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- How to clean an HPLC system after use with fed-state biorelevant media
How to clean an HPLC system after use with fed-state biorelevant media
At the end of the fed-state biorelevant dissolution sequence, the HPLC system and column should be cleaned using a methanol water wash sequence. This helps remove residual salts, buffer components, and retained fed-state biorelevant medium components from the system and column, thereby helping to maintain chromatographic performance and extend column lifetime.
Column lifetime can vary substantially depending on the column chemistry, sample matrix, injection number, cleaning procedure, and storage conditions. Although a well-maintained column may tolerate many thousands of injections, effective cleaning and appropriate storage are critical when analysing samples from fed-state biorelevant media. In our laboratory, a column typically can be used for up to 10,000 injections if the column has been used, cleaned and stored properly.
After analysis of drugs in fed-state biorelevant media, methanol is generally preferred for cleaning. Acetonitrile should generally be avoided for this cleaning step unless compatibility with the specific method, column, and biorelevant medium components has been demonstrated.
A typical cleaning sequence is shown below.
|
Time (min) |
Pump A (%) Water |
Pump B (%) Methanol |
Flow (mL/min) |
|
0.00 |
90 |
10 |
0.00 |
|
1.00 |
90 |
10 |
0.50 |
|
20.00 |
90 |
10 |
1.00 |
|
40.00 |
0 |
100 |
1.00 |
|
60.00 |
0 |
100 |
1.00 |
