HPLC Method for Separation of 10 Water-Soluble Vitamins
High Performance Liquid Chromatography (HPLC) Method for separation of Vitamin B2 (Riboflavin), Nicotinic Acid/Niacin (3-pyridinecarboxylic acid), Nicotinamide, Vitamin B6 (Pyridoxine), Folic Acid, Cyanocobalamin, Thiamine diphosphate (Thiamine pyrophosphate), Vitamin B1 (Thiamine), Pantothenic Acid, Biotin
Riboflavin (B2), Nicotinic Acid (B3), Nicotinamide (B3), Pyridoxine (B6), Folic Acid (B9), Cyanocobalamin (B12), Thiamine Pyrophosphate (coenzyme of B1), Thiamine (B1), Pantothenic Acid (B5), Biotin (B7) are water soluble vitamins with a key function of energy metabolism. These coenzymes are responsible for converting food into usable energy.
Vitamin B2 (Riboflavin), Nicotinic Acid/Niacin (3-pyridinecarboxylic acid), Nicotinamide, Vitamin B6 (Pyridoxine), Folic Acid, Cyanocobalamin, Thiamine diphosphate (Thiamine pyrophosphate), Vitamin B1 (Thiamine), Pantothenic Acid, Biotin can be retained and analyzed using the Chromni™ stationary phase column. The analysis utilizes a gradient method with a simple mobile phase consisting of water, acetonitrile (MeCN). Detection is performed using UV.
Gradient Elution Program for HPLC Method
| Time (min) |
A – H2O (%) |
B – MeCN (%) |
C – H3PO4 1% in H2O (%)
|
Notes |
| 0 |
0 |
90 |
10 |
Starting Conditions (Hold) |
| 4 |
0 |
90 |
10 |
Gradient Start |
| 20 |
20 |
30 |
50 |
Gradient End |
| 20.01 |
0 |
90 |
10 |
Column Equilibration |
| 30 |
0 |
90 |
10 |
End of Run |
