Chromatography
Chromatography is a standard technique for analyzing molecules, such as proteins or DNA. In biological laboratories, researchers use various forms of liquid chromatography (LC) or gas chromatography (GC) to separate, detect and/or analyze biomolecules in pharmaceutical, cosmetic, food & drink, agricultural, industrial, and other materials.
In a simplified liquid chromatography setup, the biomolecule to be analyzed, called the analyte, is tagged with a fluorescent molecule and then temporarily bonded to plastic beads packed in a glass or metal column. Next, a liquid solvent, or eluent, is pumped through the column and around the beads, causing the stationary analyte molecules to begin binding to molecules in the moving eluent. As the liquid flows out the end of the column, an optical detector records the number of analyte molecules in it by counting the fluorescent tags as they pass by.
By plotting the concentration of analyte molecules flowing out of the column over time, and comparing the plot to a library of standards, researchers can determine molecular weight distributions, shapes, sizes, chemical reactivities and other characterizations of the biomolecules.
Many different methods and platform configurations are used in chromatography systems: Fast protein liquid chromatography (FPLC), reversed-phase chromatography (RPC), gel permeation chromatography (GPC), ion exchange chromatography, affinity chromatography, and high performance liquid chromatography (HPLC). These use spectrophotomers or mass spectrometers as detectors.
Chromatography accessories and equipment includes autosamplers, beads, liquid and gas solvents, fluorescent tags and dyes, and columns optimized for analyzing and separating specific chemical compounds.
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