High-performance liquid chromatography preparative column

The purity of crude peptide will be analayzed on a reverse-phase high-performance liquid chromatography (HPLC) column. The elution of the peptide is followed by absorbance recorded at 215 nm. The purification on a 10 to 20 mg scale is performed on a larger, preparative column monitored at the less sensitive wavelength of 238 nm. 

Trace enrichment is a sample precleaning procedure which is performed prior to a sample analysis. The purpose of any sample preparation procedure is twofold. First, such a procedure must selectively collect and concentrate the components of interest. Second, the method should eliminate any other components that would either interfere with the analysis or would contaminate an analytical gas chromatography (GC) or high-performance liquid chromatography (HPLC) column to shorten its useful analytical life. 

Purification schemes depend on the perceived nature of the impurities. The actual experimental conditions depend on the nature of the peptide. For each technique discussed, the user is encouraged to get in touch with the manufacturers of the media, high-performance liquid chromatography (HPLC) columns, equipment, etc. to acquire technical notes, which can provide very useful information on topics such as preparing media, solvent conditions, and HPLC strategies. 

SynChropak GPC supports were introduced in 1978 as the first commercial columns for high-performance liquid chromatography of proteins. SynChropak GPC columns were based on research developed by Fred Regnier and coworkers in 1976 (1,2). The first columns were only available in 10-yu,m particles with a 100-A pore diameter, but as silica technology advanced, the range of available pore diameters increased and 5-yu,m particle diameters became available. SynChropak GPC and CATSEC occasionally were prepared on larger particles on a custom basis, but generally these products have been intended for analytical applications. 

High performance liquid chromatography (HPLC) has been by far the most important method for separating chlorophylls. Open column chromatography and thin layer chromatography are still used for clean-up procedures to isolate and separate carotenoids and other lipids from chlorophylls and for preparative applications, but both are losing importance for analytical purposes due to their low resolution and have been replaced by more effective techniques like solid phase, supercritical fluid extraction and counter current chromatography. The whole analysis should be as brief as possible, since each additional step is a potential source of epimers and allomers. 

Colin, H., Hilaireau, P., and de Tournemire, J., Dynamic axial compression columns for preparative high performance liquid chromatography, LC-GC, 8, 302, 1990. 

Currently, all five toxic components from concavum are being subjected to column and high performance liquid chromatography. Products of these treatments will be evaluated and characterized using the mouse bioassay and isolated nerve-muscle preparations. 

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