HPLC UV Method for Separation of NAD and NADH on PEI Column

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Chromatogram

Description

· 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.

Method Parameters

Mobile PhaseMeCN/H2O – 60/40%
BufferH2SO4 – 0.2%
Flow Rate1.0 ml/m
DetectionUV 264 nm
Class of CompoundsDrug
Analyzing CompoundsNicotinamide Adenine Dinucleotide (NAD),Nicotinamide Adenine Dinucleotide (reduced) (NADH)

HPLC Column Used

PEI, 4.6 x 50 mm, 5 µm, 100 A,