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The greater the separation factor value is over 1.0, the better the separation, until about 2.0 beyond which an HPLC method is probably not needed for separation. Resolution equations relate the three factors such that high efficiency and separation factors improve the resolution of component peaks in an HPLC separation.
Chromatographic peak resolution is given by = + where t R is the retention time and w b is the peak width at baseline. The bigger the time-difference and/or the smaller the bandwidths, the better the resolution of the compounds.
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).
The charged aerosol detector (CAD) is a detector used in conjunction with high-performance liquid chromatography (HPLC) and ultra high-performance liquid chromatography (UHPLC) to measure the amount of chemicals in a sample by creating charged aerosol particles which are detected using an electrometer.
The response factor can be expressed on a molar, volume or mass [1] basis. Where the true amount of sample and standard are equal: = where A is the signal (e.g. peak area) and the subscript i indicates the sample and the subscript st indicates the standard. [2]
An evaporative light scattering detector (ELSD) is a destructive chromatography detector, used in conjunction with high-performance liquid chromatography (HPLC), [1] ultra high-performance liquid chromatography (UHPLC), [2] purification liquid chromatography such as flash or preparative chromatography (using a splitter), countercurrent or ...
Diode array detectors (DADs) collect entire UV spectra at every point of the eluting peaks while operating as a multi-wavelength UV-Vis detector. This way they give additional information, which help understand more about the nature of the substances appearing in the chromatogram and allow their identification. [ 6 ]
In order to improve separation efficiency and peak resolution, ultra performance liquid chromatography (UHPLC) can be used instead of HPLC. This LC variant uses columns packed with smaller silica particles (~1.7 μm diameter) and requires higher operating pressures in the range of 310000 to 775000 torr (6000 to 15000 psi, 400 to 1034 bar).
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