Empower 2: Not Calibrating

[adurgan]

So, I am a new to this software but have been following the operations manual and learning about the program for the past week. I have collected different types of data and processed them without any problems.

Now, the senior researchers want me to begin using these other standards (ones that did not come with the instrument, at least to my knowledge). I have run them through using the proper methods and such (labeling them as standards, preping the column, etc.) and can integrate the chromatgrams just fine. The problem arises when I attempt to calibrate the peaks to form a calibration curve. The option is available but never produces any results.

I would really like some assistance on this matter, as I am running out of ideas on how to fix the situation. Thanks for all help.

Basic Running Parameters:
Eluent- 100% DI Water Flow (1ml/min)
Method- Condition Column, Purge Detector, Equilibriate, Inject Standard 1 x2, Inject Standard 2 x2, Inject Standard 3 x2.

[aceto_81]

A few things to check: Are you really sure you set the injections to standards?
Did you entered amounts for your components?
Did you entered the right names for your components (copy from processing method?)?

Gl

Ace

[adurgan]

I did enter the injections as standards, so that isn't the problem. However, the other questions you posed could be, can you elaborate further on what you mean?

Did you entered amounts for your components?
--Do you mean injection amounts, molecular weights, or something else? And where should I be inputing this information?

Did you entered the right names for your components (copy from processing method?)?
--What do you mean "the right names for your components" as in the names of the standards or the proper names of the methods for each of the injections I am trying to calibrate?

Thanks for your help and all other help.

Also, just to be specific, I am running a GPC instrument.

[aceto_81]

GPC or not, this has nothing to do with your type of LC, but with the used software.

Let clarify some stuff here:
I'm assuming you are in the pro interface of empower.
So, after you injected your samples, you go to browse project, you select your samples and then click on "Alter Samples" (if you cant find the icon in the bar, right click on your selected samples, and you will find it in the popup menu).
Now you can check wether or not your samples are set to standards or not.
Also, there is an icon in the bar that's called amounts.
Click on this, and normally you will see your entered amounts.
If not, you can click on "Copy from method set", or "copy from processing method", so that the component names from your processing method will apear in the amount table. Now put in the value you want to calibrate on for each component. Click ok, then save your samples. Exit of the alter sample screen. Goto review, open your processing method, click the calibrate button for your samples, and if everything goes fine, you will find a calibration curve in your calibration curve window!.

I suggest you will take some lessons from Waters, or at least browse thru the manuals of your Empower Software.

Good luck

Ace

[adurgan]

Thanks for all the help. Between my posting and your latest response, I found the answer online at http://www.forumsci.co.il/HPLC/gpc_steps.doc .

I still appreciate your help. The problem I was having is the operations manual I had apparently was missing some pages but I couldn't really tell.

I have calibrated most of the data already and am getting very good R^2 (0.9992 and better)

Thanks again!