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

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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.

Condition
Column Primesep SB, 4,6×150 mm, 5 µm, 100A
Mobile Phase MeCN – 20%
Buffer AmFm
Flow Rate 1.0 ml/min
Detection ELSD 35C,  UV 250nm

 

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

Application Column

Primesep SB

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
2-hydroxyethyl phosphonic acid
2-hydroxypropyl phosphonic acid
Methylcobalamin
S-Adenosyl-L-methionine
Application Detection:
ELSD Detection
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.