CAS Number 65-23-6
Molecular Formula C8H11NO3
Molecular Weight 169.180 g/mol
LogP -0.770
  • Pyridoxine
  • 4,5-Bis(hydroxymethyl)-2-methylpyridin-3-ol
  • 3,4-Pyridinedimethanol, 5-hydroxy-6-methyl-
  • 65-23-6
  • 3,4-Pyridinedimethanol, 5-hydroxy-6-methyl-
  • 2-Methyl-3-hydroxy-4,5-di(hydroxymethyl)pyridine
  • 2-Methyl-4,5-bis(hydroxymethyl)-3-hydroxypyridine
  • 5-Hydroxy-6-methyl-3,4-pyridinedimethanol
  • Adermin
  • Bezatin
  • piridoxinio
  • Pirivitol
  • Pyridoxin
  • Pyridoxol
  • EINECS 200-603-0
  • 2-Methyl-3-hydroxy-4,5-bis(hydroxymethyl)pyridine
  • 2-Picoline-4,5-dimethanol, 3-hydroxy-
  • 2-Methyl-3-hydroxy-4,5-dihydroxymethyl-pyridin
  • 2-methyl-3-hydroxy-4,5-dihydroxymethylpyridine
  • 2-methyl-4,5-dimethylol-pyridin-3-ol
  • 3-Hydroxy-2-Picoline-4,5-dimethanol
  • 3-hydroxy-4,5-dimethylol-alpha-picoline
  • 4,5-bis(hydroxymethyl)-2-methyl-pyridin-3-ol
  • 5-hydroxy-6-methyl-3,4-pyridinedimethanol
  • Adermine
  • Gravidox
  • Hydoxin
  • Piridossina
  • Piridoxina
  • Pyridoxinum
  • Pyridoxolum
  • vitamin B6


HPLC Separation of Vitamin B
Primesep C separates B vitamins with baseline resolution by a combination of cation exchange, complex formation, and hydrophobic interactions. Vitamin B1 (thiamine), vitamin B2 (riboflavin), and vitamin B6 (pyridoxine) are separated with a mobile phase of water, acetonitrile (MeCN, ACN) and triethylamine (TEA) phosphate with UV detection at 280 nm.

Application Analytes:

Riboflavin (Vitamin B2)
Thiamine (Vitamin B1)
Vitamin B1 (Thiamine)
Vitamin B2 (Riboflavin)

HPLC Analysis of Active Drug in a Formulation

HPLC method for separation of active ingredients of drug/supplemental composition was developed on an Obelisc R trimodal HPLC column. Compounds are retained by combination of reversed-phase, cation-exchange and anion-exchange mechanisms. Compounds are well separated, and method can be used for quantitation of pyridoxine, ascorbic acid, niacinamide, pantothenic acid, caffeine and riboflavin in a mixture or as separate compounds in various complex mixtures. Various detection techniques can be applied for quantitation (ELSD, UV, LC/MS, Corona). This HPLC method can be adopted as general approach for analysis of active drug components in various formulations.

Application Analytes:

Ascorbic Acid
Pantothenic Acid
Riboflavin (Vitamin B2)

HPLC Separation of Vitamin C, Vitamin Group B, and Related Impurities

Vitamin C (ascorbic acid) and Vitamins Group B are separated on Obelisc N mixed-mode column. Method can be used in quantitation and determination of polar vitamins in various formulations and dietary supplements. HPLC method can be based on UV, Evaporative Light Scattering Detection (ESLD), RI or MS detection. Effect of sample matrix can be eliminated by changing mobile phase conditions. Buffer concentration, buffer pH and amount of ACN will affect every vitamin differently due to difference in polar and ionic properties.

Application Analytes:

Ascorbic Acid
Barbituric Acid
Isonicotinic Acid
Nicotinic Acid
Vitamin B2 (Riboflavin)
Vitamin B6 (Pyridoxine)

USP Methods for the Analysis of Pyridoxine for the Legacy L1 Column
  Application Notes: Pyridoxine is part of the vitamin B complex group. Pyridoxine is important in the body’s daily function as it regulates many enzymatic reactions. The USP HPLC method for the separation of pyridoxine was developed on Legacy L1 column according to the US Pharmacopeia methodology. L1 classification is assigned to reversed-phase HPLC column containing C18 ligand. Support for the material is spherical silica gel with particles size 3-10 um and pore size of 100-120A. Resolution between critical pairs corresponds to rules and specifications of UPS.   Application Columns: Legacy L1 C18 HPLC column Application compounds: Pyridoxine Mobile phase: AcOH/MeOH/H2O/Hexanesulfonate (2/47/153/1.2) Detection technique: UV Reference: USP35- NF30


Column Legacy L1, 4.6x250 mm, 5 µm, 100A
Mobile Phase AcOH/MeOH/H2O/Hexanesulfonate 2/47/153/1.2
Buffer Hexanesulfonate
Flow Rate 1.0 ml/min
Detection UV, 280 nm


Class of Compounds Drug, Vitamin B₆, Hydrophobic, Ionizable
Analyzing Compounds Pyridoxine

Application Analytes:

Vitamin B6 (Pyridoxine)