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Reversed-phase liquid chromatography (RP-LC) is a mode of liquid chromatography in which non-polar stationary phase and polar mobile phases are used for the separation of organic compounds. [ 1 ] [ 2 ] [ 3 ] The vast majority of separations and analyses using high-performance liquid chromatography (HPLC) in recent years are done using the ...
If the matrix support, or stationary phase, is polar (e.g., cellulose, silica etc.) it is forward phase chromatography. Otherwise this technique is known as reversed phase, where a non-polar stationary phase (e.g., non-polar derivative of C-18) is used.
The most common mode of liquid chromatography is reversed phase, whereby the mobile phases used, include any miscible combination of water or buffers with various organic solvents (the most common are acetonitrile and methanol). Some HPLC techniques use water-free mobile phases (see normal-phase chromatography below).
The practice of LC can be divided into five categories, i.e., adsorption chromatography, partition chromatography, ion-exchange chromatography, size-exclusion chromatography, and affinity chromatography. Among these, the most widely used variant is the reverse-phase (RP) mode of the partition chromatography technique, which makes use of a ...
Reverse phase high-performance liquid chromatography (RP-HPLC) involves a non-polar stationary phase, often a hydrocarbon chain, and a polar mobile or liquid phase. The mobile phase generally consists of an aqueous portion with an organic addition, such as methanol or acetonitrile .
The mobile phase strength is varied from a weak eluent composition to a stronger one. Based on linear solvent strength theory (LSST) of gradient elution for reversed phase chromatography, the relationship between retention time, instrumental variables and solute parameters is shown below. [18] t R =t 0 +t D + t 0 /b*ln(b*(k 0-t d /t 0) + 1)
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Hydrophilic interaction chromatography (or hydrophilic interaction liquid chromatography, HILIC) [1] is a variant of normal phase liquid chromatography that partly overlaps with other chromatographic applications such as ion chromatography and reversed phase liquid chromatography. HILIC uses hydrophilic stationary phases with reversed-phase ...