The present manuscript describes a simple, sensitive, rapid, accurate, precise, and economical Simultaneous equation method and first-order derivative spectrophotometry method for the simultaneous determination of Chlorzoxazone and Tramadol in laboratory mixture. The absorbance values at 243.3 nm and 271 nm for the simultaneous equation method and 236.6nm and 213.3nm for the first derivative spectrum were used for the estimation of Chlorzoxazone and Tramadol. This method obeyed beer’s law in the concentration range of 2-10 μg/ml for Chlorzoxazone and 10-100 μg/ml for Tramadol. The solvents used for UV-Spectrophotometric methods was 0.1 N NaOH. The % RSD of accuracy was found to be 0.2086 for Chlorzoxazone and 0.4717 for Tramadol. The method was successfully applied to the laboratory prepared mixture because no interference from the mixture excipients was found. The suitability of this method for the quantitative determination of Chlorzoxazone and Tramadol was proved by validation. The results of analysis have been validated statistically and by recovery studies.