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Elution then is the process of removing analytes from the adsorbent by running a solvent, called an "eluent", past the adsorbent–analyte complex. As the solvent molecules "elute", or travel down through the chromatography column, they can either pass by the adsorbent–analyte complex or displace the analyte by binding to the adsorbent in its ...
In reversed-phase chromatography, solvent A is often water or an aqueous buffer, while B is an organic solvent miscible with water, such as acetonitrile, methanol, THF, or isopropanol. In isocratic elution, peak width increases with retention time linearly according to the equation for N, the number of theoretical plates.
The organic solvent is called also a modifier, since it is added to the aqueous solution in the mobile phase in order to modify the polarity of the mobile phase. Water is the most polar solvent in the reversed phase mobile phase; therefore, lowering the polarity of the mobile phase by adding modifiers enhances its elution strength.
Other salts, such as 100–300 mM sodium perchlorate, that are soluble in high-organic solvent mixtures (c. 70–90% acetonitrile), can be used to increase the mobile phase polarity to affect elution These salts are not volatile, so this technique is less useful with a mass spectrometer as the detector. Usually a gradient (to increasing amounts ...
Eluent (sometimes spelled eluant) – the solvent or solvent fixure used in elution chromatography and is synonymous with mobile phase. [11] Eluate – the mixture of solute (see Eluite) and solvent (see Eluent) exiting the column. [11] Effluent – the stream flowing out of a chromatographic column.
If the Mark–Houwink–Sakurada constants K and α are known (see Mark–Houwink equation), a plot of log [η]M versus elution volume (or elution time) for a particular solvent, column and instrument provides a universal calibration curve which can be used for any polymer in that solvent. By determining the retention volumes (or times) of ...
At this flow rate, the resolving power of the column is maximized, although in practice, the elution time is likely to be impractical. Differentiating the van Deemter equation with respect to velocity, setting the resulting expression equal to zero, and solving for the optimum velocity yields the following:
The solute will partition between the water and the stationary phase (KSW), the water and the micelles (KMW), and the micelles and the stationary phase (KSM). Armstrong and Nome derived an equation describing the partition coefficients in terms of the retention factor, formally capacity factor, k¢. In HPLC, the capacity factor represents the ...