Amitriptyline Hydrochloride (AMI) is a tricyclic antidepressant drug. The aim of this work is to develop a simple, accurate, reproducible and cost effective spectrophotometric method for determination of Amitriptyline Hydrochloride in a pharmaceutical dosage. The absorption maximum of this drug was at 239.0 nm and method has followed linearity in the concentration range 4-24 μg/mL. The generated regression equation was ABS = 0.043 C + 0.035 with R2 value as 0.9995. The limit of detection (LOD) and limit of quantification (LOQ) were 0.1094μg/ml & 0.3315μg/ml, respectively. Percent relative standard deviation values for the intra-day and inter-day precision were 0.215 and 0.452, respectively, indicated good precision. Results of the recovery studies showed good accuracy of the method. Statistical analysis of results suggests that the developed method is suitable for routine estimation of Amitriptyline Hydrochloride in pharmaceutical formulations.