Separation of arginine and glutamic acid without derivatization using HPLC-UV

 1- Mobile phase preparation :

- Eluent (A) : potassium dihydrogen phosphate, 25 mM, pH 2.85

- Eluent (B) : Acetonitrile

- Eluent (A) : Eluent (B)

            25   :   75

2- Column : Acclaim Mixed-Mode HILIC-1

( 25 cm x 4.6 mm x 5 µm )

3- Column oven temp. : 30 °C

4- Detector : UV 200nm

5- Injection volume : 20 µL

6 - Chromatogram :

- Peak 1 : Glutamic acid

- Peak 2 : Arginine

- Peak 3 : Impurity