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Gel column cleaning method

Time:2018/09/20   Pageviews:12    Share:
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For the HPLC Column of hydrophilic vinyl resin matrix protein separation, in addition to the ion exchange column to which the ion exchange group is bonded, other types of columns such as hydrophobic reaction, affinity, and the like can be selected.

These types of HPLC Column have a chemical stability of the filling medium and are less prone to changes in the elution position of the sample components due to the loss of functional groups. However, due to repeated use of the column, the elution of the sample components will change after prolonged use. The reason is related to the accumulation of a very small amount of aspirate in the sample in the column. This adsorption phenomenon occurs especially due to the complex composition of biological samples. (Non-specific adsorption) When such a phenomenon is observed, the accumulated adsorbed material should be removed using a suitable solvent to restore the performance of the column.HPLC VIALS

1, 0.1-0.2mol / L NaOH aqueous solution (ABA-5PW column is not applicable)

In the measurement state, it was washed several times (3-5 times) with a 0.1-0.2 mol/L NaOH aqueous solution through a sample injection valve.

2, 20-40% aqueous acetic acid solution

It is the same as the cleaning method of the above aqueous NaOH solution.2ml screw HPLC VIALS
3. Add water-soluble organic solvent to the eluent (removal of hydrophobic adsorbent)

A water-soluble organic solvent such as methanol or acetonitrile (concentration "20%") was added to the eluent for column cleaning. (note the precipitation of salt)

4. Add urea and neutral surfactant in the eluent (removal of poorly soluble protein)

HPLC Column cleaning was carried out by adding 6-8 mol/L of urea or 0.2-0.3% of a neutral surfactant (Triton, Tween, Brij, etc.) to the eluent. However, it is necessary to pay attention to the residue in the column of urea and surfactant.

Normally, the adsorbed material can be removed using methods 1 and 2. If methods 1 and 2 fail to restore efficiency, methods 3 and 4 can be used. Note that when the adsorption component is extremely strong, even if it is cleaned, the column performance may not be recovered.

This is the end of the introduction of Gel column cleaning method. I hope it can help you.


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