building a calibration curve

[kolibri]

Can somebody tell me how to have the calibration curve on my HPLC results. I have run an HPLC with calibration and changed in analysis method the result option in % normalization. But in the end I still dont have the calibration plot... Thanks!

[Ron]

There isn't much information to go on here, but if you have specified area normalization there is no calibaration for this in most data systems, it just gives area percent of the components detected. Please provide more specifics of you data system and application.

[kolibri]

Thank you for your reply. Well I have specified according to the manual the %normalization and named the peaks. But actually there is no other information on how to get to the calibration results, only how to open the already existing plot. So I dont know how to get to the results where it gives me the calibration curve from the run...

[JGK]

Thank you for your reply. Well I have specified according to the manual the %normalization and named the peaks. But actually there is no other information on how to get to the calibration results, only how to open the already existing plot. So I dont know how to get to the results where it gives me the calibration curve from the run...

We still need more information !

What software system are you using? If we know this we can provide the help you need much more easily.

In most cases, %normalisation will only tell you what percentage area each peak has based on the total peak area of all integrated peaks in the chromatogram it will not perform a calibration for you.

Is there an option for "External Standards"? This type of option will allow you to enter the identity (vial No., concentration etc) of your calibration STDs in order to construct a calibration line for sample measurement.

[kolibri]

I am using Gilson HPLC with Unipoint V 1.71 software. If you could tell me a step by step process how to make a calibration curve it would be very helpfull. Thank you again very much..

[JGK]

I am using Gilson HPLC with Unipoint V 1.71 software. If you could tell me a step by step process how to make a calibration curve it would be very helpfull. Thank you again very much..

Sorry, I have no experience of this software system.

Do you have any User Manuals?

Have you tried using the on-line Help in the program?

Also, was there any form of Tutorial disk supplied with the software?

Any of these sources may help you to get started. Failing that, try contacting your Gilson sales rep ot their technical support.

[kolibri]

thank you very much for your reply.
I have checked in the manual there is no information about this. I am not the first one to use it so I'm afraid that there are no CD-s available with the manual. But there is also a possibility that the program is too old for a CD:) I will try to find an online information. Thank you again.

[DR]

Open up Excel
Put your standard concentrations in a column, put the corresponding areas in an adjacent column
Click and drag over both and make an X-Y scatter plot
select series
add trend line
format trend line to show equation

Once you figure out how to get the CDS to match this, you're home-free.