HPLC Method for Analysis of Water-Soluble Vitamins (B1, B2, B3, B6) on Primesep 100 Column

High Performance Liquid Chromatography (HPLC) Method for Analysis of Water-Soluble Vitamins (B1, B2, B3, B6) on Primesep 100 by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode SIELC Technologies

HPLC Method for Analysis of Vitamin B1 (Thiamine), Vitamin B2 (Riboflavin), Nicotinamide, Vitamin B6 (Pyridoxine), Pyridoxamine, Pyridoxal Phosphate on Primesep 100 Column


 High Performance Liquid Chromatography (HPLC) Method for Analysis of Vitamin B1 (Thiamine), Vitamin B2 (Riboflavin), Nicotinamide, Vitamin B6 (Pyridoxine), Pyridoxamine, Pyridoxal Phosphate

Water-Soluble Vitamins (B1, B2, B3, B6) can be retained, separated and analyzed using a Primesep 100 mixed-mode stationary phase column. The analysis employs an gradient method with a simple mobile phase comprising water, acetonitrile (MeCN), and sulfuric acid as a buffer. This method allows for detection using UV 300 nm.

You can find detailed UV spectra of Vitamin B1 (Thiamine), Vitamin B2 (Riboflavin), Nicotinamide, Vitamin B6 (Pyridoxine), Pyridoxamine, Pyridoxal Phosphate and information about its various lambda maxima by visiting the following links: Vitamins (B1, B2, B3, B6)

ColumnPrimesep 100, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseMeCN – 10%
BufferGradient H2SO4 -0.1-0.5% in 10 min
Flow Rate1.0 ml/min
DetectionUV 270 nm

Class of Compounds
Water-Soluble Vitamins (B1, B2, B3, B6)
Analyzing CompoundsVitamin B1 (Thiamine), Vitamin B2 (Riboflavin), Nicotinamide, Vitamin B6 (Pyridoxine), Pyridoxamine, Pyridoxal Phosphate

Application Column

Primesep 100

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
Nicotinamide
Pyridoxal Phosphate
Pyridoxamine
Vitamin B1 (Thiamine)
Vitamin B2 (Riboflavin)
Vitamin B6 (Pyridoxine)
Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.