Chromatography Forum

I have a problem compound that I can only get good chromatography for using this scenario:

20/80 H2O/ACN

Thermo Scientific Hypersil Gold, 2.1 x 50mm, 1.9µ

0.600mL/min flow rate

Retention time is 0.17 minutes

Total Run time is 3.0 minutes

I know this is very early, and I use gradient methods 99% of the time, but the peak shape is nice and the experiment passes. What would be an acceptable retention time in this case? Is any retention time acceptable when using isocratic?

RT 0.17min is at the void time of the column. What does the chromatoghraphy look like (tailing, fronting?) at lower %ACN? In the case of an unretained compound you might as well just dissolve your sample and measure the absorbance using a UV-vis spectrophotometer.