HPLC Separation of 2-HEP and 2-HPP in the Presence of Methylcobalamin and Amino Acids

chr_292.gif Methylcobalamin, S-Adenosyl-L-methionine, 2-hydroxyethyl phosphonic acid, and 2-hydroxy propyl phosphonic acid were separated in reversed-phase anion-exchange mixed-mode on a Primesep SB HPLC column. Compounds are monitored by UV and ELSD. Method is compatible with LC/MS. Selectivity of separation is controlled by varying the amount of ACn buffer pH and buffer concentration.


Column Primesep SB, 4,6x150 mm, 5 µm, 100A
Mobile Phase MeCN - 20%
Buffer AmFm
Flow Rate 1.0 ml/min
Detection ELSD 35C,  UV 250nm


Class of Compounds  Acid,  Hydrophilic, Ionizable
Analyzing Compounds S- Adenosyl- L- methionine, Methylcobalamin, 2-hydroxyethylphosphonic acid, 2-hydroxypropylphosphonic acid

Application Analytes:
2-hydroxyethyl phosphonic acid
2-hydroxypropyl phosphonic acid