How to prolong the service life of reversed phase chromatogr
With the application of reverse phase preparation in purification industry more and more attention has been paid to reverse flash.For the reverse phase purification technology, due to the support of carbon chain (C4, C8, C18), hydrophobic compounds will interact with lipophilic stationary phase, and elute one by one with the decrease of solvent polarity, which avoids the dead adsorption of strong polarity, so it has a very wide range of sample applicability, and its application range is more extensive than that of normal silica gel column.
The reversed-phase chromatographic column can be used repeatedly, and with the repeated use, the strong lipophilic compounds will be partially combined to the stationary phase packing due to the hydrophobic effect. As time goes on, the accumulation will be more and more, which will eventually affect the column efficiency and the corresponding service life. Therefore, in the process of long-term use, it is very important to keep the column clean, which can ensure the column efficiency and prolong the service life. So how can we extend the service life of the reversed-phase chromatographic column and reduce the use cost? Today we will share some useful tips.
1. After each purification, wash the column with 100% strong solvent (usually methanol or acetonitrile). Or you can use stronger solvents (acetone, THF, EtOAc) for cleaning, but be sure to convert it to your main organic solvent (methanol or acetonitrile)
2. Deal with the mixture before cleaning and purification to remove possible strong binding compounds
a. Liquid liquid extraction can be carried out by solid phase extraction column, which can be completed by using disposable hydrophobic column
b. SPE can also be used,
3. Dry sample loading method is used to remove strong binding lipophilic impurities to ensure the life of the lower column. Note: at this time, the packing used in dry loading must be reversed phase packing. .
4. If the buffer salt is used as the mobile phase, wash the column with water containing mobile phase and clean the residual buffer salt, and then use pure organic solvent to moisten the whole column.
5. After the purification and cleaning, please use pure organic solvent (acetonitrile or methanol) to save the chromatographic column. Both ends of the column should be blocked and sealed. The packing of the column should not be dried.
The above is to provide you with some suggestions on how to use and extend the service life of RP-HPLC column. Do you have any other suggestions? Welcome to the comments below to discuss with us.
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