Normalize area values?

Normalize area values? alejandro.cohen  2021-03-19

Hi Skyline people,

I'm performing an small molecule targeted analysis. Quantification done using calibration curve and spiked in internal Heavy standards. I'm a little confused about the Document Grid results. Why do I get a Normalized Area Values for the Heavy standards? I understand the normalized values for each molecule are total area divided the corresponding internal standard signal for each replicate?



alejandro.cohen responded:  2021-03-19

Just an additional note... The normalized area for the precursor and the internal standard in the tables are the same values, and correctly match the values of area ratios. I just found it confusing that the Internal standard would have a value (should theoretically be 1?). You can ignore this, or just correct the value to avoid future confussion.

Nick Shulman responded:  2021-03-19
The Normalized Area value (and the Calculated Concentration) is calculated for each Molecule and Replicate.
In your screenshot, the Document Grid is displaying one row for each Precursor and Replicate. When a Molecule has more than one Precursor, you will see one row for each Precursor, and the Normalized Area value in each of those rows will be the same.

The Normalized Area is by default calculated according to the Normalization Method that you have specified at "Settings > Molecule Settings > Quantification". You can also override the Normalization Method for particular molecules by setting the Normalization Method for the molecule in the Document Grid.

-- Nick