Chromatography Forum

I have a colleague who is trying to develop an HPLC method for cyclopentolate hydrochloride with possibility to detect 0.1% degradation products. Does anyone have any suggestions for obtaining correct peak shape and sensitivity? He has been trying with octane sulfonate + triethylamine pH 3/ acetonitrile on a Uptispher column.
A second question concerning sensitivity; we are now systematically asked to detect 0.1% degradation products and this often means injecting very concentrated solutions giving rise to overloading, peak deformation and other chromatographic phenomena. Does anyone have any suggestions for increasing sensitivity without overloading?

Some overloading phenomena are related to the injection solvent and can be solved by changing this. Some peak distortion phenomena can be solved by using modern tailing-free columns. Some peak distortion phenomena can be reduced at least 10 fold by changing the pH, i.e changing your analyte to a non-ionic form.

Beyond this, only better detectors can help.