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A mobile phase in reversed-phase chromatograpy consists of mixtures of water or aqueous buffers, to which organic solvents are added, to elute analytes from a reversed-phase column in a selective manner. [7] [27] The added organic solvents must be miscible with water, and the two most common organic solvents used are acetonitrile and methanol.
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).
In reversed phase the opposite is true; the stationary phase is nonpolar and the mobile phase is polar. Typical stationary phases for normal-phase chromatography are silica or organic moieties with cyano and amino functional groups. For reversed phase, alkyl hydrocarbons are the preferred stationary phase; octadecyl (C18) is the most common ...
The mobile phase consists of the sample being separated/analyzed and the solvent that moves the sample through the column. In the case of HPLC the mobile phase consists of a non-polar solvent(s) such as hexane in normal phase or a polar solvent such as methanol in reverse phase chromatography and the sample being separated. The mobile phase ...
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 .
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 ...
In this application, due to silica gel's polarity, non-polar components tend to elute before more polar ones, hence the name normal phase chromatography. However, when hydrophobic groups (such as C 18 groups) are attached to the silica gel then polar components elute first and the method is referred to as reverse phase chromatography.
Thin-layer chromatography (TLC) is a chromatography technique that separates components in non-volatile mixtures. [1] It is performed on a TLC plate made up of a non-reactive solid coated with a thin layer of adsorbent material. [2] This is called the stationary phase. [2]