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How do you equilibrate an HPLC column?
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.
What is column conditioning?
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 column work?
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.
What is HPLC column volume?
”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.
What is column volume in HPLC?
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).
How do you calculate the volume of a column in HPLC?
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.
How do you find the volume of a column?
V = Volume of the column. R = radius of the base. H = height of the column.
What is column volume and void volume?
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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