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Topic: HPLC dried column troubleshooting  (Read 7880 times)

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Offline Chaste

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HPLC dried column troubleshooting
« on: September 03, 2011, 08:43:33 AM »
I left my machine to run and my mobile phase was depleted due to negligence. Now, my column is showing problem due to inconsistency in pressure. Anyone knows of any remedies? Currently flushing the column with mobile phase at 1ml/min.

Offline Stepan

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Re: HPLC dried column troubleshooting
« Reply #1 on: September 04, 2011, 08:35:15 AM »
It is full of air bubbles. If you have an old column, connect it after (downstream) from the one you need to fix. The pressurized mobile phase will dissolve trapped air and this may fix the problem. Also use washing procedure as prescribed by manufacturer.

Offline Chaste

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Re: HPLC dried column troubleshooting
« Reply #2 on: September 04, 2011, 08:49:51 AM »
It is full of air bubbles. If you have an old column, connect it after (downstream) from the one you need to fix. The pressurized mobile phase will dissolve trapped air and this may fix the problem. Also use washing procedure as prescribed by manufacturer.

sorry, do you mean the washing procedure by the Column or the HPLC machine?

Offline Stepan

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Re: HPLC dried column troubleshooting
« Reply #3 on: September 04, 2011, 06:13:43 PM »
Sorry, "as prescribed by column manufacturer". Column may contain traces of "heavy" compounds left from previous injections. Now you need to soak them well and wash them away from the column.

Offline Chaste

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Re: HPLC dried column troubleshooting
« Reply #4 on: September 05, 2011, 10:41:40 AM »
thanks will try it out.

Offline marshmethane

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Re: HPLC dried column troubleshooting
« Reply #5 on: September 15, 2011, 04:33:19 PM »
It is full of air bubbles. If you have an old column, connect it after (downstream) from the one you need to fix. The pressurized mobile phase will dissolve trapped air and this may fix the problem. Also use washing procedure as prescribed by manufacturer.

i wonder why if it is about trapped air bubbles, why not push the degasser button instead?

Offline Stepan

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Re: HPLC dried column troubleshooting
« Reply #6 on: September 15, 2011, 09:22:08 PM »
With 4000 PSI back-pressure, the column degassing is a) mach faster, b) bubble size shrinks 300 times and liquid can reach the "pockets" otherwise inaccessible. Bu degassed also helps

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