Abstract: As the predominant toxic constituent within the Aconitum genus, Aconitum alkaloids (ATs) exhibit both significant pharmaceutical value and substantial toxicity, have been widely used in traditional Chinese medicine and the realm of contemporary clinical medicine. However, owing to their high toxicity, inappropriate employment of ATs in pharmaceuticals, edibles, and the environment will pose serious threats to human health, inciting a series of toxic incidents. Consequently, it is very important to develop effective analytical methods. This paper presents a comprehensive review of the advancements in research pertaining to the pretreatment and detection methods in common substrates, including high performance liquid chromatography, liquid chromatography-mass spectrometry and rapid detection methods. To explore the specific sources of ATs in actual poisoning cases, the comprehensive traceability strategy based on plant morphology, chemical fingerprint analysis and DNA barcoding technology was discussed, proposing a comprehensive prospect for the development of ATs analysis and traceability, in order to provide guidance for related research within the forensic science domain.