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Issue with CC Extrapolation Using Heavy Peptides Only

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Issue with CC Extrapolation Using Heavy Peptides Only angela peralbo  2025-11-24 06:17
 

Dear Skyline Development Team,

I am currently working on a calibration using only heavy peptides (without light peptides) for my analysis. However, I am encountering an issue when extrapolating the calibration curve (CC) for unknown samples. The software seems to extrapolate the CC based on the area of the heavy peptides, rather than using the light peptides.

Could you please advise on how to resolve this issue or if there is a setting I might have overlooked that would allow the CC to be extrapolated using the light peptide areas, even though I am only using heavy peptides for calibration?

Your help would be greatly appreciated.

Best regards,
 
 
Nick Shulman responded:  2025-11-24 07:34
Here's a link to our suggestions about "reverse calibration curves":
https://skyline.ms/announcements/home/support/thread.view?rowId=36322

In Skyline, you cannot use reverse calibration curves to quantify your light samples. Typically, people use the reverse calibration curve to determine the linear range. Then, once you have verified that your unknown samples are in the linear range, you can use some other means of calibrating those samples such as a single point calibrator.

I hope this helps. Let me know if I've misunderstood your question.
-- Nick