HPLC UV Method for Separation of NAD and NADH on PEI Column
High Performance Liquid Chromatography (HPLC) Method for Analysis of SNAD and NADH
Nicotinamide adenine dinucleotide (NAD), is a coenzyme found in every single living cell. NAD can exist in two forms: NAD+ and NADH. The conversion of NAD from its oxidized form (NAD+) to its reduced form (NADH), and back, provides the cell with a mechanism for accepting and donating electrons.
NAD and NADH can be retained, separated and UV detected at 264 nm using the PEI column with a simple mobile phase of acetonitrile (ACN) and water with sulfuric acid buffer and detected by UV.
Methodenparameter
| Column | PEI, 4.6 x 50 mm, 5 µm, 100 Å, |
|---|---|
| Mobile Phase | MeCN/H2O – 60/40% |
| Buffer | H2SO4 – 0.2% |
| Flow Rate | 1.0 mL/min |
| Detection | UV 264 nm |