HPLC Separation of Lysine and Arginine from Other Amino Acids


Application Notes: Amino acids are polar ionic compounds which are not retained on reversed-phase column without ion-pairing reagent. In our application, lysine and arginine can be separated from other amino acids. Amino acids with a pH between 3 and 5 and with one basic and one acidic group become very polar. Therefore these amino acids don’t have strong ion-exchange interaction with Primesep C stationary phase. Amino acids with two amino groups still carry positive net charge and can interact with stationary phase by cations-exchange mechanism. pH variation of the mobile phase can be an effective tool to adjust selectivity of separation for zwitter-ionic, basic and acidic compounds. This method can be used for separation of mono-charged compounds from compounds having an extra charge.

Application Columns: Primesep C
Application compounds: Aspartic acid, Glutamic acid, Aspargine, Glycine, Proline, Alanine, Phenylalanine, Tyrosine, Leucine, Lysine, Arginine
Detection technique: UV, LC/MS, ELSD/CAD


Column Primesep C, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN – 15%
Buffer AmAc pH 5.0- 15 mM
Flow Rate 1.0 ml/min
Detection ELSD



Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds Aspartic acid, Glutamic acid, Aspargine, Glycine, Proline, Alanine, Phenylalanine, Tyrosine, Leucine, Lysine, Arginine


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.

Primesep C

The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.

Select options
Application Analytes:
Aspartic Acid
Glutamic Acid

Application Detection:
UV Detection
ELSD/MS Detection