December 23, 2024, 12:40:28 PM
Forum Rules: Read This Before Posting


Topic: HPLC calibration problem.  (Read 3468 times)

0 Members and 1 Guest are viewing this topic.

Offline mahrap

  • New Member
  • **
  • Posts: 6
  • Mole Snacks: +0/-2
HPLC calibration problem.
« on: June 14, 2013, 05:08:37 PM »
When I try calibrating my HPLC the areas of the curves of the different concentrations go down as the concentration goes up. What do you think is the cause of this problem? I know more concentrated solutions should give a bigger area but I get the exact opposite pattern.

Offline Arkcon

  • Retired Staff
  • Sr. Member
  • *
  • Posts: 7367
  • Mole Snacks: +533/-147
Re: HPLC calibration problem.
« Reply #1 on: June 15, 2013, 10:19:18 AM »
Well, try graphing the data by hand and look at what the curve shows.  Its possible that, instead of "going down", it may be leveling off, because the system is saturating at the higher end.  But that's just an example, we would need to see the data.  And its easier for us if you chart it.
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline pottesjaak on den hoak

  • New Member
  • **
  • Posts: 3
  • Mole Snacks: +0/-1
  • Gender: Male
Re: HPLC calibration problem.
« Reply #2 on: August 21, 2013, 04:06:39 PM »
perhaps there is a way around it,

I don't know what software your system runs on,

But whenever I encounter callibration problems i tend to cheat and outsmart the system,
if there is a mode in your software, you could manually integrate your peaks let it calculate the area for you,
and by writing down that very area, you can establish a series of different areas of different concentrations.
which you can fill down in you callibration table

that is only if the software supports manual integration ( and other ways of data analysis )


Offline Archer

  • Chemist
  • Sr. Member
  • *
  • Posts: 1001
  • Mole Snacks: +85/-20
  • Gender: Male
Re: HPLC calibration problem.
« Reply #3 on: August 22, 2013, 01:36:03 AM »

But whenever I encounter callibration problems i tend to cheat and outsmart the system


I take it your lab isn't accredited, if I discovered my staff doing this they would be given a verbal warning, second time would be written and third would be dismissal.

@Mahrap

This is the worst advice anyone can give you! If you have to 'fiddle' your calibration data how do you know whether or not to 'fiddle' your sample(s) which you are trying to quantify? Or will you just change it to suit the answer you were hoping for?

Let's see your data as Arkon suggests and see where we go from there.

Are you using an internal standard?

“ I love him. He's hops. He's barley. He's protein. He's a meal. ”

Denis Leary.

Sponsored Links