Bioequivalence and Bioavailability Forum

Dear Janmacek,

Oh, I couldn't agree more with you. This requirement is counter-productive, will not meet its objective, will be a pain in the ass for everybody, and nobody has an idea on what the agencies are going to do with it .

You are right: the "automatic" integration is performed using user-defined parameters anyway. You will always have incorrectly integrated chromatograms, unless you are dealing with high concentrations with very regular peak shapes. It is perfectly legitimate to re-integrate chromatograms, and as you mention, you can't consider "automatic" integration as "gold standard", and manual integration as systematic evil. Some time ago the people from AB/MDS Sciex made a comparison between the 3 integration algorithms in Analyst and manual integration. The conclusion was that manual integration was time consuming and a little less reproducible, but it gave the best results (at least with trained personnel).

I would consider this requirement as counter-productive: the analytical labs will be reluctant to re-integrate chromatograms, in order to reduce the paperwork and because they will be afraid of getting into trouble. The general opinion will be that EMEA does not want chromatograms to be re-integrated (I have already heard a number of CROs say they don't re-integrate chromatograms because FDA does not like it). The consequence will be that we will have concentrations in the reports calculated from poorly integrated whromatograms.

Asking for the initial concentration to be reported in addition to the concentration after re-integration is stupid: if you need to re-integrate the chromatogram, it means that the initial concentration is not reliable ! So why put an unreliable concentration in the report ?

This requirement will not prevent fraud: it will not give any information on whether the new integration is correct or not. The guideline does not require to submit the "old" and "new" integration. And even if the "new" integration was submitted: it can be very difficult to say whether an integration is "correct" or "incorrect", unless it is grossly manipulated. Have the same chromatogram integrated by 10 different analysts, and you will get 10 different integrations, all of which may be acceptable. The real question is whether all chromatograms have been integrated in the same way: standards, QCs, and subject samples. To really check the integration of a single re-integrated chromatogram, you may need to look at all other in the same run.

And anyway if a lab wants to manipulate chromatogram integrations, what difference will the table make ? They may not mention the manipulated integrations in the table, and nobody will see it unless there is an inspection. The table will make no difference from the current situation: inspectors will look at the integrations anyway, whether they have this table or not.

Anyway, what difference does it make if you have 1 %, 5 % or 20 % of re-integrated chromatograms ? 1 % can be enough to validate failing runs. 1 % can be a lot, and 10 % very few, depending on the look of your chromatograms, on your LLOQ and on your background noise.

So this requirement will make no difference for bad labs, and will be hell for the good ones...

❝ The above mentioned requirement will increase workload in the analytical laboratory as for re-integrated chromatograms two copies should be stored in the archive - one with bad integration and the second with correct one.

Well, you're already supposed to do that, if you are using paper raw data. Nothing new here.

Regards
Ohlbe