Instrumentation chromatography HPLC

Size-exclusion chromatography can be carried out using conventional HPLC instrumentation, replacing the HPLC column with an appropriate size-exclusion column. A UV/Vis detector is the most common means for obtaining the chromatogram. 

The basic instrument required for packed-column unified chromatography is shown schematically in Figure 7.9. This is essentially a two-pump HPLC instrument utilizing high-pressure mixing with just a few new components. At least one pump must... 

Many HPLC instruments are already furnished with temperature controls for the column. Unified chromatography requires a much wider temperature range than is currently practiced in HPLC. Until better defined by experience, a temperature range from about —60 to about 350°C seems reasonable as a specification. Since this is well in the range of a GC oven with subambient temperature capability, no new technology is required. 

Most widely used is the two-dimensional combination of SEC and HPLC for copolymer characterization. The typical HPLC instrument is very similar to an SEC apparatus. While the ideal SEC separation is exclusively determined by entropy changes, in HPLC or adsorption chromatography it is assumed that no... 

The mobile phase in HPLC is called the eluent and is a liquid or a mixture of liquids. Common eluents are water, aqueous solutions, acetonitrile, and methanol. Almost any other common solvent compatible with the column packing and the detector may be used. In some cases, the HPLC instrument will be capable of making a mixture of eluents or changing the mixture of eluents during chromatography. If this is done, care must be taken to make sure that the eluent mixture is compatible with the detector. 

For many years, the concept of the conductivity detector could not work, however. Ion chromatography experiments utilize solutions of high ion concentrations as the mobile phase. Thus, changes in conductivity due to eluting ions are not detectable above the already high conductivity of the mobile phase. This was true until the invention of so-called ion suppressors. Today, conductivity detectors are used extensively in HPLC ion chromatography instruments that also include suppressors. 

HPLC (high-performance liquid chromatography) is an instrumental chromatography technique in which the mobile phase is a liquid. Refer to Figure 13.1 and the accompanying discussion for details of the HPLC system. 

HPLC instrument equipped with a UV-vis detector column chromatography equipment. 

This chapter presents an overview of current trends in high-pressure liquid chromatography (HPLC) instrumentation focusing on recent advances and features relevant to pharmaceutical analysis. Operating principles of HPLC modules (pump, detectors, autosampler) are discussed with future trends. 

High-performance liquid chromatography (HPLC) is one of the premier analytical techniques widely used in analytical laboratories. Numerous analytical HPLC analyses have been developed for pharmaceutical, chemical, food, cosmetic, and environmental applications. The popularity of HPLC analysis can be attributed to its powerful combination of separation and quantitation capabilities. HPLC instrumentation has reached a state of maturity. The majority of vendors can provide very sophisticated and highly automated systems to meet users needs. To provide a high level of assurance that the data generated from the HPLC analysis are reliable, the performance of the HPLC system should be monitored at regular intervals. In this chapter some of the key performance attributes for a typical HPLC system (consisting of a quaternary pump, an autoinjector, a UV-Vis detector, and a temperature-controlled column compartment) are discussed [1-8]. 

The method of complete electrolysis is also important in elucidating the mechanism of an electrode reaction. Usually, the substance under study is completely electrolyzed at a controlled potential and the products are identified and determined by appropriate methods, such as gas chromatography (GC), high-performance liquid chromatography (HPLC), and capillary electrophoresis. In the GC method, the products are often identified and determined by the standard addition method. If the standard addition method is not applicable, however, other identification/determination techniques such as GC-MS should be used. The HPLC method is convenient when the product is thermally unstable or difficult to vaporize. HPLC instruments equipped with a high-sensitivity UV detector are the most popular, but a more sophisticated system like LC-MS may also be employed. In some cases, the products are separated from the solvent-supporting electrolyte system by such processes as vaporization, extraction and precipitation. If the products need to be collected separately, a preparative chromatographic method is use-... 

The differentiating feature of an open versus closed system is the ability to interact with and maximize the use of individual components. The closed system, on the other hand, represents a turnkey approach which enables full automation of the entire process. However, this instrument does not allow the user to interact with the individual system components dining processing. An example of a closed system is an HPLC instrument while flash chromatography represents an open system. 

---