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Micro-QuEChERS Extraction for Broad-Spectrum LC-QQQ-MS Quantification of Nitazene Analogs in Plasma: A Validated
Karla A O Souza1,2, Náthaly C B F Santos1,2, Rafael Lanaro1,2
1Centro de Informação e Assistência Toxicológica de Campinas (CIATox-Campinas), Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
Drug Testing and Analysis
|July 28, 2026
Summary
A new LC-MS/MS method quantifies nitazene synthetic opioids in plasma. This validated approach, using micro-QuEChERS, aids clinical and forensic toxicology by detecting emerging compounds like N-pyrrolidino protonitazene.
Area of Science:
- Clinical Toxicology
- Forensic Science
- Analytical Chemistry
Background:
- Nitazenes are potent synthetic opioids causing significant clinical and forensic toxicology concerns.
- Sensitive and selective analytical methods are crucial for detecting nitazenes in biological samples.
Purpose of the Study:
- To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying nitazene analogs and metabolites in plasma.
- To establish a reliable method for analyzing emerging synthetic opioids in clinical and forensic settings.
Main Methods:
- Utilized micro-QuEChERS (Quick, Easy, Cheap, Rugged, and Safe) for sample preparation.
- Employed LC-MS/MS for sensitive and selective quantification of nitazenes in plasma.
- Validated the method for concentration range, linearity, LOD, LLOQ, precision, and accuracy.
Main Results:
- Validated concentration range: 0.05-20 ng/mL with R² > 0.99.
- Achieved LOD of 0.025 ng/mL and LLOQ of 0.05 ng/mL.
- Demonstrated method applicability by quantifying N-pyrrolidino protonitazene in a clinical intoxication case.
Conclusions:
- The developed LC-MS/MS method with micro-QuEChERS is sensitive, selective, and suitable for targeted nitazene quantification.
- This method provides a practical approach for clinical and forensic toxicology, aiding in the identification of emerging synthetic opioid threats.
- The successful application in a real case highlights the method's utility in managing severe opioid toxicity.
