Preparation of fish tissue samples prior to LC-MS analysis can be a complex and time consuming process. Automation of this sample preparation process was investigated as an alternative to manually performing this workflow to make the process easier and faster. An automated workflow was utilized to extract, clean up and analyze fish tissues. An automated solvent extraction system was used for the extraction of samples following the guidance of EPA method 1633. Sample clean-up was then fully automated using an automated SPE system and SPE cartridges containing both WAX and GCB sorbents. Targeted LC-MS/MS analysis of 40 PFAS compounds was performed using a highly sensitive mass spectrometer. The automated workflow provided equivalent quality data when compared to utilization of the workflow manually. Additionally, certified reference materials were processed and analyzed using this workflow with all results falling within the acceptance criteria of each reference material. The automated solvent extraction system was able to reduce sample extraction times to approximately 6 minutes each. Manual extraction steps for soil samples require 95 minutes or more and over 16 hours for tissue samples. This significantly reduces the time spent on extraction of samples. Additionally, the automated SPE system processes a batch of samples (each ~50 mL) in approximately one hour. Finally, LC-MS/MS sample analysis was performed using an 11 minute method. The reduction of time spent prepping samples allows the full extraction to analysis workflow to be possible in a typical 8 hour shift while maintaining precision, accuracy and confidence in final results.