Piperacillin belongs to a semisynthetic group of penicillins called ureidopenicillin that have been developed to have antipseudomonal broad spectrum activity to be used in the treatment of serious nosocomial infections, particularly when combined with a β-lactamase inhibitor as tazobactam. Piperacillin is used in treating susceptible bacterial infections, including skin and soft tissue, intra-abdominal, gynaecological, urinary tract and lower respiratory tract infections including acquired and nosocomial pneumonia. Moreover, it has veterinary applications either in treating bacterial infections or as growth-promoting agent in the livestock industry, causing the presence of piperacillin residues in different food samples obtained from animal origin in addition to its residues in various environmental samples. Because of the high clinical significance of the antipseudomonal penicillin, piperacillin, various methods have been reported for its determination in different matrices including pharmaceutical preparations, food, environmental and biological sample for studying pharmacokinetics or for therapeutic drug monitoring. Thus, this review summarizes different chemical and biological properties of piperacillin in addition to a comprehensive discussion of various analytical reports that have been published in the period 2000–2023 for piperacillin analysis. The discussed analytical reports encompasses six sections according to their different applications including population pharmacokinetic modelling, therapeutic drug monitoring, food analysis, livestock samples analysis, environmental analysis and pharmaceutical analysis with their related detailed tables. Moreover, current trends and future perspectives for piperacillin analysis have been suggested.