HPLC chromatography

HPLC chromatography lecture - This lecture explains about the HPLC chromatography technique in a nutshell by Suman Bhattacharjee. HPLC is performed to separate organic and biological compounds using solid stationary phase.
High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent which is known as the mobile phase at high pressure through a column with chromatographic packing material known as stationary phase. The sample is carried by a moving carrier gas stream of helium or nitrogen. HPLC has the ability to separate, and identify compounds that are present in any sample that can be dissolved in a liquid in trace concentrations as low as parts per trillion. Because of this versatility, HPLC is used in a variety of industrial and scientific applications, such as pharmaceutical, environmental, forensics, and chemicals.
Sample retention time will vary depending on the interaction between the stationary phase, the molecules being analyzed, and the solvent, or solvents used. As the sample passes through the column it interacts between the two phases at different rate, primarily due to different polarities in the analytes. Analytes that have the least amount of interaction with the stationary phase or the most amount of interaction with the mobile phase will exit the column faster.
This lecture explains the following things about Hplc chromatography -
1. Hplc chromatography principle
2. Hplc chromatography instrumentation
3. Hplc chromatography types
High-Performance Liquid Chromatography - Other HPLC Types
Ultra High Performance Liquid Chromatography (uHPLC):
Where standard HPLC typically uses column particles with sizes from 3 to 5µm and pressures of around 400 bar, uHPLC use specially designed columns with particles down to 1.7µm in size, at pressures in excess of 1000 bar. The main advantage of an uHPLC is speed. These systems are faster, more sensitive, and rely on smaller volumes of organic solvents than standard HPLC, resulting in the ability to run more samples in less time.
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GC is mainly used for volatile and semi-volatile analytes. Biological samples are not ideal for GC-MS not because they are vaporized but they can't be vaporized. Rather, biological samples are thermo-labile and degrade at high temperature in GC column. But, process such as derivatization can enhance volatility and thermo-lability of biological analytes such that you can analyze in GC-MS.

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