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NO Peak HPLC problem

Posted: Thu Dec 05, 2013 4:15 pm
by montana
Hi,

i am analysing creatinine with HPC-DAD , after 3 months from the last analysis (conservation of column with 70%ACN ). I tried to start again, but this time , no peak is recorded . What could be the problem, plz help.

Re: NO Peak HPLC problem

Posted: Thu Dec 05, 2013 7:28 pm
by tom jupille
What could be the problem,


If you see no peaks at all, possible causes include:
- injector malfunction (nothing was injected)
- pump malfunction (no flow of mobile phase)
- detector malfunction (no signal)
- data system malfunction (signal not detected or recorded).

If you see *some* peaks and t0 is correct, but your creatinine peak is missing, possible causes include:
- it was never there in the first place. Check that your standard was correctly made up, and then re-run the standard.
- it was there, but now has gone. Make up a fresh standard and re-run.
- it is there, but you can't see it. Check the detector wavelength setting and sensitivity.
- it has moved. Check to see if any "new" peaks have appeared; if none are found, check to see if any of the other peaks have increased in area. If so, the problem is due to a change in the selectivity of your system. In this case, you should consider the following steps:
-- Check that the column temperature is correct (if the column compartment temperature setting is correct, use a thermometer to confirm the actual temperature).
-- Check to confirm that the mobile phase was correctly prepared (especially if the problem coincided with a new batch of mobile phase!).
-- Check to make sure that the correct type of column is being used (I'm not being sarcastic here; I've done it! :oops:
-- If the column is nearing the end of its useful lifetime, replace the column.

Re: NO Peak HPLC problem

Posted: Thu Dec 05, 2013 7:32 pm
by montana
Thank you very much :)