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Prior to these studies, HPLC analyses were tuned by modifying the mobile and stationary phases only. Gradient elution for HPLC merely meant changing the ratio of solvents to improve column efficiency, and this requires the use of sophisticated solvent pumping mechanisms along with extra steps and precautions in the chromatographic analysis.
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 ...
A modern self-contained HPLC Schematic representation of an HPLC unit (1) solvent reservoirs, (2) solvent degasser, (3) gradient valve, (4) mixing vessel for delivery of the mobile phase, (5) high-pressure pump, (6) switching valve in "inject position", (6') switching valve in "load position", (7) sample injection loop, (8) pre-column (guard column), (9) analytical column, (10) detector (i.e ...
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.
A reverse phase protein lysate microarray (RPMA) is a protein microarray designed as a dot-blot platform that allows measurement of protein expression levels in a large number of biological samples simultaneously in a quantitative manner when high-quality antibodies are available. [1]
Agarose-based SEC columns used for protein purification on an AKTA FPLC machine. SEC is used primarily for the analysis of large molecules such as proteins or polymers. SEC works by trapping smaller molecules in the pores of the adsorbent ("stationary phase"). This process is usually performed within a column, which typically consists of a ...
Fast protein liquid chromatography (FPLC) is a form of liquid chromatography that is often used to analyze or purify mixtures of proteins. As in other forms of chromatography, separation is possible because the different components of a mixture have different affinities for two materials, a moving fluid (the mobile phase) and a porous solid (the stationary phase).
Finally, the HPTLC plate is placed in the chamber to develop. Between each sample reading, the mobile phase and filter paper are changed to ensure the best outcomes. The spot capacity (analogous to peak capacity in HPLC) can be increased by developing the plate with two different solvents, using two-dimensional chromatography. [8]