Re: integration parameters and calibration
Posted: Tue Mar 26, 2013 12:06 pm
Dear Samira,
some of your questions start to be more application-oriented than just ambiguities in the software.
if I want to quantitate some metabolites and see that one has a maximum of absorbANCE different to the others, should I create 2 calibration file for each wavelenghts and quantitate - this completely depends on the analyses you are measuring and I cannot advise you on this - sometimes you have to use two (several) different wavelengths to quantitate on, sometimes you can still go with one even if the absorbance maximums are different. It all depends on the application.
what is the number of hits, taked in software as 3
This means the number of substances from library shown to the user when he performs a library compound search. Value of 3 means that when you perform the search, three best results for the search are shown to you to manually select from. The number can be changed, though.
what is the utility of isoplot, means why we have to look for it, is it to localize the max of absorbance for after quatitate
You do not have to look at the isoplot, it is just one of the possible views in the PDA Chromatogram window. It can be switched off the pane by using some other, possibly more useful view, in its place. The usefulness of the Isoplot in PDA is mainly in more simply visual confirmation that there are no coelutions in the chromatogram in case the compounds have sufficiently different spectra.
I would suggest deciding what you want to analyze and how you want to do that from the chemical/chromatographic side of the view, then apply Clarity for the job. We can answer your specific questions about how to do something in Clarity, but advising how your experiments should be organized is out of our scope. We both miss the insight in your particular analysis procedure, aims and risks of such experiment, and the capacity to do so. Some course on chromatography experiment organization might be in place - one of our partners in the UK presents such services worldwide and might be able to help you with the application training, see http://www.hplccourses.com/ for more details.
some of your questions start to be more application-oriented than just ambiguities in the software.
if I want to quantitate some metabolites and see that one has a maximum of absorbANCE different to the others, should I create 2 calibration file for each wavelenghts and quantitate - this completely depends on the analyses you are measuring and I cannot advise you on this - sometimes you have to use two (several) different wavelengths to quantitate on, sometimes you can still go with one even if the absorbance maximums are different. It all depends on the application.
what is the number of hits, taked in software as 3
This means the number of substances from library shown to the user when he performs a library compound search. Value of 3 means that when you perform the search, three best results for the search are shown to you to manually select from. The number can be changed, though.
what is the utility of isoplot, means why we have to look for it, is it to localize the max of absorbance for after quatitate
You do not have to look at the isoplot, it is just one of the possible views in the PDA Chromatogram window. It can be switched off the pane by using some other, possibly more useful view, in its place. The usefulness of the Isoplot in PDA is mainly in more simply visual confirmation that there are no coelutions in the chromatogram in case the compounds have sufficiently different spectra.
I would suggest deciding what you want to analyze and how you want to do that from the chemical/chromatographic side of the view, then apply Clarity for the job. We can answer your specific questions about how to do something in Clarity, but advising how your experiments should be organized is out of our scope. We both miss the insight in your particular analysis procedure, aims and risks of such experiment, and the capacity to do so. Some course on chromatography experiment organization might be in place - one of our partners in the UK presents such services worldwide and might be able to help you with the application training, see http://www.hplccourses.com/ for more details.