RESEARCH ARTICLE ISSN: 2348-8948 Vol: 2; Issue: 5 ANALYTICAL RP-HPLC METHOD FOR DEVELOPMENT AND VALIDATION OF CITICOLINE SODIUM AND METHYLCOBALAMIN IN COMBINED TABLET FORMULATION SHWEA D. SINGH * , FALGUN A. MEHTA 1 , DIMAL A. SHAH 2 , USMANGANI K. CHHALOTIYA 3 1 Department of Pharmaceutical Analysis, Indukaka Ipcowala College of Pharmacy, New Vallabh Vidayanagr, Anand, Gujarat, India. Date Received: 20 th Apr2014 Date of Accepted: 28 th Apr2014 Date Published: 12 th May2014 432 Abstract: Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method have been developed and validated for the estimation of Citicoline Sodium and Methylcobalaminin Bulk drug and Pharmaceutical dosage form. The developed method is rapid, accurate, precise, simple and economical than the previous methods. The separation was carried out usingPhenomenex Luna , C18, column having 250mm X 4.6mm, 5μm particle size, in isocratic mode, with mobile phase containing Acetonitrile: 0.02M KH 2 PO 4 [60: 40, v/v]. The flow rate is 1 ml / min and effluents are monitored at 554 nm. Chromatogram showed peak at a retention time of 3.8 min for Citicoline sodium and 2.3 min for Methylcobalamin . The method is validated for system suitability, linearity, precision, accuracy specificity, ruggedness, robustness, LOD and LOQ. Recovery of Citicoline Sodium and Methylcobalamin is found to be in the range of 99.76 - 101.79 % and 110.92% and 111.79%, respectively. The LOD and LOQ for estimation of Citicoline Sodium and Methylcobalamin are found to be 0.003 μg / ml, 0.0.01 μg / ml, and 0.01μg / ml, 0.05 μg / ml respectively. Proposed method can be successfully applied for the quantitative determination of Citicoline Sodiumand Methylcobalamin in Bulk drug and Pharmaceutical dosage form. Keywords: Citicoline (CITI), Methylcobalamin(MCA), RP-HPLC, Method Development, Validation. Introduction Citicoline is an intermediate in the generation of phosphatidyl choline from choline. It ischemically 5'-O [hydroxyl ({hydroxyl [2(trimethylammonio) ethoxy] phosphoryl}oxy) phosphoryl] cytidine. Citicoline is a white or offwhite amorphous, hygroscopic powder having molecular weight 488.3g/mol1. It helps toimprove focus and mental energy and may possibly be useful in the treatment of attention deficit disorder [1] . Methylcobalamin (MC) is a coenzyme of Vitamin B12 which is biologically active [2] . The importance of the combination of citicoline and methylcobalmine known to health care system to boost memory, production of brain energy, sustain cognitive function and motor skill and enhance focus and decision making thought processes. The major indication of the combination is diabetic neuropathy, diabetic gastropathy, memory loss, brain injury, anorexia, nerve rejuvenation, nerve regeneration and alertness [3] . Several method were cited in literature for determination of CITI and MCA individually or in combination with another drug by UV-Vis spectroscopy, HPLC, LC-MS and HPTLC but for combination only UV Spectrophotometric method was reported. Hence there is need to develop a simple, economic, precise and accurate high performance thin layer chromatographic method for Citicoline Sodium in the presence of methylcobalamin in tablet dosage form.