Injection splitter

Recently, Alexander et al. [44] reported the use of an automated separation system developed for micro-LC and CEC using both isocratic and gradient elutions. The complete system is shown in Fig. 2.15. An enlarged view of the coupling of the column to the injection valve presents Fig. 2.16. The mobile phase was delivered by two micro-LC pumps at a flow rate of 30 pL/min to a post injection splitter that houses the column inlet. In the CEC mode, pressure was not applied (no restriction on... 

Behnke and Bayer published a similar approach for gradient elution PEC [51]. The schematic view is shown in Fig. 2.17. A gradient mixer and a HPLC pump were combined with a modular CE system. A post-injection splitter was used and sample introduced by a conventional HPLC six-port injector. A grounded stainless-steel T-piece was used to split both eluent and sample. The electrolyte reservoir on the inlet side of the separation capillary was connected to the splitter by a homemade interface. 

Capillary columns GC columns, e.g. WCOT, PLOT, having a bore of 0.1 -0.7 mm with a 0.1 -5 pm film of stationary phase coated onto the inner wall of a pure silica glass column 10-50 m long, high column efficiency, N ff, and excellent resolution are achieved. Capacity of the stationary phase is very low so a sample injection splitter is used to introduce approximately 0.01 pi of sample onto the column. 

Sample injection splitter used in capillary GC to permit small samples to be introduced onto the column, e.g. for a split ratio of 1 100 and a 1 pi injection 0.01 pi would enter the column. 

S ample loop volumes The volumes of the sample loops that store eluent from the first dimension and inject eluent into the second-dimension column system must be determined. The loop volume divided by the second-dimension elution time range determines the first-dimension flow rate in comprehensive 2DLC. If the dilution factor is small in the second column, a flow splitter can maintain a small loop volume even with a substantial flow rate from the first-dimension column. 

Cone splitter, as shown in Fig. 25, used in general injection applications for up to 8-way splitting and claimed (Hilbert, 1982) to achieve 10% accuracy in splitting. It should be noted that such figures depend more on the design of the pipe branches downstream of the splitter, rather than the splitter itself. 

Rotary splitter (Selves and Barnes, 1993), which can be used to provide up to 36-way splitting. Due to its intermittent operation, the pulsing flow in the branches downstream of the splitter would not be suitable for applications requiring a smooth and regular injection of material. 

These have now been superseded by capillary columns, which offer greatly improved separation efficiency. Fused silica capillary tubes are used which have internal diameters ranging from 0.1 mm (small bore) to 0.53 mm (large bore) with typical lengths in excess of 20 m. The wall-coated open tubular (WCOT) columns have the internal surface of the tube coated with the liquid (stationary) phase and no particulate supporting medium is required. An alternative form of column is the porous-layer open tubular (PLOT) column, which has an internal coating of an adsorbent such as alumina (aluminium oxide) and various coatings. Microlitre sample volumes are used with these capillary columns and the injection port usually incorporates a stream splitter. 

Operation of the column oven at 50°C or lower has been a problem in earlier chromatographs because of the difficulty of completely isolating the column oven from other heated components, such as the detector, injection port, and splitter, and still having a usable oven. The processor controller described overcomes this problem by mixing controlled amounts of room air into the column oven and can control very adequately at temperatures of about 30°C without cryogenic cooling. A further advantage of the processor controller is that the processor normally also can handle the temperature control of the other heated zones—inlet, detector, valves, and so on. 

A single Sample Simultaneously Injected into Two Dissimilar Columns. This is done by means of a simple inlet splitter which allows half the sample to enter one column and the other half of the sample to enter the second column. Two similar... 

An HPLC chromatograph was used together with a mass detector (when required, a stream splitter (app. 10 1) was inserted between the column and the detector). A column (250 X 4.6-mm ID) of NUCLEOSIL 5SA was flushed with 1% aqueous ammonium nitrate solution at a flow rate of 0.5 ml/min for 1 h, then with distilled water at 1 ml/min for 1 h. Silver nitrate (0.2 g) in water (1 ml) was injected onto the column in 50-/zl aliquots at 1-min intervals silver began to elute from the column after about 10 min. Twenty minutes after the last injection, the column was washed with methanol for 1 h, then with 1,2-dichloroethane-dichloromethane (1 1) for another hour. The three solvent reservoirs contained the following (A) 1,2-dichloroethane-dichloromethane (1 1) (B) acetone and (C) acetone-acetonitrile (9 1). For linoleic acid-rich seed oils, gradients of A were employed to 50% A-50% B over 15 min, then to 50% B-50% C over a further 25 min and held there for 5 min. For linolenic acid-rich seed oils, C was changed to acetone-acetonitrile (4 1), and the flow rate was increased to 1 ml/min gradients of A were utilized to 50% A-50% B over 10 min, then to 70% B-30% C over 20 more min, and finally to 100% C over another 30 min. 

Experimental (simplex and window diagram). The chromatographic system consisted of a Model 501 supercritical fluid chromatograph (Lee Scientific, Salt Lake City, Utah) with the flame ionization detector (FID) set at 375°C. The instrument was controlled with a Zenith AT computer. A pneumatically driven injector with a 200 nL or a 500 nL loop was used in conjunction with a splitter. Split ratios used were between 5 1 and 50 1, depending on sample concentration and the chosen linear velocity, while the timed injection duration ranged from 50 ms to 1 s. We found that the variation of both the split ratio and injection time allowed greater control over the... 

Analytical Instrumentation A Hewlett Packard model 5880-A series gas chromatograph equipped with flame ionization detection (FID) was fitted with an 18 m x 0.25 mm id glass capillary column coated with OV-101. Reaction mixtures and standards were injected through a stainless steel injector splitter with a split ratio of 100 1. The column oven was linearly programmed from 90 to 200°C at 2 or 5°/min. The injector and detector temperatures were... 

Preparative separations in the grams per injection level are different. Separations are run isocratic in 1- to 3-in columns with large pore, fully porous packings (35-60jUm). An analytical, two-pump system can just barely reach the 20-mL/min flow rates needed to run a 1-in column. Special preparative HPLC systems deliver flow rates of 50-500 mL/min to handle the larger bore columns. A stream splitter is used to send part of the flow through a refractive index detector with a flow cell designed for concentrated solutions. 

Sample introduction is a major hardware problem for SFC. The sample solvent composition and the injection pressure and temperature can all affect sample introduction. The high solute diffusion and lower viscosity which favor supercritical fluids over liquid mobile phases can cause problems in injection. Back-diffusion can occur, causing broad solvent peaks and poor solute peak shape. There can also be a complex phase behavior as well as a solubility phenomenon taking place due to the fact that one may have combinations of supercritical fluid (neat or mixed with sample solvent), a subcritical liquified gas, sample solvents, and solute present simultaneously in the injector and column head [2]. All of these can contribute individually to reproducibility problems in SFC. Both dynamic and timed split modes are used for sample introduction in capillary SFC. Dynamic split injectors have a microvalve and splitter assembly. The amount of injection is based on the size of a fused silica restrictor. In the timed split mode, the SFC column is directly connected to the injection valve. Highspeed pneumatics and electronics are used along with a standard injection valve and actuator. Rapid actuation of the valve from the load to the inject position and back occurs in milliseconds. In this mode, one can program the time of injection on a computer and thus control the amount of injection. In packed-column SFC, an injector similar to HPLC is used and whole loop is injected on the column. The valve is switched either manually or automatically through a remote injector port. The injection is done under pressure. 

Due to the high sensitivity it is favorably to couple a nanoHPLC to an ESI-source. As mass spectrometers are concentration dependent detectors, the sensitivity of an instrumental setup is mostly determined by the peptide concentration of the eluate but not by the peptide amount. Thus a nanocolumn with a flow rate of 300 nL/min provides an about thousand times higher sensitivity than a microbore column with a flow rate of 300 (xL/min. As an alternative to buying a nanoHPLC system it is also possible to use a relatively inexpensive flow splitter after the pump and before the injection valve and the column. Thereby the flow rate can be reduced to use a capillary column (flow rate 4 (xL/min) on an analytical HPLC system or a nanocolumn on a capillary HPLC system. Instead of a flow-splitter it is preferred to couple a nanoHPLC to an ESI-source. Thereby, the flow rate is split according to the column backpressure, i.e., mostly the column volumes if the same packing materials are used. However, these low-cost setups are less reliable than a nanoHPLC and the reproducibility is worse. 

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