assess concentration using HPLC - (Nov/12/2008 )

Hi Everyone,

I need advice on HPLC (I've never used it!!)

As part of my PhD I need to assess the concentration of my test article (to ensure the concentration I said I'm going to use is the concentration that reaches the cells).

I have a pesticide (carbendazim) and it will be diluted in DMSO and added to cell culture media (DMEM). How can I set up a HPLC protocol to assess the concentration of pesticide in my treatment solution?

Thanks very much,

find the hplc procedure in the literature (which column or stationary phase, mobile phase, gradient or isocratic, detection method).

run a standard curve then your samples and compare to the standard curve.

I seem to recall that pesticides are often used by HPLC column manufacturers to show the separatory ability of their columns (at least I ran across a few when investigating reverse phase columns from Varian and GE Healthcare/Amersham awhile ago).

As far as determining the concentration, most HPLC software can integrate the area under a peak, which I think will give you concentration assuming you have a clean peak with no shoulders.