Previously, we reported the synthesis of stable gold nanorods with higher aspect ratio (approximately 15-20) and more enhanced uniformity by a modified seed mediated growth approach using a binary surfactant system consisting of CTAB and Pluronic F-127. For the in vivo application of gold nanorods, the removal of CTABs that are strongly bound on prepared Au nanorods is necessary due to their cytotoxicity. Use of heat or acid at various conditions was performed to achieve the complete removal of CTAB from the synthesized Au nanorods while maintaining their stability. Here, we report the appropriate conditions for both treatments that can remove CTAB efficiently without hampering the stability of Au nanorods. After the removal of CTAB, Pluronic F-127 was added additionally for the stabilization and further potential functionalization of Au nanorods to make useful for various applications. VIS-NIR absorption spectroscopy, SEM, TEM, and FTIR were used for characterization.