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

Water soluble vitamins e.g. riboflavin (B 2 ), pyridoxine (B 6 ), cyanocobalamin (B 12 ) 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 C 18 discovery column (Supelco, Sigma-Aldrich) in a reverse phase chromatography with UV detection at 254 nm at ambient temperature. The ranges for quantification for B 2 , B 6 , B 12 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%.