Micro flow chromatography

The mass spectrometer is a very sensitive and selective instrument. However, the introduction of the eluent into the vacuum chamber and the resulting significant pressure drop reduces the sensitivity. The gas exhaust power of a normal vacuum pump is some 10 ml min-1 so high capacity or turbo vacuum pumps are usually needed. The gas-phase volume corresponding to 1 ml of liquid is 176 ml for -hexane, 384 ml for ethanol, 429 ml for acetonitrile, 554 ml for methanol, and 1245 ml for water under standard conditions (0°C, 1 atmosphere). The elimination of the mobile phase solvent is therefore important, otherwise the expanding eluent will destroy the vacuum in the detector. Several methods to accomplish this have been developed. The commercialized interfaces are thermo-spray, moving-belt, electrospray ionization, ion-spray, and atmospheric pressure ionization. The influence of the eluent is very complex, and the modification of eluent components and the selection of an interface are therefore important. Micro-liquid chromatography is suitable for this detector, due to its very small flow rate (usually only 10 p min - ). 

Chervet, J. R, Meijvogel, C. J., Ursem, M., and Salzmann, J. P, Recent advances in capillary liquid-chromatography—Delivery of highly reproducible micro flows, LC GC—Magazine of Separation Science 10(2), 140-148, 1992. 

Zhou, X., Eurushima, N., Terashima, C., Tanaka, H., and Kurano, M., New micro-flow pumping system for liquid chromatography. Journal of Chromatography A 913(1-2), 165-171, 2001. 

Chassaing, C., Stafford, H., Luckwell, J., Wright, A., and Edgington, A. (2005). A parallel micro turbulent flow chromatography-tandem mass spectrometry method for the analysis of a pharmaceutical compound in plasma. Chromatographia 62 17-24. 

The University of Washington group, Christian et al. (69), recently introduced a unique micro-flow-through cell for fluorescence determination of effluents from liquid chromatography columns. A laminar flow design reduces scatter and minimizes dead volume. The sample volume can be maintained at less than 100 nL with a wide range of flow rates. 

Table 21.1 classifies popular LC detectors according to several criteria for purposes of comparison. At the present time, LC detectors are generally less sensitive than GC detectors, which can detect picograms of material under good conditions. Most LC detectors provide only limited structural information. However, spectrophotometers fitted with micro flow-cells can be used to obtain a stop-flow ultraviolet-or visible-absorption spectrum of an LC peak trapped in the flow cell. On-line coupling of liquid chromatographs with mass or infrared spectrometers offers sophisticated, but indeed expensive, detection/identification methods. Such systems have been described in the literature, but are quite limited by the solvents that can be used in the chromatography step. 

An evaluation of the scientific literature reveals that over 500 papers on veterinary drug residue analysis were published in the 5-year period of 2005-2009. Liquid extraction (LE) and liquid-solid extraction (LSE) were found to be very popular sample treatment techniques that were used in 30% and 60% of the reported studies, respectively. Here, LE includes all liquid-based approaches such as liquid-liquid extraction (LLE), extrelut liquid-liquid extraction, liquid-liquid micro-extraction, and pressurized liquid extraction (PLE). LSE includes solid phase extraction (SPE) and all other sorbent-based extraction procedures, such as solid phase micro-extraction (SPME), stir bar sorptive extraction (SBSE), restricted-access materials (RAM), turbulent-flow chromatography (TEC), dispersive SPE (dSPE), and matrix solid phase dispersion... 

Ethylbenzene disproportionation catalyzed by add zeolites was studied by Karge et al. [887, 888] and recommended as a versatile test reaction for acid (monofunctional or bifunctional) catalysts such as add zeolites or related materials and is frequently used for this purpose. Also, this reaction was studied in situ by IR spectroscopy in combination with gas chromatography for determination of conversion and selectivity [888]. In these experiments, a flow-reactor quartz glass cell as described in Ref. [152] was used, which could be operated imder ultra-high vacuum during the pretreatment of the thin catalyst wafers of pressed zeolite powder at, e.g., 670-870 K and 10 Pa. After pretreatment, the cell was used as a differential fixed-bed micro-flow reactor (cf. also [ 152,158]). Results are illustrated by Fig. 52. 

Turnpenny P, Fraier D, Chassaing C, Duckworth J. Development of a micro-turbulent flow chromatography focus mode method for drug quantitation in discovery bioanalysis. J Chromatogr B Anal Technol Biomed Life Sci 2007 856 131-140. 

A technique for resolving and determining the activities of the components of jS-D-acetamidodeoxyhexosidase, based on micro-column chromatography on DEAE-cellulose, has been described. The column eflSuent is led directly into the flow-stream of an autoanalyser and mixed with a buffered solution of a fluorogenic substrate (4-methylumbelliferyl 2-acetamido-2-deoxy-j8-D-gluco-pyranoside) to produce a continuous trace of the elution profile of the enzymic... 

These factors make it necessary to reduce the amount of solvent vapor entering the flame to as low a level as possible and to make any droplets or particulates entering the flame as small and of as uniform a droplet size as possible. Desolvation chambers are designed to optimize these factors so as to maintain a near-constant efficiency of ionization and to flatten out fluctuations in droplet size from the nebulizer. Droplets of less than 10 pm in diameter are preferred. For flow rates of less than about 10 pl/min issuing from micro- or nanobore liquid chromatography columns, a desolvation chamber is unlikely to be needed. 

Ion chromatography (see Section 7.4). Conductivity cells can be coupled to ion chromatographic systems to provide a sensitive method for measuring ionic concentrations in the eluate. To achieve this end, special micro-conductivity cells have been developed of a flow-through pattern and placed in a thermostatted enclosure a typical cell may contain a volume of about 1.5 /iL and have a cell constant of approximately 15 cm-1. It is claimed15 that sensitivity is improved by use of a bipolar square-wave pulsed current which reduces polarisation and capacitance effects, and the changes in conductivity caused by the heating effect of the current (see Refs 16, 17). 

Principles and Characteristics As mentioned already (Section 3.5.2) solid-phase microextraction involves the use of a micro-fibre which is exposed to the analyte(s) for a prespecified time. GC-MS is an ideal detector after SPME extraction/injection for both qualitative and quantitative analysis. For SPME-GC analysis, the fibre is forced into the chromatography capillary injector, where the entire extraction is desorbed. A high linear flow-rate of the carrier gas along the fibre is essential to ensure complete desorption of the analytes. Because no solvent is injected, and the analytes are rapidly desorbed on to the column, minimum detection limits are improved and resolution is maintained. Online coupling of conventional fibre-based SPME coupled with GC is now becoming routine. Automated SPME takes the sample directly from bottle to gas chromatograph. Split/splitless, on-column and PTV injection are compatible with SPME. SPME can also be used very effectively for sample introduction to fast GC systems, provided that a dedicated injector is used for this purpose [69,70],... 

The current status of chromatography is shown in Table 10.25. Since reducing separation time is a major issue, there is a pronounced trend toward shorter columns filled with small particles. The current trends for lower flow (micro- and nano-LC) columns, and great strides to achieve (ultra-) fast chromatographic... 

After the activation period, the reactor temperature was decreased to 453 K, synthesis gas (H2 CO = 2 1) was introduced to the reactor, and the pressure was increased to 2.03 MPa (20.7 atm). The reactor temperature was increased to 493 K at a rate of 1 K/min, and the space velocity was maintained at 5 SL/h/gcat. The reaction products were continuously removed from the vapor space of the reactor and passed through two traps, a warm trap maintained at 373 K and a cold trap held at 273 K. The uncondensed vapor stream was reduced to atmospheric pressure through a letdown valve. The gas flow was measured using a wet test meter and analyzed by an online GC. The accumulated reactor liquid products were removed every 24 h by passing through a 2 pm sintered metal filter located below the liquid level in the CSTR. The conversions of CO and H2 were obtained by gas chromatography (GC) analysis (micro-GC equipped with thermal conductivity detectors) of the reactor exit gas mixture. The reaction products were collected in three traps maintained at different temperatures a hot trap (200°C), a warm trap (100°C), and a cold trap (0°C). The products were separated into different fractions (rewax, wax, oil, and aqueous) for quantification. However, the oil and wax fractions were mixed prior to GC analysis. 

New concepts presented in this edition include monolithic columns, bonded stationary phases, micro-HPLC, two-dimensional comprehensive liquid chromatography, gradient elution mode, and capillary electromigration techniques. The book also discusses LC-MS interfaces, nonlinear chromatography, displacement chromatography of peptides and proteins, field-flow fractionation, retention models for ions, and polymer HPLC. 

Toluene disproportionation was carried out in a high-pressure continuous flow micro reactor. Granular catalyst (32-64 mesh, 2.5 cm ) was loaded into a stainless steel tube reactor. Toluene was fed at a rate of 10 cm h (liquid) in the flow of H2S(0.2vol.%)/H2 mixture gas (200 cm min b at 623K and 6MPa. The effluent was analyzed by gas chromatography (Shimadzu, GC-9A) by a flame ionization detector (FID). 

Micro- and nano-HPLC systems (Fig. 15.11) rely on small-diameter and capillary columns packed with high-efficiency packing materials and very slow flow rates to produce concentrated solutions and sharp chromatography peaks to feed electrospray interfaces for mass spectrometers. 

Analytical interfaces are integrated into the AuMpRes set-up for at-line analysis by sampling and subsequent chromatography (HPLC) [108], Moreover, this allows online analysis by infrared or Raman spectroscopy. Real-time monitoring of the chemical processes can be achieved via spectroscopic measurements, for which suitable optical flow-through cells have to be installed at selected positions of the micro reaction system. 

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