SummaryBacteriocins are antimicrobial peptides synthesised ribosomally by Gram‐positive or Gram‐negative bacteria to gain a competitive advantage. The majority of bacteriocins are derived from Gram‐positive bacteria, with lactic acid bacteria being the most common source. Because they are considered ‘natural’, there is currently significant development of bacteriocins for application as food preservative agents. As a preservative agent, bacteriocin activity is highly dependent on purity, down to the amino acid profile and sequence. Therefore, bacteriocin identification is important. Currently, MS is a cutting‐edge tool in bacteriocin identification. This method has high selectivity, sensitivity and resolution. To the best of our knowledge, systematic reviews focusing on the application of MS for bacteriocin identification are currently limited. In light of this, the objective of this study is to provide a comprehensive review and summary of MS technologies in bacteriocin research, with a particular focus on the discovery and characterisation of novel sources of bacteriocin. Additionally, studies related to the discovery of bacteriocins from various sources, their role as antimicrobial agents, and their synthesis are emphasised. Thus, this study presents a comprehensive analysis of the advantages, limitations, and future perspectives of the methods employed.