First order derivative spectroscopy method involves the measurement of absorbance at zero crossing point of other drug. A new, rapid, precise, accurate and sensitive first derivative spectrophotometric method is proposed for the simultaneous estimation of Citicoline and Methylcobalamin in pharmaceutical dosage form. The drugs obeyed the Beer’s law in the concentration range of 10-20 ug/ml (r2 = 0.999) for Citicoline and 15-30 ug/ml (r2 = 0.999) for Methylcobalamin. In the proposed method, absorbance was measured at 267.86 nm (Zero Crossing Point of Metylcobalamin) for Citicoline and 270.89 nm (Zero Crossing Point of Citicoline) for Methylcobalamin. Accuracy of the method was determined by recovery studies and the average % recovery was found to be 99.97% and 99.92% for Citicoline and Methylcobalamin respectively. The LOD for Citicoline and Methylcobalamin were found to be 1.9041 μg/ml and 1.5617 μg/ml and LOQ were found to be 1.077 μg/ml and 1.07325 μg/ml respectively. Statistical analysis proves that the method is selective and sensitive for estimation of Citicoline and Methylcobalamin in pharmaceutical dosage form. The results were found to be within acceptance criteria according to ICH Q2 R1 guideline.
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