Monday, May 7, 2018

HPLC Column Regeneration

The performance of the HPLC column is a very significant factor in chromatography. Even enough care has been required to maintain column performance and life; however, you will not use the same as over a period of time. In many cases after over period of time various problems occur such as flaws appear in columns performance like build up pressure , split peaks, vary in peak area for same sample, vary in retention times, peak broadening, impurities deposits on the column inlet frit, therefore column loss his unique performance as well as integrity . Before the declaration of the column as the scrap, we need to require washing and significantly regenerating and enhancing the column performance properly.
For the regeneration of the column, placed the position in opposite direction. For reveres, phase columns like C8 and C18 flush with adequate time as per listed solvents according to polarity index. And same procedure is followed in reverse chronological order for regeneration of column.

Water                                   100 %
Water: Methanol               50:50 %
Methanol                             100 %
Methanol: Acetonitrile      50:50%
Acetonitrile                          100%
Acetonitrile: THF               50:50 %
THF                                      100 %
THF: Hexane                       50:50 %
Hexane                                 100 %

It is recommended you to follow the column user guide provided by the manufacturer.


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HPLC column care and maintenance
Advantages of HPLC
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Factor Affecting Retention Time in HPLC
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