Early identification of the use of potent benzylbenzimidazoles (nitazenes) through wastewater analysis: Two years of data from 22 countries.

Richard Bade; Dhayaalini Nadarajan; Wayne Hall; Jared Brown; Jennifer Schumann

doi:10.1111/add.70027

Early identification of the use of potent benzylbenzimidazoles (nitazenes) through wastewater analysis: Two years of data from 22 countries.

Richard Bade
, Dhayaalini Nadarajan
, Wayne Hall
, Jared Brown
,
, Jennifer Schumann

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

213 Downloads (Pure)

Abstract

Background and Aims: The use of new synthetic opioids, such as the highly potent 2-benzylbenzimidazoles (i.e. nitazene) drugs, is a global health concern because of their increased risk of fatal overdose. In the early 2020s, nitazene analogues were linked to significant numbers of overdoses in the United States. Their reach is now worldwide, with nitazene overdose deaths reported in Europe, Australia and New Zealand. The aim of this study was to measure quantities of nitazenes in wastewater samples collected from 68 locations in 22 countries, covering six continents, to understand and estimate their use.
Methods: Untreated influent wastewater samples were collected over a one-week period that included the New Year period in 2022–2023 and 2023–2024. Samples were collected from 22 countries: Australia, Austria, Belgium, Brazil, Canada, Chile, China, Cyprus, Czechia, France, Germany, Greece, Iceland, Italy, New Zealand, Nigeria, Republic of Korea, Slovenia, Spain, Sweden, United Kingdom and United States. Samples were loaded onto solid-phase extraction cartridges in the country of collection and sent to Australia for elution and analysis using sensitive liquid chromatography–mass spectrometry methods.
Results: A total of 683 individual wastewater samples were analysed across the two years:339 in 2022–2023 and 344 in 2023–2024. Two nitazene analogues—protonitazene and N-pyrrolidino etonitazene (etonitazepyne)—were found in five separate sites in the United States and Australia. In the 2022–2023 period, protonitazene was found in two sites in the United States. The following year, protonitazene was detected in two further sites in the United States, while both protonitazene and etonitazepyne were found in one site in Australia. Protonitazene mass loads ranged between 0.3 mg/day/1000 people and 100 mg/day/1000 people. Etonitazepyne was also found at mass loads between
0.2–2 mg/day/1000 people).
Conclusions: A very high mass load of protonitazene was calculated, using wastewater
analysis, for the day of 30 December 2023 in one site in Australia. Etonitazepyne
showed the same trend from a lower base. Wastewater-based nitazene surveillance
shows promise as a form of both drug early warning and ongoing monitoring of trends in
use, especially as a complementary tool to existing surveillance methods.

Original language	English
Pages (from-to)	1739-1746
Number of pages	8
Journal	Addiction
Volume	120
Issue number	9
Early online date	18 Mar 2025
DOIs	https://doi.org/10.1111/add.70027
Publication status	Published (in print/issue) - 30 Sept 2025

Bibliographical note

Publisher Copyright:
© 2025 The Author(s). Addiction published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.

Data Availability Statement

The data that support the findings of this study are available from the
corresponding author upon reasonable request.

Funding

We sincerely thank the wastewater treatment plant operators for their help in collecting samples and the support provided by the lab staff who performed the extraction of the samples. We also thank SCIEX for the use of their instrument. The Queensland Alliance for Environmental Health Sciences, The University of Queensland, gratefully acknowledges the financial support of Queensland Health, Australia. The National Centre for Youth Substance Use Research receives funding from the Commonwealth Department of Health. R.B. is the recipient of an Australian Research Council Discovery Early Career Award (project number DE220100381) funded by the Australian Government. The views expressed in this article are those of the author(s) and not necessarily those of the funding agencies. Open access publishing facilitated by The University of Queensland, as part of the Wiley \u2010 The University of Queensland agreement via the Council of Australian University Librarians.

Funders	Funder number
The University of Queensland
The University of Queensland
Australian Research Council	DE220100381

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being
SDG 6 Clean Water and Sanitation
SDG 14 Life Below Water

Keywords

early warning system
etonitazepyne,
illicit drugs,
new psychoactive substances,
rotonitazene,
wastewater-based epidemiology
illicit drugs
etonitazepyne
new psychoactive substances
protonitazene
Water Pollutants, Chemical/analysis
Benzimidazoles/analysis
Wastewater/chemistry
Humans
Europe
Substance Abuse Detection/methods
Chromatography, Liquid
Australia

Access to Document

10.1111/add.70027Licence: CC BY

Addiction - 2025 - Bade - Early identification of the use of potent benzylbenzimidazoles nitazenes through wastewaterFinal published version, 901 KBLicence: CC BY

Cite this

@article{f227bde66d06485a811d62f5ab3051c6,

title = "Early identification of the use of potent benzylbenzimidazoles (nitazenes) through wastewater analysis: Two years of data from 22 countries.",

abstract = " Background and Aims: The use of new synthetic opioids, such as the highly potent 2-benzylbenzimidazoles (i.e. nitazene) drugs, is a global health concern because of their increased risk of fatal overdose. In the early 2020s, nitazene analogues were linked to significant numbers of overdoses in the United States. Their reach is now worldwide, with nitazene overdose deaths reported in Europe, Australia and New Zealand. The aim of this study was to measure quantities of nitazenes in wastewater samples collected from 68 locations in 22 countries, covering six continents, to understand and estimate their use. Methods: Untreated influent wastewater samples were collected over a one-week period that included the New Year period in 2022–2023 and 2023–2024. Samples were collected from 22 countries: Australia, Austria, Belgium, Brazil, Canada, Chile, China, Cyprus, Czechia, France, Germany, Greece, Iceland, Italy, New Zealand, Nigeria, Republic of Korea, Slovenia, Spain, Sweden, United Kingdom and United States. Samples were loaded onto solid-phase extraction cartridges in the country of collection and sent to Australia for elution and analysis using sensitive liquid chromatography–mass spectrometry methods. Results: A total of 683 individual wastewater samples were analysed across the two years:339 in 2022–2023 and 344 in 2023–2024. Two nitazene analogues—protonitazene and N-pyrrolidino etonitazene (etonitazepyne)—were found in five separate sites in the United States and Australia. In the 2022–2023 period, protonitazene was found in two sites in the United States. The following year, protonitazene was detected in two further sites in the United States, while both protonitazene and etonitazepyne were found in one site in Australia. Protonitazene mass loads ranged between 0.3 mg/day/1000 people and 100 mg/day/1000 people. Etonitazepyne was also found at mass loads between 0.2–2 mg/day/1000 people). Conclusions: A very high mass load of protonitazene was calculated, using wastewater analysis, for the day of 30 December 2023 in one site in Australia. Etonitazepyne showed the same trend from a lower base. Wastewater-based nitazene surveillance shows promise as a form of both drug early warning and ongoing monitoring of trends in use, especially as a complementary tool to existing surveillance methods.",

keywords = "early warning system, etonitazepyne,, illicit drugs,, new psychoactive substances,, rotonitazene,, wastewater-based epidemiology, illicit drugs, etonitazepyne, new psychoactive substances, protonitazene, Water Pollutants, Chemical/analysis, Benzimidazoles/analysis, Wastewater/chemistry, Humans, Europe, Substance Abuse Detection/methods, Chromatography, Liquid, Australia",

author = "Richard Bade and Dhayaalini Nadarajan and Wayne Hall and Jared Brown and Coleman, \{H. M.\} and Jennifer Schumann",

note = "Publisher Copyright: {\textcopyright} 2025 The Author(s). Addiction published by John Wiley \& Sons Ltd on behalf of Society for the Study of Addiction.",

year = "2025",

month = sep,

day = "30",

doi = "10.1111/add.70027",

language = "English",

volume = "120",

pages = "1739--1746",

journal = "Addiction",

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T1 - Early identification of the use of potent benzylbenzimidazoles (nitazenes) through wastewater analysis: Two years of data from 22 countries.

AU - Bade, Richard

AU - Nadarajan, Dhayaalini

AU - Hall, Wayne

AU - Brown, Jared

AU - Schumann, Jennifer

A2 - Coleman, H. M.

PY - 2025/9/30

Y1 - 2025/9/30

N2 - Background and Aims: The use of new synthetic opioids, such as the highly potent 2-benzylbenzimidazoles (i.e. nitazene) drugs, is a global health concern because of their increased risk of fatal overdose. In the early 2020s, nitazene analogues were linked to significant numbers of overdoses in the United States. Their reach is now worldwide, with nitazene overdose deaths reported in Europe, Australia and New Zealand. The aim of this study was to measure quantities of nitazenes in wastewater samples collected from 68 locations in 22 countries, covering six continents, to understand and estimate their use. Methods: Untreated influent wastewater samples were collected over a one-week period that included the New Year period in 2022–2023 and 2023–2024. Samples were collected from 22 countries: Australia, Austria, Belgium, Brazil, Canada, Chile, China, Cyprus, Czechia, France, Germany, Greece, Iceland, Italy, New Zealand, Nigeria, Republic of Korea, Slovenia, Spain, Sweden, United Kingdom and United States. Samples were loaded onto solid-phase extraction cartridges in the country of collection and sent to Australia for elution and analysis using sensitive liquid chromatography–mass spectrometry methods. Results: A total of 683 individual wastewater samples were analysed across the two years:339 in 2022–2023 and 344 in 2023–2024. Two nitazene analogues—protonitazene and N-pyrrolidino etonitazene (etonitazepyne)—were found in five separate sites in the United States and Australia. In the 2022–2023 period, protonitazene was found in two sites in the United States. The following year, protonitazene was detected in two further sites in the United States, while both protonitazene and etonitazepyne were found in one site in Australia. Protonitazene mass loads ranged between 0.3 mg/day/1000 people and 100 mg/day/1000 people. Etonitazepyne was also found at mass loads between 0.2–2 mg/day/1000 people). Conclusions: A very high mass load of protonitazene was calculated, using wastewater analysis, for the day of 30 December 2023 in one site in Australia. Etonitazepyne showed the same trend from a lower base. Wastewater-based nitazene surveillance shows promise as a form of both drug early warning and ongoing monitoring of trends in use, especially as a complementary tool to existing surveillance methods.

AB - Background and Aims: The use of new synthetic opioids, such as the highly potent 2-benzylbenzimidazoles (i.e. nitazene) drugs, is a global health concern because of their increased risk of fatal overdose. In the early 2020s, nitazene analogues were linked to significant numbers of overdoses in the United States. Their reach is now worldwide, with nitazene overdose deaths reported in Europe, Australia and New Zealand. The aim of this study was to measure quantities of nitazenes in wastewater samples collected from 68 locations in 22 countries, covering six continents, to understand and estimate their use. Methods: Untreated influent wastewater samples were collected over a one-week period that included the New Year period in 2022–2023 and 2023–2024. Samples were collected from 22 countries: Australia, Austria, Belgium, Brazil, Canada, Chile, China, Cyprus, Czechia, France, Germany, Greece, Iceland, Italy, New Zealand, Nigeria, Republic of Korea, Slovenia, Spain, Sweden, United Kingdom and United States. Samples were loaded onto solid-phase extraction cartridges in the country of collection and sent to Australia for elution and analysis using sensitive liquid chromatography–mass spectrometry methods. Results: A total of 683 individual wastewater samples were analysed across the two years:339 in 2022–2023 and 344 in 2023–2024. Two nitazene analogues—protonitazene and N-pyrrolidino etonitazene (etonitazepyne)—were found in five separate sites in the United States and Australia. In the 2022–2023 period, protonitazene was found in two sites in the United States. The following year, protonitazene was detected in two further sites in the United States, while both protonitazene and etonitazepyne were found in one site in Australia. Protonitazene mass loads ranged between 0.3 mg/day/1000 people and 100 mg/day/1000 people. Etonitazepyne was also found at mass loads between 0.2–2 mg/day/1000 people). Conclusions: A very high mass load of protonitazene was calculated, using wastewater analysis, for the day of 30 December 2023 in one site in Australia. Etonitazepyne showed the same trend from a lower base. Wastewater-based nitazene surveillance shows promise as a form of both drug early warning and ongoing monitoring of trends in use, especially as a complementary tool to existing surveillance methods.

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KW - etonitazepyne,

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KW - new psychoactive substances,

KW - rotonitazene,

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KW - etonitazepyne

KW - new psychoactive substances

KW - protonitazene

KW - Water Pollutants, Chemical/analysis

KW - Benzimidazoles/analysis

KW - Wastewater/chemistry

KW - Humans

KW - Europe

KW - Substance Abuse Detection/methods

KW - Chromatography, Liquid

KW - Australia

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UR - https://www.scopus.com/pages/publications/105000819212

U2 - 10.1111/add.70027

DO - 10.1111/add.70027

M3 - Article

C2 - 40099983

SN - 0965-2140

VL - 120

SP - 1739

EP - 1746

JO - Addiction

JF - Addiction

IS - 9

ER -

Early identification of the use of potent benzylbenzimidazoles (nitazenes) through wastewater analysis: Two years of data from 22 countries.

Abstract

Bibliographical note

Data Availability Statement

Funding

UN SDGs

Keywords

Access to Document

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