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I'm injecting 20 microliters. I noticed that if I use 65% with 0.01M TBAB, the peaks look completely different, and I cannot tell if it's the ACN or the TBAB.How much you are injecting and what is the retention time? You might be seeing effect of large volume injection. Also, mixed-mode columns will allow you to avoid the use of IP reagent (it is attached to the surface of silica)