HPLC Separation of Polyamines on Primesep 500 Column


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Polyamines are both naturally occurring and synthetic. The naturally occurring polyamines have a variety of functions including ion channel modulation and biosynthesis of cells. The synthetic polyamines are used in motor oil and epoxy resins. Because of their strong ionic characteristics, they are not retained on reverse-phase columns and strongly retained on anion-exchange columns. In order to separate polyamines with varying numbers of ionic groups, we used a Primesep 500 HPLC mixed-mode column with embedded carboxylic groups of pKa around 5. By lowering the pH of the mobile phase, we can suppress the carboxylic groups on the column making it less retentive to polyamines. We separate the double and triple charged polyamines at one pH, then do a step gradient to lower pH to reduce the retention time and separate higher charged polyamines. The detection can be achieved with CAD or MS.

 

Condition

Column Primesep 500, 4.6×150 mm, 5 µm, 100A
Mobile Phase H2O
Buffer AmFm pH 3.0 –  20 mM,  AmFm pH 3.5 –  20 mM
Flow Rate 1.0 ml/min
Detection CAD

Description

Class of Compounds Hydrophilic, Amines
Analyzing Compounds Ethylenediamine, N-(3-Aminopropyl)-1,3-propanediamine,  N,N’- Bis(3-aminopropyl)-1,3-propanediamine, Tetraethylenepentamine, Pentaethylenehexamine

Application Column

Primesep 500

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:
Ethylenediamine
N,N’-Bis(3-aminopropyl)-1,3-propanediamine
N-(3-Aminopropyl)-1,3-propanediamine (Norspermidine)
Pentaethylenehexamine
Polyamines
Tetraethylenepentamine
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.