In view of the environmental repercussions of industrial livestock farming, alternative meat (AM) products have been gaining traction in the global market. However, progress remains slow due to a mismatch of the umami taste profile with conventional meat products. Thus, the development of an analysis platform capable of assessing the umami of meat analogues is critical to consumer palatability and acceptance. Therefore, in this study, an extraction and analysis method were developed to quantify the umami free amino acid and free 5’-nucleotide content in conventional and alternative meat samples. An extraction method using water was developed to mimic the release of umami compounds in the oral environment, while hydrochloric and perchloric acid were used in separate methods to obtain the full profile of the compounds. Separation, identification and quantification of the umami taste compounds using reversed-phase HPLC was achieved and successfully applied in combination with analysis via electronic-tongue to determine the Equivalent Umami Concentration (EUC) of beef and three AM samples. The taste profiles of all the four samples were found to be significantly different and varied significantly across the different extraction procedures, with the oral simulation extraction method achieving 75% extraction efficiency of the full umami compound profile. The umami rankings determined by the e-tongue agreed with that of chemosensory tests: mycoprotein < beef < plant-based meat alternatives (PBMA). The numerical EUCs determined by e-tongue and HPLC were also found to be similar for the mycoprotein and PBMA samples.