Advances and Challenges in LSD Detection: Analytical Techniques, Matrix Selection, and Validation Gaps in Forensic Toxicology

Detecting LSD in biological samples is difficult because the drug is taken in very small amounts, breaks down quickly in the body, and resembles other similar compounds. This review examined published analytical methods from 1978 to 2025 for measuring LSD and its metabolites in blood, urine, hair, oral fluid, and vitreous humor. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the most sensitive and widely validated technique, but there are inconsistencies in how methods are validated for different sample types and in how they account for sample degradation. Few studies meet modern forensic validation standards, which limits the reliability of results in legal cases. The review identifies a need for standardized, rapid, and environmentally friendly methods that can detect LSD and its analogs at extremely low levels.

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Abstract

Lysergic acid diethylamide (LSD) remains a significant forensic and public health concern due to its widespread abuse and association with drug-facilitated crimes. Detecting LSD and its analogs in biological specimens, particularly postmortem matrices, presents analytical challenges stemming from its low dosage, rapid metabolism, and structural similarities to novel lysergamides. This review critically examines trends in validated analytical methods for LSD detection in forensic toxicology. A systematic review of literature from 1978 to 2025 was conducted using databases, such as PubMed, ScienceDirect, and Google Scholar. The analysis focused on reported methodologies for LSD and its metabolites across various matrices, including blood, urine, hair, oral fluid, and vitreous humor. Extraction techniques (LLE, SPE, DLLME) and analytical platforms (GC-MS/MS, LC-MS/MS, CE-MS) were compared, with emphasis on validation parameters, such as sensitivity, specificity, recovery, LOD, LOQ, matrix effects, and stability. The review identifies LC-MS/MS as the most sensitive and widely validated technique; however, discrepancies remain in matrix-specific validations and stability assessments. Challenges include the lack of certified reference materials for LSD analogs, matrix-dependent degradation, and limited methods for emerging sample types, such as dried blood spots (DBS). Few studies fully comply with modern forensic validation guidelines, limiting the reproducibility and admissibility of results in legal settings. This review highlights critical gaps in current forensic LSD detection protocols and underscores the need for standardized, validated methods applicable to diverse matrices. Future research should prioritize the development of rapid, eco-friendly, high-throughput methods capable of detecting LSD and its analogs at ultra-trace levels.