Chromatography Forum

Can anyone suggest a protocol for automatic precolumn derivatization of primary amines using the Agilent 1100 autosampler?

The one I've been using (below) is not giving good linearity and I dont know if it is due to the program I've developed (guessed!)

Thanks

John

Draw 0.5ul from buffer (borate pH 10)
Draw 0.5ul from sample
Mix 1.2ul 6 times in air
Draw 0.5ul from fluorescamine (in acetonitrile ~0.4mg/ml)
Mix 2ul 8 times in air
Draw 0ul from wash vial
Wait 2 min
Inject

Hi John

What is the concentration range ?

How good is the repeatability at any given concentration ?

Is the calibration curved, or just scattered ?

Is the deviation from linearity at high or low concentrations ?

My first impression is that the volumes are all small, and therefore vulnerable to large % variability.

Peter

Thanks for that Peter.

I've upped the volumes by a factor of 20 and will see how I get on.

It's a first shot in a troubleshooting exercise. I won't ahve time to do a full validation I'm afraid.

Cheers

John