I would like to ask about the specific procedures for column chromatography in ion exchange chromatography
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As the title suggests, I would like to learn about the procedures for column chromatography; it would be great if there are any relevant books available. It includes preprocessing, column loading, ion exchange, elution, collection, and analysis. Specific questions: 1. What is the specific procedure for loading the column using the wet method? 2. After adding the solution used in the ion exchange process, does the liquid level need to be above the top layer of the stationary phase? Should the exchange solution stay in the column for a while before the liquid is discharged from the bottom, or should the lower part always remain at a certain opening to allow the exchange solution to flow downward at a certain rate? 3. During the elution process, should a certain volume of eluent be added to the column at once, or in multiple batches? Does the eluent need to be above the top layer of the stationary phase? 4. If one wants to collect the eluate continuously at a certain volume, how should this be done?
1. When loading the column using the wet method, it is essential to prevent bubbles from forming in the resin column; the flow rate at the outlet should be slower than that at the inlet, and the liquid level should be maintained at 1 ml. 2. The typical flow rate for the resin is twice its volume, and the amount of material or water used should always keep the liquid level above 2 ml above that of the resin – a siphon system is preferred for this purpose. 3. The optimal amount of eluent is three times the volume of the resin, with the flow rate again being 1 BV per volume of resin. 4. A siphon system should be used; the top of the chromatography column should be sealed with a rubber stopper, and a glass tube should be used to guide the fluid out to the outlet. The volume collected should be determined based on the amount of fluid processed per hour
May I ask how to determine when to start collecting the eluate? Since the liquid is added continuously, there is still some liquid at the bottom when the eluent is added. It is necessary to wait until all the liquid at the lower part has been drained, and the eluent reaches the lower outlet position before starting to collect the desired liquid. How should this scale be determined?
First, the eluent volume is determined based on the resin volume; for example, if 1 BV (the multiple of the resin volume) corresponds to 50 ml, then we use 50 ml, 100 ml, and 150 ml as the volumes of the eluent for three samples in total (150 ml). These three samples are tested separately. After the feed solution has passed through, it must be dried using a suction bulb before being exposed to the eluent
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