|Molecular Weight||169.180 g/mol|
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.
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 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