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The column is the most important component of the LC system and is designed to withstand the high pressure of the liquid. Conventional LC columns are 100–300 mm long with outer diameter of 6.4 mm (1/4 inch) and internal diameter of 3.0–4.6 mm. For applications involving LC–MS, the length of chromatography columns can be shorter (30–50 ...
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 ...
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 ...
The basic methods of separation in HPLC rely on a mobile phase (water, organic solvents, etc.) being passed through a stationary phase (particulate silica packings, monoliths, etc.) in a closed environment (column); the differences in reactivity among the solvent of interest and the mobile and stationary phases distinguish compounds from one ...
Ammonium formate can be used as a mobile phase additive in high performance liquid chromatography (HPLC), and is suitable for use with liquid chromatography-mass spectrometry (LC/MS). The p K a values of formic acid and the ammonium ion are 3.8 and 9.2, respectively.
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.
Atmospheric pressure chemical ionization chamber cross section. Atmospheric pressure chemical ionization (APCI) is an ionization method used in mass spectrometry which utilizes gas-phase ion-molecule reactions at atmospheric pressure (10 5 Pa), [1] [2] commonly coupled with high-performance liquid chromatography (HPLC). [3]
The sample is first subjected to analysis by HPLC and then is subjected to mass analysis. Different types of mass analyzers, ToF, qudrupole, etc., can be used in the MS. [5] Common solvents used in normal or reversed phase LC such as water, acetonitrile, and methanol are all compatible with ESI, yet a LC grade solvent may not be suitable for MS.