HPLC Analysis of Arginine and Related Impurities

Application description

Arginine is a semi-essential or conditionally essential amino acid which is zwitterionic in nature. It contains two carboxylic acid fragments and two primary amino groups. Meat and dairy products as well as grains, beans and nuts are a source for this amino acid. Arginine plays an important role in cell division, wound healing, various immune functions and the release of hormones. It is an important compound for the regulation of blood pressure. Like any other amino acid, it is not retained well in reversed-phase chromatography without the use of ion-pairing reagents which, in general, are not compatible with the mass spectrometry that is usually required in bio and chemical studies. Arginine and related impurities are retained by combination of weak reversed-phase and cation-exchange mechanisms. At a lower pH (below 3), the compound is more basic and retentive on this Coresep 100 mixed-mode reversed-phase cation-exchange column. The retention time for all compounds is adjusted by playing with acetonitrile, buffer pH and buffer concentration. Various buffers and acids can be used. The selection of the components of the mobile phase depends on the properties of analytes and the detection technique. The Coresep 100 column can be used for retention and separation of underivatized amino acids without ion-pairing reagent. In case of complex sample matrices like biofluids, additional sample preparation is required for developing a robust and reproducible method for analysis of underivatized amino acids

Conditions of Experiment
Column: Coresep 100
Separation Modes: reversed-phase, cation-exchange
Column Dimenstions: 4.6 x 150 mm, 2.7 um, 90A
Mobile Phase: ACN gradient from 2% to 20%, H2SO4 from 0.02% to 0.2% in 12 minutes
Detection: UV 210 nm
Sample: 0.3 mg/ml
Injection: 10 uL
Flow rate: 1.0 mL/min
Class of compounds: Amino acid, Aromatic acid, Supplement
Nature of compounds: Acidic, Hydrophilic, Polar, Zwitterionic
Compounds: Glutamic acid, Citrulline, Ornithine, Lysine, Arginine