HPLC Separation of 2-HEP and 2-HPP in the Presence of Methylcobalamin and Amino Acids using Zodiac HST-SB Column

Methylcobalamin, S-adenosyl-L-methionine, 2-hydroxyethyl phosphonic acid, and 2-hydroxypropyl phosphonic acid were separated using the Zodiac HST SB mixed-mode HPLC column, which combines reversed-phase and anion-exchange mechanisms. Detection was achieved using both UV and ELSD, and the method is fully compatible with LC/MS. Separation selectivity can be finely tuned by adjusting the acetonitrile (ACN) content, buffer pH, and buffer concentration.