The components of vitamin B₆ family resemble each other in structure and are difficult to separate. Usually, the method using ion pair reagent is used. However, under strong acidic conditions such as at pH 2.0, separation is possible by reversed phase mode using the slight difference of hydrophilicity. Three components of vitamin B₆ family were separated with the Asahipak ODP-50 6D column, which can be used even at pH2.0.


Sample : 10 μL
1. Pyridoxamine 2HCl 0.1 mg/mL
2. Pyridoxal HCl 0.2 mg/mL
3. Pyridoxine HCl (Vitamin B₆) 0.12 mg/mL

Column       : Shodex Asahipak ODP-50 6D (6.0 mm I.D. x 150 mm)
Eluent       : 50 mM Sodium Phosphate buffer (pH2.0)
Flow rate    : 1.0 mL/min
Detector     : UV (254 nm)
Column temp. : 30 °C