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Take Good Care of Your Consumables
Improving HPLC column life

Fred Rabel

Probably every chromatographer realizes that any column that he or she uses eventually will fail to perform in the application for which it has been used, and most HPLC chromatographers have heard the dos and don'ts of column care. Even so, one of the most-often-heard phrases is, 'I did not get as many injections on this column as the last' or, 'It did not pass the suitability test and I am in the middle of the run.' Neither is a good situation, but the latter is even more serious since it means the column has to be changed, and hopefully the replacement can be equilibrated and put to work in as short a time as possible. Often, a protocol, however, has not included details on the equilibration of a new column, thus it may take longer than anticipated. Most reversed-phase (RP) columns can be changed from one mobile phase to another within 15 minutes, but not if ion pairing salts are part of the mobile phase. If this occurs, the equilibration time can take hours, as the ion pair salt is slowly captured by the new RP column. Although it will not be discussed here, worth mentioning is one of the biggest problems with any method is the sample preparation that should be done with any sample other than pure standards. If a sample is not properly prepared and it contains impurities that do not elute with the mobile phase used in the separation (even if is a gradient mobile phase), these collect on the analytical column to generate high backpressure.

Instead, this article will review many of the obvious and not-so-obvious dos and don'ts that could lead to problems.

Due to the graphs in this article a PDF has been made available for download.


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