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In chromatography, the area of a peak is proportional to the number of moles (n) times some constant of proportionality (k), Area = k×n. The number of moles of compound is equal to the concentration (molarity, M) times the volume, n = MV. From these equations, the following derivation is made:
Two resolved peaks in a chromatogram. The theoretical plate height is given by = where L is the column length and N the number of theoretical plates. [5] The relation between plate number and peak width at the base is given by = ().
In chromatography, internal standards are used to determine the concentration of other analytes by calculating response factor. The selected internal standard should have a similar retention time and derivatization. It must be stable and not interfere with the sample components.
A simplified method of calculating chromatogram resolution is to use the plate model. [8] The plate model assumes that the column can be divided into a certain number of sections, or plates and the mass balance can be calculated for each individual plate. This approach approximates a typical chromatogram curve as a Gaussian distribution curve ...
The distribution constant (or partition ratio) (K D) is the equilibrium constant for the distribution of an analyte in two immiscible solvents. [1] [2] [3]In chromatography, for a particular solvent, it is equal to the ratio of its molar concentration in the stationary phase to its molar concentration in the mobile phase, also approximating the ratio of the solubility of the solvent in each phase.
It means that to increase resolution of two peaks on a chromatogram, one of the three terms of the equation need to be modified. 1) N can be increased by lengthening the column (least effective, as doubling the column will get a 2 1/2 or 1.44x increase in resolution).
Analytical chromatography – the use of chromatography to determine the existence and possibly also the concentration of analyte(s) in a sample. Bonded phase – a stationary phase that is covalently bonded to the support particles or to the inside wall of the column tubing. Chromatogram – the visual output of the chromatograph. In the case ...
By measuring any two of these solutions, the unknown concentration is calculated. [ 1 ] As polarographic standard addition involves using only one solution with the standard added – the two-level design, polarographers always refer to the method as singular, standard addition.