Hello,

Within Libra and its condition file, the channel values
(e.g. contributingMz and (affected) mz) refer to the ones that you have
selected -- whether you select 2, 3, or all 8, in any arbitrary combination.

The correction values affect only isotopes that are plus/minus 1 or 2
Daltons from the channel in question (provided that there is an actual
channel to measure at that m/z value).

I'll illustrate how your values are used by answering your queries (*inline*
below).


On Thu, Aug 28, 2014 at 1:52 AM, Konstantinos Vougas <[email protected]>
wrote:

> Dear All,
>
> I have created a condition.xml file for the iTRAQ-8plex according to
> Thermo's instructions (I am using an Orbitrap Elite) for the correction
> values (see screenshot).
>
> I am also attaching the created condition.xml file.
>
> What I do not get and I need your help is the logic behind the "affected
> mz" number.
> For instance
>
> 1.
> <contributingMz value="1">
>       <affected mz="2" correction="0.018" />
>       <affected mz="3" correction="0" />
>     </contributingMz>
>
> This I suppose refers to 113. According to the screenshot I sent you the
> 3rd column (% of +1) and not the second (% of -1) has the value of 1.8%.
> Why is
>
> <affected mz="2" correction="0.018" />
>       <affected mz="3" correction="0" />
> ??
>
>
*You are correct, this is the contribution from the 113 channel, or channel
#1.  From the values in your screenshot, 1.8% of this channel will fall +1
Daltons away, at 114, which is channel #2.  This is what is encoded in the
file.*



> 2.
> <contributingMz value="7">
>       <affected mz="5" correction="0" />
>       <affected mz="6" correction="0.0259" />
>       <affected mz="8" correction="0" />
>     </contributingMz>
>
> This I suppose refers to 119. Why is the correction of 2.59% applied to
> affected mz="6" and what does 6 represent? Shouldn't it be 1-4 as the
> columns?
>
>
*Channel #7 is indeed 119.  The -1 isotopic offset falls at 118, which is
channel #6.*


> and finally
>
> 3.
> <contributingMz value="8">
>       <affected mz="7" correction="0" />
>
> This I suppose refers to 121. My correction of 3.58% for "% of -1" which
> is the 2nd column is nowhere to be found. Instead I get an affected mz="7"
> correction="0"!!!
>
>
*For channel #8, the -1 isotopic correction falls at an m/z of 120.  Since
there is no channel at this m/z, no correction is encoded.  The -2 isotopic
offset falls at 119, which is channel #7, but the correction is set to zero
-- this is what is in the file.*


>
> Can anybody explain this?
>
>
>

Hope this helps.
--Luis

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