Quantitative Simultaneous Estimation of Water Soluble Vitamins, Riboflavin, Pyridoxine, Cyanocobalamin and Folic Acid in Neutraceutical Products by HPLC

Water soluble vitamins e.g. riboflavin (B2), pyridoxine (B6), cyanocobalamin (B12) and folic acid in neutraceutical product have been determined simultaneously by using a rapid, precise and time saving new high performance liquid chromatographic method and its validation. The method involves gradient elution of mobile phase through C18 discovery column (Supelco, Sigma-Aldrich) in a reverse phase chromatography with UV detection at 254 nm at ambient temperature. The ranges for quantification for B2, B6, B12 and folic acid were 0.13 mg g -1 (0.57-131 g g -1 ), 0.235 mg g -1 (3-235 g g -1 ), 7.94 x 10 -2 mg g -1 (8-80 g g -1 ) and 9.66 x10 -2 mg g -1 (10-97 g g -1 ), respectively. For the validation of the method, linearity, precision, accuracy and robustness have been performed. The repeatability was measured in terms of RSD value. The RSD for all vitamins was below 1%. Recovery of vitamins ranges from 98.6 to 100.5%.