Ion Pairing reagent/Phloxine B

[Andres760]

I'm testing for Phloxine B using 55% ACN with 0.03M TBAB at 548 nm. I'm getting peak splitting, could this be caused by too much ion pairing reagent, and what are the effects of using too much?

[Vlad Orlovsky]

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)

[Andres760]

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)

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.