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- Posts: 1
- Joined: Mon May 08, 2023 10:50 am
- Location: Belgium
I would like to ask your advise in achieving peak separation between two organic compounds: ribulose 1,5-biphosphate (RUBP) and 3-phosphoglyceric acid (3-PGA) in HPLC using ELSD as detector. In the literature, they achieved peak separation between the two compounds by using the following mobile phases:
Mobile A : 20 mM ammonium acetate and 20 mM ammonium hydroxide in water (pH 9.45)
Mobile B : acetonitrile (ACN):water (85:15, v/v)
Unfortunately, we only have the Shimadzu GIST NH2 column whose working pH range is limited within 2 - 7.5, hence I am very limited in adjusting the pH of my mobile phases. I have tried using just 50 mM ammonium acetate as mobile phase A without adjusting the pH. Unfortunately, I could not manage to achieve peak separation despite using gradient elution program.
I would like to still explore different ways to make it work with our current column and only buy one if it is really necessary (last option). However, I have limited experience with chromatographic separation, let alone for similar organic compounds.
Would anyone have some helpful advice for this?