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Testing for protonitazene in human hair using LC-MS-MS.

Pascal Kintz^1,2, Alice Ameline¹, Laurie Gheddar¹

¹Laboratoire de toxicologie, Institut de médecine légale, Strasbourg 67000, France.

Journal of Analytical Toxicology

|June 5, 2024

Summary

A new method detects protonitazene, a potent synthetic opioid, in human hair. This breakthrough allows identification in overdose cases and living individuals, addressing the growing public health threat of nitazenes.

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Area of Science:

Forensic Toxicology
Analytical Chemistry
Pharmacology

Background:

Protonitazene, a synthetic benzimidazole opioid from the nitazene class, was developed in the 1950s as an analgesic but was never marketed due to safety concerns.
Despite its resurgence as a novel psychoactive substance since 2019, protonitazene detection in human hair has not been previously reported.

Purpose of the Study:

To develop and validate a specific analytical procedure for identifying protonitazene in human hair specimens.
To establish a reliable method for detecting protonitazene in post-mortem and living individuals to address its increasing prevalence.

Main Methods:

Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for protonitazene identification.
Hair samples were incubated in a borate buffer (pH 9.5) with fentanyl-d5 as an internal standard, followed by organic solvent extraction.
Chromatographic separation was achieved using an HSS C18 column with a 15-minute gradient elution.

Main Results:

The validated method demonstrated linearity from 1 to 100 pg/mg with a limit of detection of 0.1 pg/mg.
No interference was observed from a wide range of other opioids, fentanyl derivatives, and novel synthetic opioids.
Protonitazene was detected in hair samples from two deceased individuals (70 and >7600 pg/mg) and two living individuals (14 and 54 pg/mg).

Conclusions:

The developed LC-MS/MS method is specific and sensitive for protonitazene detection in human hair.
This method provides a crucial tool for forensic investigations involving nitazene abuse.
The findings highlight the urgent need for monitoring and addressing the public health risks associated with nitazenes.