Hi everyone.
I'm having some trouble with the peak shape on VancoShell column. The mobile phases used are 5,5 mM ammonium acetate (pH 4,7) and methanol, in the ratio 20:80. Column temperature is 10°C, and flow rate is 0,17 mL/min. I'm trying to develop LC-MS method for chiral separation of venlafaxin. The instrument is Agiletn 1290 UHPC coupled to Agilent QTOF 6550. In the beginning, the peak shape was OK, but at some point, the peaks started to have shoulders. I tried washing the column with pure methanol, as well as tried to regenerate the column as per instructions. It helped a little, but the peak shape is still not that great.
I'm not sure what happened with the column, since the only thing injected onto it was a standard solution. Does anyone have any advice on how to improve the peak shape? Also, I'm wondering can I reverse flush the column since there is nothing in the manual about that.
Noel W Davies I'm thinking the issue could be because of the void volume or maybe clogged frit. But I don't know can this column be reverse flushed since there is nothing about it in the manual.
Please provide the EXACT column dimensions for your chiral antibiotic HPLC column.
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