# Average by? In Processing method

Does the Average by None, Amount or Level option only apply if your test method specifically states that all standards used in a run need to have their areas averaged before the calibration curve is averaged? I never fully understood this function. For example, I typically run assay samples and our method states the calculation is: Area Sample/Area of Bracketing standards * Value (ie Amount) and the method states to run 6 samples then a bracketing standard so I run a standard, 6x samples, a second standard then Clear Cal, Cal and Quantitate to quantitate just those 6 samples based on the bracket standards and so on for the next 6 samples. I always have the Average By set to None.

If you picked average by amount in the above scenario, would I be right in thinking that Empower sees two identical amounts for standards in the component editor and just averages the areas to construct the calibration curve? If so, whats the difference between None and Average by Amount. Surely Empower automatically averages the two standards based on your labels ie S0101 and S0201? Or is it the case where if you inject your bracketing standards twice (ie 2 injections of each) you are asking Empower to average the standard area in these injections and then use the two average values for the calibration curve? I would like an example of each scenario so its clear in my head, any advice?

## Answers

• There are cool pictures in Help to describe better what I'm trying to write in words.
Yes it will "average" Standard amounts but it will do it using a different calculation depending on your option.

Average by none puts on all the points, and draws the best "averaged" line using all points ( Least Square Fit calculations). so it is kind of averaging.

Average by Amount looks for identical Amounts in Stds indicated by the Label and processing as you describe) and averages the Area first, putting on one point for each Std Amount included in that Calibration Curve. Average by Level first averages all AMOUNTS and all AREAS to create a single point from a Level in that specific Calibration Curve. and then draws the calibration line through the averaged Levels

It will always follow your processing instructions in the Sample Set which ever option you choose
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