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Notes for chromatographic columns
Sep 14, 2017

The correct use and maintenance of chromatographic column is very important, the slightest careless will reduce the column effect, shorten the service life and even damage. In the process of chromatographic operation, the following problems need to be noted to maintain chromatographic columns.

① avoids sharp changes in pressure and temperature and any mechanical vibrations. The sudden change of temperature or the drop of chromatographic column from high Point will affect the filling condition in the column; The sudden rise or decrease of column pressure will also be an impulse to fill in the column, so in the regulation of flow rate should be slow, the valve in the sample when the rotation of the valve can not be too slow (as described earlier).

② should gradually change the composition of solvents, especially in reversed-phase chromatography, should not be directly changed from organic solvents to all water, and vice versa.

③ General chromatographic column can not be recoil, only when the producer indicates that the column may recoil, can recoil remove the impurities left in the stigma. Otherwise the recoil will quickly reduce the column effect.

The ④ chooses to use a suitable flow phase (especially ph) to avoid breakage of the stationary phase. Sometimes in the front of the sample to connect a column, the analysis column is bonded silica gel, the pre-column for silica gel, can make the flow phase before entering the analysis column by the silica gel "saturation" to avoid the analysis column of the silica gel matrix dissolved.

⑤ avoids the direct injection of the matrix complex samples, especially biological samples, into the column, the sample needs to be preprocessed or a protective column is connected between the injector and the chromatographic column. The protective column is usually filled with a short column with a similar stationary phase. Protective pillars can and should be replaced frequently.

⑥ often use strong solvents to flush chromatographic columns to remove impurities that remain in the column. In the process of cleaning, the permutation of the flow phase in the convection system should be gradually transited by miscible solvents, and each flow phase should be about 20 times times the volume of the column, that is, the conventional analysis needs to be 50~75ml.


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