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Boldenone (BOL) has been frequently detected in doping and food safety over the past few decades. Researchers have studied BOL metabolism across various species, reporting significant differences even within the same species due to variations in experimental designs and analytical methods. Additionally, detection methods face challenges such as matrix interferences and the presence of endogenous structural analogs at low concentrations. This study aims to compile and analyze the development of chromatographic techniques for detecting BOL and its metabolites in biological matrices. An integrative review of literature from May 2000 to September 2024 was conducted using databases like PubMed, ScienceDirect, Elsevier, Springer, Scopus, Wiley, and Taylor & Francis. The MeSH terms ‘boldenone’ AND ‘detection,’ restricted to titles or abstracts, yielded 167 records, with 79 meeting the inclusion criteria. Hyphenated techniques (e.g., LC/MS/MS and GC/C/IRMS) were predominantly used and generally successful in identifying BOL, its precursors, and metabolites, particularly in characterizing their endogenous origin or differentiating isomers. Urine was the most commonly observed matrix, and solid-phase extraction (SPE) was the predominant extraction technique. Future research should aim to improve extraction and detection methods to address current discrepancies in controlling BOL use, as its pharmacological properties have led to negative repercussions in sports and concerns about food safety.