Ion Pairing reagent/Phloxine B

Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.

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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?
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)
Vlad Orlovsky
HELIX Chromatography
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Vlad Orlovsky wrote:
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.
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