HPLC Application for Simultaneous Separation of Amino Acids, Hydrophilic Acidic and Hydrophobic Neutral Compounds

Mixed-mode chromatography allows separating, in single run, compounds with vastly different properties. A method for separation of amino acids (cysteine, methylcysteine, cystine and dimethylcysteine) in the presence of carboxylic acid (benzoic) and hydrophobic neutral compounds was developed on Primesep 100 mixed-mode column. At lower pH ionization of carboxylic acids is suppressed. Amino acids are retained as basic compound based on reverse phase and cation exchange mechanisms. Carboxylic acids are retained on this column based on weak reverse phase mechanisms. Neutral compounds are retained by reverse phase mechanism as on any other column. Retention time of basic, zwitter-ionic and hydrophobic compound can be adjusted by manipulation of mobile phase composition. ELSD, UV or LC/MS detection can be used based on the properties of analytes and mobile phase selection.

Condition

Column Primesep 100, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer H2SO4
Flow Rate 1.0 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements, Amino acid
Analyzing Compounds Cysteine, Methylcysteine, Cystine, Dimethylcysteine,  Benzoic acid, Toluene,

Application Column

Primesep 100

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:
2,2-Dimethylcysteine
2-Methylcysteine
Benzoic Acid
Cystine
L-Cysteine
Toluene

Application 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.