HPLC Separation of Two Vitamins: Different Polarity – Isocratic Methods
https://sielc.com/Application-HPLC-Separation-of-Two-Vitamins-Different-Polarity-Isocratic-MethodsChromatogram
Description
Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Peak order reversal is exhibited on these two columns with the same mobile phases due to their different polarity. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either formic or acetic acid (HOAc) with UV detection at 250 nm.
Method Parameters
| Mobile Phase | MeCN/H2O |
|---|---|
| Buffer | Formic Acid, Acetic Acid |
| Flow Rate | 0.5 ml/min |
| Detection | UV, 250 nm |
| Class of Compounds | Drug, Vitamin B₆, Hydrophobic, Ionizable |
| Analyzing Compounds | Vitamins C (ascorbic acid) and B2 (riboflavin) |
HPLC Column Used
Primesep B, Primesep B2 , 3.2×50 mm, 5 µm, 100A