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- Posts: 175
- Joined: Thu Jul 26, 2007 2:13 pm
The only potential issue that I thought might need to be addressed was that the UV wavelength I used was not the absolute maximum. The compound has two maxima, 232nm and 252nm. I chose 252nm because at the 232 wavelength the baseline drift when running the gradient is much greater and made the chromatography problematic. I decided to try KH2PO4 at pH 3.2 since it has a lower UV cutoff and this would eliminate the drift at the 232nm wavelength. (I tried to keep this as short as possible, but also wanted to get all the details).
Long story short, I made a 10mmol PPM buffer at 3.2 and completely lost retention for this main compound. How can this happen when this pH works at 3.2 in the formate buffer system?