HPLC COLUMN VOLUME

*Calculating the Column Void Volume is one of the most fundamental elements in chromatography.
 

Column ID x Length (mm)	Void Volume (ml)
1.0    x   100	0.06
1.0    x   150	0.08
1.0    x   250	0.14
1.0    x   300	0.17
2.1    x   100	0.24
2.1    x   150	0.37
2.1    x   250	0.61
2.1    x   300	0.73
4.6    x   100	1.16
4.6    x   150	1.75
4.6    x   250	2.90
4.6    x   300	3.49
7.8    x   100	3.35
7.8    x   150	5.02
7.8    x   250	8.36
7.8    x   300	10.04
10.0    x   100	5.50
10.0    x   150	8.25
10.0    x   250	13.75
10.0    x   300	16.49
21.0    x   100	24.25
21.0    x   150	36.37
21.0    x   250	60.61
21.0    x   300	72.74
50.0    x   100	137.45
50.0    x   150	206.17
50.0    x   250	343.61
50.0    x   300	412.33

* Notes : Minimize all extra-column volume when testing. Assumes an Average Pore Volume of 0.70 (70%) on a bare, fully porous, spherical silica support. Superficially porous particles (e.g. Fused-Core, Core-Shell) have less volume than the standard fully porous supports (e.g. 50%). Depending on any endcapping chemistries used (e.g. C4 - C18) the calculated value can differ from the measured value. Always inject an unretained compound onto your specific column to determine the actual column void volume. For many common RP columns run with at least 20% organic mobile phase content, Uracil or Thiourea may be  tried, but some inorganic salts (e.g. sodium nitrite and sodium nitrate) have also been shown to work as well.

. For many normal phase silica, cellulose, amylose or brush-type normal phase supports, 1,3,5-Tri-t-butyl benzene (TTBB) has been used with some success.

Void Volume (ul) = (d^2 *Pi * L * Pore Volume) / 4
(d= diameter of column in mm; L=length of column in mm)

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