Reservoirs, HPLC

FIGURE 4.11 Experimental setup for the dynamic ultrasound-assisted extraction of phenolic compounds from olive leaves and alperajo (C, extraction coil EC, extraction chamber ER, extract reservoir HPLC, high-performance liquid chromatography LC, leaching carrier PC, personal computer PP, peristaltic pump SV, selection valve UP, ultrasonic probe WB, thermostatic water-bath). 

Injecting the Sample The mechanism by which samples are introduced in capillary electrophoresis is quite different from that used in GC or HPLC. Two types of injection are commonly used hydrodynamic injection and electrokinetic injection. In both cases the capillary tube is filled with buffer solution. One end of the capillary tube is placed in the destination reservoir, and the other is placed in the sample vial. 

Figure4.62 Experimental set-up for liquid/liquid experiments (a) reservoir for the substrate in n-heptane (b) water reservoir (c, d) high-pressure liquid pumps (e) HPLC injection valve with sample loop for catalyst injection (f) micro mixer ...

Figure 5.28 Schematic of the experimental set-up. Water/ethylene glycol/SDS reservoir (a) high-pressure liquid pumps (b) catalyst/ substrate HPLC injection valve with 200 pi sample loop (c) hydrogen supply, equipped with mass flow controller (d) micro mixer (e) heating jacket (f) tubular glass or quartz reactor (g) back-pressure regulator (h) [64],...

A sensitive and selective method, which is specific for oxamyl and may be applicable to other oxime carbamates and their metabolites in heavily polluted waters, involves the use of HPLC/MS/MS. A 75-mL reservoir is attached on top of a Bond-Elut SAX (l-g/6-mL) (Varian), which is connected to an Oasis HLB (l-g/20-mL)... 

One of the major practical problems to the installation of HPLC as a permanent process monitor is the need to replace solvent. A large solvent reservoir may present problems both in terms of size and safety. One solution is the use of packed capillary columns, which consume much less solvent than conventional columns, as the comparison (at constant linear velocity) in Table 1 shows. 

A modern solvent delivery system consists of one or more pumps, solvent reservoirs, and a degassing system. HPLC pumps can be categorized in several ways by flow range, driving mechanism, or blending method. A typical analytical pump has a flow range of 0.001-10 mL/min, which handles comfortably the flow rates required for most analytical work (e.g., 0.5-3 mL/min). Preparative pumps can have a flow range from 30 mL/min up to L/m. 

As the name implies, capillary electrophoresis is electrophoresis that is made to occur inside a piece (50 to 100 cm) of small-diameter capillary tubing, similar to the tubing used for capillary GC columns. The tubing contains the electrolyte medium, and the ends of the tube are dipped into solvent reservoirs, as is the paper in paper electrophoresis. Electrodes in these reservoirs create the potential difference across the capillary tube. An electronic detector, such as those described for HPLC (Chapter 13), is on-line and allows detection and quantitative analysis of mixture components. 

The HPLC pump draws the mobile phase from the reservoir via vacuum action. In the process, air dissolved in the mobile phase may withdraw from the liquid and form bubbles in the flow stream unless such air is removed from the liquid in advance. Air in the flow stream is undesirable because it can cause a wide variety of problems, such as poor pump performance or poor detector response. Removing air from the mobile phase, called degassing, in advance of the chromatography is a routine matter, however, and can be done in one of several ways 1) helium sparging, 2) ultrasonic agitation, 3) drawing a vacuum over the surface of the liquid, or 4) a combination of numbers 2 and 3. 

Examine the HPLC instrument to which you are assigned. Find the inlet line to the pump and place the free end of this line in the reservoir containing the mobile phase with the 90/10 composition. Trace the path of the mobile phase from the reservoir, through the pump, injection valve, column, and detector, to the waste container so that you identify and recognize all components of the flow path. Turn on the pump and detector and begin pumping the mobile phase at a rate... 

Detail the path of the mobile phase through the HPLC system from the solvent reservoir to the waste receptacle, giving brief explanatory descriptions of instrument components along the way. 

A reciprocating piston pump is a pump that utilizes a piston in a cylinder to pull and push the liquid mobile phase from the mobile phase reservoir through the HPLC system. Two check valves (backflow preventers) are in-line to help force the liquid in only one direction. See Figure 13.4. 

The gradient elution method for HPLC is the method in which the mobile phase composition is changed in some preprogrammed way in the middle of the run. The device that accomplishes this is called the gradient programmer and is placed between the mobile phase reservoir and the pump. It is useful in experiments in which altering the mobile phase composition assists with the resolution of the mixture. 

High Performance Liquid Chromatography (HPLC) (Chapter 30) gives an elaborate discussion of theoretical aspects. Instrumentation encompasses the various important components e.g., solvent reservoir and degassing system pressure, flow and temperature pumps and sample injection system ... 

Figure 3. Set-up of a correlation HPLC system. The constant water flow is controlled by a PRBS pattern which directs the flow to either the sample or the eluent reservoir driving the appropriate plunger forward. A 6-way rotary valve is placed at the outlet of the eluent reservoir to allow single injection experiments.

There are typically six components in an HPLC system (1) solvent reservoirs (2) a pumping or solvent management system (3) an injector, which can be either manual or automated (4) a column (5) a detector (6) a data recorder, which can be an integrator or a computer system. 

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