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.
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.
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.
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
High Performance Liquid Chromatography (HPLC) Method for Analysis of Pyridoxine Hydrochloride (Vitamin B6) and Benfotiamine
Pyridoxine Hydrochloride (also known as Vitamin B6), is a naturally occurring vitamin that can be found in a wide variety of common foods, such as meat, grains, and avocados. Benfotiamine is a synthetic form of Vitamin B1 that has shown early signs of treating nerve damage, Alzheimer’s, alcohol dependence. They can both be found in Milgamma 100 tablets, a medicine prescribed to treat Vitamin B1 and B6 deficiency.
These two vitamins can be detected in the low UV regime. Using a Primesep S normal-phase column and a mobile phase consisting of water and acetonitrile (MeCN) with a phosphoric acid (H3PO4) buffer, Pyridoxine Hydrochloride and Benfotiamine can be retained, separated, and analyzed. This analysis method can be UV detected at 280 nm.