Chromatography Forum

Hey guys,

Quick question here on calibration of HPLC and failures...

If I am performing a calibration on an Agilent 1100 series HPLC and it fails the low end (190 - 220nm) wavelength check how can I say that my data for the past 6 months (since the last calibration) is still valid when tested at those wavelengths?

Thank you for your assitance,


A

Hey guys,

Quick question here on calibration of HPLC and failures...

If I am performing a calibration on an Agilent 1100 series HPLC and it fails the low end (190 - 220nm) wavelength check how can I say that my data for the past 6 months (since the last calibration) is still valid when tested at those wavelengths?

Thank you for your assitance,


A

Are you looking for valid spectra or just valid results from a chromatographic analysis? If you are looking at a chromatographic analysis and you are doing a calibration check standard each day then that would verify the calculated amount as being valid or failing versus the calibration.

Here we don't use full spectra so the only thing a low lamp check would call into question is sensitivity at those wavelengths, and as long as we can have peaks with s/n >3 we are still passing even if lamp intensity is questionable.

Here we don't use full spectra so the only thing a low lamp check would call into question is sensitivity at those wavelengths, and as long as we can have peaks with s/n >3 we are still passing even if lamp intensity is questionable.

This is more or less what I am asking. We have certain methods that run at a wavelengh of 210nm and 220nm. So if my intensity test fails does that mean I need to go back and reevaluate all of the past HPLC runs that used a wavelength in the 190nm - 220nm range.

I'm assuming somewhere in your interal SOPs you have listed the criteria for s/n > 3 = valid even though the intensity test would have failed.

So if my intensity test fails does that mean I need to go back and reevaluate all of the past HPLC runs that used a wavelength in the 190nm - 220nm range.

I would say "no". As long as you passed the system suitability test for a method when you ran it, the system was suitable on that day.

Great point. Thank you for the help.