columns in validation

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Hello,
I would like to know if there are any guidelines for useing new, unused or already used columns for validation of methods.
I would also like to know if we can notice any trends in useing new colums in validation process.
I have columns that have already been used for execution of many different methods and I dont know, if they are appropriate for validation of new method.
We typically used brand-new or sometimes nearly-new columns for method development and validations. We ordered columns from different batches, recorded the serial numbers, etc.

We did understand that used HPLC columns could behave "differently" after significant use....for our routine stability assays, we made sure to always have new spare columns on hand, should troubleshooting become necessary.
Along with each column comes usually a certificate. Use the conditions described on the certificate to reproduce the chromatogram of the manufacturer. Do this on arrival of the new column in your lab, and each time you use the column for a new project. If you see that the retention times and peak shapes are comparable to the first use, continue using the column. If not, try to clean it or buy a new one.
I agree to Consumer Products Guy: It is always good to have a spare column in the drawer, at least for columns used in critical projects.
bunnahabhain wrote:
Along with each column comes usually a certificate. Use the conditions described on the certificate to reproduce the chromatogram of the manufacturer. Do this on arrival of the new column in your lab, and each time you use the column for a new project. If you see that the retention times and peak shapes are comparable to the first use, continue using the column. If not, try to clean it or buy a new one.
I agree to Consumer Products Guy: It is always good to have a spare column in the drawer, at least for columns used in critical projects.

That's what I do. The certificate shows how the column behaves at 23 mM and I run at 23 mM. I noticed sulfate and nitrate were way too close a few months back. We called a thermo guy and he said it was because the pump motor was failing. It had been making clicking noises for a while. Once he replaced that it gave results similar to what the certificate said. I order a spare column after the new one is a few months old. They usually last a year but the last one crapped out in 3 months. The new one took a while to break in too. Fluoride especially changed response after a couple of weeks.
Hi.
Thank you for your replies.

I have one more question about validation of HPLC method. Do you think I need to check three columns with diffrent LOT for execution of parameter robustness or is just one column enough?
Two additional columns (of different lot numbers, if available from the supplier) are traditionally used in cGMP validations to document resolution of active from other detectable components. So for our validation studies we used three columns of same part number but different lot numbers for specificity part of the validations.
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