This document describes the separation method of Vitamin B6 complex on Pursuit Diphenyl column. The mobile phase consists of 5 mM ammonium formate and CH3CN in a ratio of 95:5. The flow rate is 0.5 mL/min and detection is performed at 254 nm. This method is suitable for the separation and detection of pyridoxamine, pyridoxal, and pyridoxine.
This document provides an application note for the Agilent 2236 Liquid Chromatography system. It details the specific method for separating a Vitamin B6 complex, which includes Pyridoxamine, Pyridoxal, and Pyridoxine.
The recommended method utilizes a Pursuit Diphenyl column with dimensions of 250 x 4.6mm and a 5-micron particle size. The mobile phase consists of 5mM Ammonium formate at pH 2.65 mixed with Acetonitrile (MeCN) in a 95:5 ratio. The analysis is performed at a flow rate of 0.5 mL/min and a temperature of 40°C, with detection at 254 nm.
| Column | Pursuit Diphenyl, 250 x 4.6mm, 5u |
|---|---|
| Mobile Phase | 5mM Ammonium formate, pH 2.65: MeCN - 95:5 |
| Flow Rate | 0.5 mL/min |
| Temperature | 40°C |
| Detection | 254 nm |
The complex includes Pyridoxamine, Pyridoxal, and Pyridoxine (or Pyridoxol or Vitamin B6).
The recommended flow rate is 0.5 mL/min.