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.
Separation of vitamins group B is achieved on Obelisc N column in HILIC mixed-mode. Vitamins of this group are different in polarity and ionic properties. Retention and separation is achieved by optimization of amount of ACN, buffer and buffer pH. Combination of UV and ELSD detection is used to monitor HPLC separation. B vitamins are water-soluble vitamins that play an important role in cell metabolism. Supplements containing all six are generally referred to as a vitamin B complex. Individual B vitamin supplements are referred to by the specific name of each vitamin. This method can be used to analyze individual B vitamins as well as vitamin B complex. Isolation of impurities as well as degradation products is possible by preparative chromatography.
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