Chromatography Forum

Hi all.
Iam trying to optimize 1,2,4-Triazole (T) on Agilent 6460 MS/MS detector.
Solvents/flow are A 90% H2O 0.01% Formic acid, 5mM Amonium formate
B 10% ACN 0.01% Formic acid
Column: Short C18 (20mm) 0.3 ml/min
When i inject 10ug/ml 1,2,4-Triazole in mobile phase i get nice peak
When i inject 10ug/ml 1,2,4-Triazole in ACN/H2O 1:1 i get negative peak no signal for 70m/z
Any idea why ?
Thanks for help

I would think that your problem is too much organic in the injection solution. If you have too high of a concentration of organic you will not be able to retain compounds on the column. If you need to inject this matrix, then you need to find a way to dilute the sample down before it hits the column. When I encounter this problem on a Waters system, my trick is to inject into a much larger sample loop than what would be required for the injection volume. This way the sample gets diluted with the mobile phase before it hits the column.