kind of compounds under various typical operation conditions was described using the method of an alternative unified theory of chromatographic retention. The results illustrated that expression: Ink.= a + b/T + cp + dp/T + ep2/T can be used to describe quantitatively the retention behavior of carboxylic acid methyl ester/polymethylsiloxane/CO2 system in the ranges of reduced density from 0.549 to 1.411. It was also found that the entropy of solute in stationary phase is dependent on the density of supercritical fluid (SF) under typical operating conditions of SFC.     

Chromatographic resolution of closely related species: Drug metabolites and analogs

In this study, we investigate the separation of a variety of mixtures of drugs, metabolites, and related analogs including representatives of the carbamazepine, methylated xanthine, steroid hormone, nicotine, and morphine families using several automated chromatographic method development screening systems including ultra high performance liquid chromatography, core–shell HPLC, achiral supercritical fluid chromatography (SFC), and chiral SFC. Of the 138 column and mobile phase combinations examined for each mixture, a few chromatographic conditions afford the best overall performance, with a single achiral SFC method (4.6 × 250 mm, 3.0 μm GreenSep Ethyl Pyridine, 25 mM isobutylamine in methanol/CO₂) affording good separation for all samples. Four of these mixtures were also resolved by achiral SFC on the Luna HILIC and chiral SFC Chiralpak IB columns using methanol or ethanol with 25 mM isobutylamine as polar modifiers. Modifications of standard chromatography screening conditions afforded fast separation methods (from 1 to 5 min) for baseline resolution of all components of each of these challenging sets of closely related compounds.     

Determination of paraffins in food simulants and packaging materials by liquid chromatography with evaporative mass detection and identification of paraffin type by liquid chromatography/gas chromatography and fourier transform infrared spectroscopy

A liquid chromatographic method with evaporative mass detection (EMD) is described for the determination of paraffins in food contact materials that do not contain polyolefin oligomers, or paraffins migrating from these materials into fatty food simulants or certain simple foods. A normal-phase column operating at maximum column efficiency separates nonparaffinic and paraffinic materials without resolving the latter into individual components, and EMD is used to quantitate the paraffins. An on-line qualitative method that uses liquid chromatography/gas chromatography with flame ionization detection discriminates between paraffin waxes and oils in food contact materials, food simulants, and certain simple foods; a Fourier transform infrared spectrophotometric qualitative method also discriminates between waxes and oils, but is usually restricted to food contact materials that do not contain polyolefins and to migration experiments with organic solvents as fatty food simulants (with some other fatty food simulants, paraffin type must then be identified in the food contact material).     

Correlation of supercritical-fluid extraction recoveries with supercritical-fluid chromatographic retention data: A fundamental study

The possibility of using supercritical-fluid chromatographic retention data for examining the effects of operational parameters, such as pressure and flow rate, on the extraction characteristics in supercritical-fluid extraction (SFE) was investigated. A model was derived for calculating the extraction efficiency in SFE from retention data and peak shapes measured in supercritical-fluid chromatography (SFC). By performing the SFC experiments at the same pressure and temperature as the SFE extractions using the SFE extraction cell as the SFC column, an accurate prediction of extraction efficiencies could be made. Finally, the effects of matrix composition and analyte concentration on extraction efficiency were studied.     

Supercritical fluid chromatography: Retention, resolution, and gradient methods

Summary For applying gradient methods in supercritical fluid chromatography (SFC), the dependence of retention and resolution on the physical parameters, temperature, pressure/density, linear velocity, and mobile phase composition, is of particular relevance. This stems from the finding that the dependence of retention and resolution on physical parameters is more varied for SFC than for either gas or liquid chromatography. Thus, both retention and resolution tend to form maxima with temperature at constant pressure. In this communication, the dependence of capacity ratio k and resolution R on temperature and pressure is shown as three-dimensional plots. From these plots the general requirements for single and multiple gradients are discussed. In addition, equations are given which relate the contributions of vapor pressure and solvation to the measured capacity ratio k and selectivity .

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Chromatographie mit überkritischen fluiden Phasen: Retention, Auflösung und Gradientenmethoden

     

The contribution of the pressure drop to analyte retention in coupled column supercritical fluid chromatography of lipids

Summary This paper reports the qualitative and quantitative effects of the column pressure drop on the retention of lipid components in a serially coupled capillary column SFC system. The contribution of the pressure drop consists of two components, the density effect and the flow effect. The magnitude of the flow effect,i. e. the change in retention which results from changes in the flow-rate when column pressures are changed, is determined by the difference in single column analyte k values. The effect will be positive compared with the uncorrected retention values when the column with largest k value is closest to the injector. With the columns in reversed order, the effect will be negative. The contribution from the density effect always resulted in larger coupled column k values and was in most instances of more significance than the flow effect component. Values calculated with and without pressure drop correction have been compared and it has been shown that for most of the eighteen model lipid compounds investigated, the deviations from the experimental retention factors were smaller when pressure drop corrections were made.     

Supercritical fluid chromatography in drug analysis: a literature survey

The applications of supercritical fluid chromatography to the analysis of drugs have been carefully revised from the literature compiled in the Analytical Abstracts until March 1994. Easy-to-read tables provide useful information about the state-of-the-art and possibilities offered by SFC in pharmaceutical analysis. The tables comprise extensive data about samples analyzed, pharmaceutical principles determined, solvents used and sample quantity injected, supercritical fluids and modifiers employed, injection system, instrumentation, experimental conditions for chromatographic separations (density, pressure, flow, temperature), characteristics of columns employed (type, support, length, diameter, particle film thickness, stationary phase), detectors, type of restrictors, and also some analytical features of the methods developed (such as retention time, resolution, sensitivity, limit of detection and relative standard deviation).