A sample rich in anthocyanins has been passed through UHPLC-DAD, using a gradient with acidified water (formic acid 0.1%) and methanol and a flow rate of 0.5 mL/min. However, the chromatogram is not clear and any peak appears. What am I doing wrong?
It might be because the sample injection volume is less. Increase the injection volume of your sample and the gradient should be with high water and low methanol percentage.
Nitisha, thank you for your reply. I will try to modify the injection volume and the gradient.
Dear Maribel,
The extraction protocol is also an important step in order to observe clear peaks on the chromatogram for the anthocyanins. I may suggest the following paper for the procedure: 10.1016/j.ultsonch.2018.07.007
In this study, I obtained separate peaks for anthocyanins.
Best.
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