HPLC Base Line Issue
In my HPLC Agelent 1260 infinity model, base line going to upward direction and my buffer is Ammonium Bicarbonate with pH - 8 and mixed with MeOH. Run time is 25 minutes.
Method:
0 minute: Ammonium Bicarbonate - 85% & MeOH- 15%
10 minutes: Ammonium Bicarbonate - 10% & MeOH- 90%
20 minutes : Ammonium Bicarbonate - 20% & MeOH- 80%
25 minutes: Ammonium Bicarbonate - 85% & MeOH- 15%
My diluent is DMSO
Please, give me the solution
1 Answers
Your Answer
Log in to answer this question.
3
Run a blank gradient (no injection): If the baseline still climbs, it’s a solvent/background issue, not sample/diluent.
Degas thoroughly: Use the 1260’s online degasser + pre-sonicate. Helium sparging if available. Make fresh ammonium bicarbonate (≤10–20 mM), filter 0.2 µm, and keep bottles capped.
Thermostat the column & flow cell: Set column oven (e.g., 30 °C). Keep the detector cell and solvents at stable room temp; avoid drafts/heat. wevlenght 360 to380 nm
Degas thoroughly: Use the 1260’s online degasser + pre-sonicate. Helium sparging if available. Make fresh ammonium bicarbonate (≤10–20 mM), filter 0.2 µm, and keep bottles capped.
Thermostat the column & flow cell: Set column oven (e.g., 30 °C). Keep the detector cell and solvents at stable room temp; avoid drafts/heat. wevlenght 360 to380 nm
