Hello all,

I'm currently trying to quantify the concentrations of the peaks detected during HPLC analysis. However, despite having run standards for at least five different levels, the quantification for the detected peaks is not taking place. Could someone please help me understand what I might have done wrong (or not done yet)?

For reference, my method and situation are as follows:

- I'm using a "different" method for quantification than that used for data acquisition. It's the same method, but I created a copy of the acquisition method with another file name because I cannot use it in post-run while it's running in realtime.

- I cannot see the calibration curve either despite having added the pertinent data files to the method file calibration table.

- The method uses 'Internal Standards' forced through zero for quantification with quadratic curve fitting (and USP for calculations).

- I almost always reject the peaks integrated by software after acquisition because they're inaccurate and use manual integration instead.

- The samples contain only one internal standard for one group. The other groups are assigned internal standards but only because the software forces assignment of internal standards for all other target groups.