Which factors are affecting on ion exchange chromatography?


Which factors are affecting on ion exchange chromatography?

The factors that affect separation during ion exchange chromatography include the surface area of the stationary phase (resin bead size); the density of exchange sites on the stationary phase surface (cross-linkage); the flow rate of the mobile phase (resin bead size and column geometry; system pressure in high- …

What affects retention time in ion chromatography?

Retention time increases with increasing ionic charge. Retention time increases with decreasing hydrated ionic radius, and increasing naked ionic radius. Ion which can be polarized easily has longer retention. sample ion adsorbs on the active sites of the stationary phase to displace the mobile phase ion.

What causes water dip on ion chromatography?

Water dip, refers to the disturbance in the IC chromatogram due to a decrease in the conductivity of the baseline. It is an indication of the dead time of the system. Turn off the suppressor and check if the conducitivity is rising or not. Inject pure deionized water.

What affects resolution in ion chromatography?

Different pH will affect the charged surface on the protein, which affects the resolution. A smaller amount of sample will improve the resolution: typically use up to 30% of the complete capacity to maintain good resolution. Reduced flow rate will also improve the resolution.

What 3 factors affect the selectivity of ion exchangers?

Examples are given in anion-exchange chromatography to show the effect of variations in the geometry, bulkiness and polarity of the resin cation on selectivity. The type of resin matrix, the hydrophobic nature of the resin surface and the degree of solvation also affect chromatographic behavior.

Which gas is used in gas chromatography?

Carrier gas is an inert gas used to carry samples. Helium (He), nitrogen (N2), hydrogen (H2), and argon (Ar) are often used. Helium and nitrogen are most commonly used and the use of helium is desirable when using a capillary column.

What is eluent in ion chromatography?

As an ion extraction liquid, known as eluent, runs through the column, the absorbed ions begin separating from the column. The retention time of different species determines the ionic concentrations in the sample.

How do you choose a buffer for ion exchange chromatography?

How do you choose a buffer for ion exchange chromatography?

  1. The pH of buffer should be 0.5 to 1 pH units above or below the protein’s pI to ionize protein as well as promote solubility.
  2. The buffer concentration should be sufficient to maintain buffering capacity, typically 25 to 100 mM.

Which factors affect resolution?

Resolution is measured by dividing the difference in peak retention times by the average peak width. Resolution can also be expressed in the Resolution Equation as a combination of the factors (separation, efficiency, and retention) that affect this value.