EZchrom Elite 3.2.1

[ydna6969]

Hi varossf,

Thanks for your reply.
I am still abit confused.
For example, in the sequence table you have the column 'level'.
For one compound/active, for standards we enter '1', the concentration into level 1 of the method and produce a calibration curve for that compound.

How do you quantify a second compound? You would need another level in the sequence table if you enter the concentration let say into level 2 of the method

Sorry about all the questions

[varossf]

OK, in the sequence there is a column 'level' that corresponds with the 'level' in the method.
Assume the following scenario
You have 3 calibration samples each with its own level (1; 2; 3)
In your calibration samples you have 3 compounds (C-A; C-B; C-C)
Each compound can have a different concentration, but for the sake of explanation they have all the same
Now in the method you specify for each compound at each level the concentration
Name Level1 Level2 Level3
C-A 10 20 30
C-B 10 20 30
C-C 10 20 30

Next step is to specify in the sequence which calibration sample corresponds to which level, specified in the level column, just enter the level number there and the sample type will automatically be set to calibration sample

Calibration run 1: Level 1
Calibration run 2: Level 2
Calibration run 3: Level 3

When you now run the calibration samples, the results (area or height) for each calibration run will be stored at each corresponding level in the method.

OK?

[ydna6969]

Hi varossf,

Again thanks for your reply.

I understand what you are saying, this is a great help.
Although I still have afew questions.
If you have a dual active (standard and samples), to quantify do you require 2 standards (bracketing standards we usually use) to calibrate for each peak, as there are 2 different concentrations but only one level column in sequence

ie Level 1 standards = peak 1 (conc entered into level one column of method)
level 2 standards = peak 2 (concf entered inot level two column of method)

sequence example

Run type Level name

Calibration 1 std Peak 1
Calibration 2 std Peak 2
Unknown 0 n/a sample 1 (2 peaks)
Unknown 0 n/a sample 2 (2 peaks)
Unknown 0 n/a sample 3 (2 peaks)
Unknown 0 n/a sample 4 (2 peaks)
Unknown 0 n/a sample 5 (2 peaks)
Unknown 0 n/a sample 6 (2 peaks)
Calibration 1 std Peak 1
Calibration 2 std peak 2

Therefore 2 calibration curves for each bracket standard.
In the past I have used the same standard but this is the only way i think it is possible with this software, i could be wrong

Thanks

[varossf]

Ha, somehow I expected this question, but was not sure if I had to add the solution to my previous response.
In principle we speak about 4 levels here. 2 for Cal_1 and 2 for Cal_2. Smart readers understand in which direction we are going here.
Having your compounds separated into multiple physical samples does not mean that they have to be in the same 'level' in the method or sequence, that is only a logical reference.
So how do we solve this conundrum in EZChrom or the new OpenLAB EZChrom?

We need to specify 4 levels in the method
Compound name ; Level_1 ; Level_2 ; Level_3 ; Level_4
Std peak 1 ; 10; 20; ;
Std peak 2 ; ; ; 10; 20

So leave the levels where nothing is specified empty, do not fill out 0 here as 0 is considered a value in calibration.


Here is now your sequence below how it needs to be set up

Calibration 1 std Peak 1-----> Level_1 (ignoring peak 2 as that has no level_1 associated)
Calibration 2 std Peak 2-----> Level_3 (ignoring peak 1 as that has no level_3 associated)
Unknown 0 n/a sample 1 (2 peaks)
Unknown 0 n/a sample 2 (2 peaks)
Unknown 0 n/a sample 3 (2 peaks)
Unknown 0 n/a sample 4 (2 peaks)
Unknown 0 n/a sample 5 (2 peaks)
Unknown 0 n/a sample 6 (2 peaks)
Calibration 1 std Peak 1-----> Level_2 (ignoring peak 2 as that has no level_2 associated)
Calibration 2 std peak 2-----> Level_4 (ignoring peak 1 as that has no level_4 associated)

Now each compound will be calibrated accordingly and each unknown will be calculated correctly.
Bracketing is something you specify during sequence reprocess.

Does that help you?

[varossf]

We need to specify 4 levels in the method
Compound name ; Level_1 ; Level_2 ; Level_3 ; Level_4
Std peak 1 ; 10; 20; 0; 0
Std peak 2 ; 0; 0; 10; 20

please read

We need to specify 4 levels in the method
Compound name ; Level_1 ; Level_2 ; Level_3 ; Level_4
Std peak 1 ; 10; 20; ;
Std peak 2 ; ; ; 10; 20

please do not fill in 0 for those levels that should remain empty else the software will take 0 as a value. just keep them empty.

[varossf]

Hi ydna, and did you try it out already? Any more questions?

[ydna6969]

Hi varossf ,

Sorry for the late reply.
I have not tried this yet for dual active studt but does make sense, thanks for your help

What would you do if you had more known actives, lets say 4
Would this require 4 standards to bracket samples at the beginning and end ie

Std 1 (peak 1)
Std 2 (peak 2)
Std 3 (peak 3)
Std 4 (peak 4)
Sample 1
Sample 2
Sample 3
Sample 4
Std 5 (peak 1)
Std 6 (peak 2)
Std 7 (peak 3)
Std 8 (peak 4)

If this is the case run times would increase significantly ?

Also do you have any information on how compliant the software is ,in respect to CFR21 compliance ?

[varossf]

Hi,

if you split your compounds across multiple samples, like in your example 1 compound per sample, then indeed the total time for the sequence would increase as you would need 8 samples for 2 level calibration
If you combine the 4 compounds into 1 sample per level then you only have 2 samples, one sample for each level

Example given below for combined samples
Compound_Level1__Level2
CmpA_______10______20
CmpB_______12______18
CmpC_______8_______14
CmpD_______14______28

Sample_1 would be Level1
Sample_2 would be Level2

So you can mix and match as much as you like here.

Does that answer your questions?

Below case your example
Std 1 (peak 1)___________Level1
Std 2 (peak 2)___________Level2
Std 3 (peak 3)___________Level3
Std 4 (peak 4)___________Level4
Sample 1
Sample 2
Sample 3
Sample 4
Std 5 (peak 1)___________Level5
Std 6 (peak 2)___________Level6
Std 7 (peak 3)___________Level7
Std 8 (peak 4)___________Level8

[renibu]

hi,

i want to create an intelligent report:
where do I find the "STD ID #" and "STD Mult." in the menu selection?