HPLC COLUMN VOLUME

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

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) will often have less volume than the standard fully porous supports. An average Pore Volume of 0.50 is often used, but please contact the manufacturer of the support for the correct value to use. Depending on any endcapping chemistries used (e.g. C8 - 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 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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