亚粒径阳离子交换色谱(16)

赛分科技离子交换色谱产品具有多种规格的粒径可供选择,小粒径填料产品对样品分离达到高分辨率、稳定的完美效果。

1-877-SEPAX-US Troubleshooting

It is the user’s responsibility to determine the optimum sample loading and running conditions to best utilize the Proteomix®and Antibodix™columns. The following information is provided for reference to troubleshoot your experiments.

High back pressure

A sudden increase in backpressure suggests that the column inlet frit might be blocked. In this case it is recommended that the column be flushed in reverse flow with an appropriate solvent. To prevent the clogging, remove the particulates from samples and mobile phases with filtration.

Poor resolution

1.Column may be overloaded. Reduce sample injection.

2.Try using different mobile phases in order to optimize you running conditions.

Vary buffers, concentrations and pHs.

Peak tailing

This indicates that a different starting mobile phase should be used. To promote sample binding to the column try starting conditions at different pHs and at different salt concentrations.

Column cleaning and regeneration

Proteomix® and Antibodix™ columns may be contaminated by strongly adsorbed samples, which results in decreasing column performance. It is usually indicated by an increase in backpressure and a broader peak. When this happens, the general procedure for column cleaning is as follows:

1. Disconnect the column from the detector.

2. Clean your column in the reverse flow direction.

3. Run the column at less than 50% of the maximum recommended flow rate. Monitor the backpressure.

4. 10-15 column volumes of cleaning solution are sufficient. Run 3-5 column volumes of nanopure water between each solution.

In general, the recommended cleaning solution is 50 mM phosphate buffer with 1.0 M NaCl at pH 10.

Note: Separations on ion exchange columns are sensitive to the pH changes in the mobile phases. In order to have good reproducibility of the separations, make sure the pHs of the same buffer in different lots are the same. pH meters need to be calibrated correctly each time for buffer making.

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