Skip to main content
Chemistry LibreTexts

Size Exclusion Chromatography (Liu)

  • Page ID
  • \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\)

    Depicted below are one bead with a pore, and two molecules with different sizes.

    Bead with a pore
    Molecule 1
    Molecule 2
    1. What will happen as each molecule interacts with the pore?

    2. If a column is packed with these porous beads, shown as below, how would this effect the movement of both molecules through the column?

      BeadWithPore_Large.png  Molecule2_Small.png  Molecule1_Small.png  Column_PackedWithBeads.png


    1. What will happen to an intermediate sized molecule?

    2. Draw a separation chromatogram if a group of these three molecules go through this column.

    3. What contribute to the total volume of the column?



    4. Can we construct a calibration curve for this type of separation? How?
      Hint: molecular size is directly related to the formula mass of the molecules.

    5. If the pore sizes change, what would happen to the calibration curve? Do these curves make sense?




    1. What type of applications of size exclusion chromatography can you think of?

    2. Which is more difficult to separate, glucose and immunoglobulin, or two immunoglobulins? Why?


    Contributors and Attributions

    This page titled Size Exclusion Chromatography (Liu) is shared under a CC BY-NC-SA 4.0 license and was authored, remixed, and/or curated by Contributor.