HPLC Separation of 2-HEP and 2-HPP in the Presence of Methylcobalamin and Amino Acids
https://sielc.com/Application-HPLC-Separation-of-2-HEP-and-2-HPP-in-the-Presence-of-Methylcobalamin-and-Amino-AcidsChromatogram
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Description
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.
Method Parameters
| 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 |
HPLC Column Used
Primesep SB, 4,6×150 mm, 5 µm, 100A