Ion mobility detection after supercritical fluid chromatography

Using a unidirectional flow ion mobility detector, non-selective detection, tunable selective detection, and complete Fourier transformed ion mobility spectra were successfully obtained after supercritical fluid chromatography on compounds with higher molecular weights than have been previously investigated. In the most selective mode, single oligomers from polymeric material could be independently detected. Using the Fourier transform capabilities of this instrument, complete ion mobility spectra for each oligomer could be obtained in a single chromatographic separation. The collection of individual ion mobility spectra of the components of polymeric material has not been possible prior to the technique described in this paper. Only complex ion mobility spectra of polymeric mixtures are available in the literature. The spectra obtained in this study are all simple, uncomplicated spectra consisting of only one or two product ion peaks. K_o values reported in this work range from 0.633 to 1.61, which are some of the lowest values ever reported in ion mobility spectrometry. With the unidirectional flow design of the detector, the supercritical fluid mobile phase, carbon dioxide, was efficiently eliminated from the detector so that the ion mobility spectrometer could be operated in its normal manner. The fact that CO₂ did not interfere with normal ion mobility operation indicates that other supercritical fluids may also be compatible with this sensitive and versatile detection method.     

New detection strategies through supercritical fluid chromatography

This article reviews the current status and developmental trends in detection for supercritical fluid chromatography. The “GC-like” and “LC-like” detection techniques are treated separately, while the needs of structural identification techniques for SFC separations are emphasized.     

Peroxyoxalate chemiluminescence detection with packed column supercritical fluid chromatography

Summary A detection scheme based upon peroxyoxalate chemiluminescence, which utilizes two post-column pumps and two stages of depressurization is investigated. The chemiluminescent detection limit for perylene is 23 times lower than determined by fluorescence, and is in the attomole range. This detection technique is investigated for packed column supercritical fluid chromatography (SFC). Due to the interface design used and the chemical band narrowing effects of chemiluminescence, an apparent increase in efficiency is observed. The interface design affords a wide range of pressures to be used for a separation. During pressure programming the column effluent changes flow rate. Because of a back-pressure regulator, the reaction and detection take place at nearly constant pressure. Therefore pressure gradient work is possible without concern for post-column reagent solubility (which is a concern for high-performance liquid chromatography). The effects of the expanded CO₂ from the SFC on the chemiluminescence signal and background are studied. The post-column detection is optimized for pH, photomultiplier voltage, concentrations and flow rates of the peroxide and oxalate ester.     

Metal selective flame ionization detection after supercritical fluid chromatography

This work describes the construction and operation of a flame ionization detector for the selective detection of metal-containing compounds after capillary supercritical fluid chromatography. Using optimal conditions for achieving metal sensitive flame ionization responses which have been established for the detector after capillary gas chromatography, initial evaluation of the detector after SFC proved promising. Like the carbon sensitive FID, it appears that the metal sensitive FID is compatible with SFC when pure carbon dioxide is used as the mobile phase. Response characteristics were found to be similar to those observed when the detector is used in gas chromatography.     

Packed-capillary supercritical fluid chromatography of pesticides using phosphorus-selective detection

Summary Packed-capillary, supercritical fluid chromatography using carbon dioxide modified with methanol or 2-propanol as mobile phase, was coupled with a thermionic detector for phosphorus-selective detection. Optimum conditions for thermionic detection were established for a number of hydrogen flow rates by adjusting the air flow rate and bead position. Using the mentioned modifiers, a series of organophosphorus pesticides of varying polarity could be separated within 8 min. An optimum sensitivity of 55–128 fg P/s and linearity over four orders of magnitude were obtained. The detector showed good selectivity for phosphorus over carbon and nitrogen, viz., 9·10⁴ g C/g P and 75 g N/g P, respectively. This was demonstrated by determining pesticides such as phoxim, dimethoate and azinphos-methyl in onion and tomato extracts.     

Single pump on-line mixing system for capillary supercritical fluid chromatography

A simple inexpensive system comprising a piston pump, switching valves, and capillary tubing has been developed for preparing mixed mobile phases for capillary supercritical fluid chromatography. The system enables the on-line preparation of mixed mobile phases without contamination of the pump with modifier. Modifier concentrations, which can be changed stepwise, were determined both experimentally and theoretically, the latter on the basis of density–viscosity relationships. Examples of both isobaric and pressure programmed separations are demonstrated.     

Mobile phases for supercritical fluid chromatography

Summary Several liquids have been tested under supercritical conditions for their suitability in chromatographic separations. Experimental pressures were up to about 150 atm, temperatures to 300°C; stationary phases of differing polarity were used. Stability under all experimental conditions is shown by cyclohexane, di-i-propylether, benzene, tetrahydrofuran and ethylacetate, while halogenated alkanes, branched alkanes, ketones and acetonitrile decompose in some cases.     

An improved interface for universal acoustic flame detection in modified supercritical fluid chromatography

A novel method of interfacing the acoustic flame detector (AFD) with modified supercritical fluid chromatography (SFC) is presented. By applying resistive heating directly to the burner region between the restrictor outlet and the acoustic flame, infrequent severe noise, baseline drifting, and peak deformations that can occasionally be observed with the AFD are eliminated. For example, by increasing the interface temperature only a few hundred degrees Celsius, such sporadic noise in the detector can be reduced nearly ten-fold resulting in smooth stable operation of the AFD. Further, for various levels of methanol modified supercritical carbon dioxide mobile phase examined, the interface was observed to reduce detector noise in each to a common minimal range near 10-25 Hz when an appropriate temperature was achieved. The method is simply assembled, inexpensive to construct, and robust in its daily operation. Overall, the heated interface developed and presented facilitates reliable AFD operation in modified SFC, and supports further exploration and implementation of this sensor as an alternative universal detector in separations requiring an organic cosolvent in the mobile phase.     

Capillary supercritical fluid chromatography with flame photometric detection using a large bore column SFC pumping system

A commercially available instrument with an SFC pumping system suitable for wide bore columns (4.6 mm i.d.) has been modified for capillary supercritical fluid chromatography (CSFC) by incorporating a double-stage flow splitter. The first flow splitter was installed in front of the sample injection valve in order to avoid a high solute split ratio. The second splitter was mounted in the column oven so that the injected sample (0.2 μL) would be split to the capillary column. In order to perform pressure programmed elution, a pressure regulating system equipped with a gradient programmer has been used. Flame photometric detection was optimized for the analysis of organosulfur compounds by CSFC. In this study, detection limits were found to be 6–14 ng and the experimentally determined exponent (n value) varied from 1.721 to 1.984 depending on the compounds tested. Sulfur- and phosphorus-containing thermally labile pesticides can be chromatographed and selectively detected by using CSFC/FPD in either sulfur- or phosphorus mode, respectively.     

Development of a mass-directed preparative supercritical fluid chromatography purification system

In this paper, we report the development of a mass-directed supercritical fluid chromatography (SFC) purification system. We have addressed issues on software compatibility, the interface between the preparative SFC and the mass spectrometer, and fraction collection. Good peak shape and signal were achieved in the mass spectrometry (MS) trace, allowing accurate peak detection and reliable fraction collection. Simple modifications on a commercially available fraction collector enabled fractionation at atmospheric pressure with high recovery. The SFC/MS purification system has been used in support of high-throughput library purification and has been proven to be a valuable tool in complementing our reversed-phase high-performance liquid chromatograph (RP-HPLC/MS)-based technology platform.     

Large volume injection for preparative supercritical fluid chromatography

Summary A large volume injection system for preparative supercritical fluid chromatography is described. The method which is based on the solvent venting technique coupled with dilution of the sample solution consists of three steps. The first step is continuous dilution of the sample solution with liquid carbon dioxide at a controlled flow rate. The second step is solvent removal and solute trapping in a packed trap column. Combination of these two steps results in efficient solvent removal and the volume of sample which can be injected in a single injection becomes virtually unlimited. The third step is transfer and re-concentration of the solutes from the trap column on to the separation column with the pressures of both columns controlled independently; the final step is the separation. With this method, mass overloading behavior has been investigated and preparative separations performed.     

Solventless injecction for packed column supercritical fluid chromatography

The adverse effects of injection solvent strength on microbore packed column SFC band broadening are demonstrated and a solventless injection system that eliminates these effects is introduced. The injection system removes solvent in a GC-like manner using a retention gap and an on-column capillary GC syringe. The analyte is delivered to the analytical column in a solvent-free plug of supercritical fluid mobile phase.     

Capillary supercritical fluid chromatography with simultaneous flame ionization and mass spectrometric detection

A simple interface between a capillary supercritical fluid chromatograph and an Extranuclear Simulscan mass spectrometer is described. The SFC column is directly inserted into the ion source through the existing GC-interface. The system is equipped with a splitting device which allows simultaneous EI/MS and flame ionization detection when CO₂ is used as the supercritical phase. The effect of source temperature and pressure on CO₂ clustering was studied for optimization of source conditions. The performance of the system was evaluated with a series of model compounds and standard mixtures.     

Open-tubular supercritical fluid chromatography with simultaneous flame ionization and chemiluminescent nitrogen detection

The use of a simultaneous detection system for open-tubular supercritical fluid chromatogrphic separation of aniline isomers, alkyl(C₈ to C₁₈)dimethylamines, a dialkylmethylamine mixture, and an extract of a Japanese horseradish power “Wasabi” is demonstrated. A chemiluminescent nitrogen detector (CLND) and a flame ionization detector (FID) were configured at the end of the open tubular column via a zero- to dead-volume tee. The dual detection was achieved with a post-column split at a 1:2.5 ratio for the CLND and FID, respectively. The optimized CLND showed virtually no response to non-nitrogenous components such as benzene, toluene, and methanol which were used as injection solvents in these applications.     

Analysis of sulphonamides using supercritical fluid chromatography and supercritical fluid chromatography-mass spectrometry

Packed-column supercritical fluid chromatography has been used for the separation of mixtures of sulphonamides on silica and amino-bonded stationary phases utilizing carbon dioxide with methanol modifier as the mobile phase. The effect of modifier concentration, column pressure and modifier identity on retention was also studied. Packed-column supercritical fluid chromatography-mass spectrometry (SFC-MS) of these mixtures utilizing both moving-belt and modified thermospray interfaces was also studied. The identification of sulphamethazine in a spiked porcine kidney extract was performed by SFC-MS using the moving-belt interface.