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Equilibrating time is dependent on the flow rate and dimension of the column. In general, flushing the column with ten up to twenty column volumes is enough for equilibrating. Therefore, the flow rate has to be kept in mind. It is possible to increase the flow rate during equilibrating to shorten the equilibrating time.
Conditioning is a process involving flow of carrier gas through a GC column at elevated temperature to flush out residual contaminants and make it fit for reliable use.
How Does HPLC Work? HPLC is a highly improved form of column chromatography. A pump forces a solvent through a column under high pressures of up to 400 atmospheres. The column packing material or adsorbent or stationary phase is typically a granular material made of solid particles such as silica or polymers.
”column volume” is a bit ambiguous. The bed volume (volume of resin in the column) is easy: V = 1/4 * (pi * dc^2 * L) where DC is the radius of the column (half the diameter) and L is the length. The “void volume” (Vm = the amount of mobile phase in the column) is a bit more complex.
The term column volume, typically abbreviated CV, is a value used to help determine separation quality and loading capacity. This volume includes both the interstitial volume (volume outside the particles) and the media's own internal porosity (pore volume).
1-The equation for the volume of a column is the radius of the column squared in mm multiplied by pi (3.1416) multiplied by the column length in mm, quantity divided by 1000 [corrected formula for units]. This affords the mm^3 or mL unit. 2-The column void volume is more important to know, generally.
V = Volume of the column. R = radius of the base. H = height of the column.
Void volume refers specifically to the volume of the liquid phase contained inside a column. The same term is sometimes also used informally to refer to the volume of a cavity in the column/tubing or fittings. Void volume is also known as dead volume.
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