{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:18:41Z","timestamp":1790381921484,"version":"4.1.0"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"57","license":[{"start":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T00:00:00Z","timestamp":1745280000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T00:00:00Z","timestamp":1745280000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100024054","name":"Australian Criminal Intelligence Commission","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100024054","id-type":"DOI","asserted-by":"publisher"}]},{"name":"SA Health"},{"DOI":"10.13039\/501100001787","name":"University of South Australia","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001787","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Environ Sci Pollut Res"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>A class of novel synthetic opioids, nitazenes, have infiltrated the illicit drug market worldwide. Their high potency leading to overdose and fatality is a public health concern. Xylazine is a non-opiate sedative typically used for veterinary applications. The drug has recently appeared as an adulterant in the informal drug market, particularly within opioid drugs such as fentanyl and heroin. Determining the extent of use of these drugs is challenging due to low levels consumed. Wastewater analysis has been applied to monitor novel psychoactive substance use at a population level. This study presents the validation of a sensitive analytical method for the detection and quantification of trace levels of nitazenes and xylazine in wastewater. A solid phase extraction method was employed, yielding a concentration factor of 1000-fold. The method was applied to wastewater samples collected in Australia. Five different nitazenes were found near the limit of detection and quantitation levels, with detection frequencies ranging from 3 to 6% of all samples analysed. Xylazine was detected at a higher detection frequency (26% of all samples analysed). The method presented in this study could be used as an early warning system to track the use of xylazine and nitazenes internationally.<\/jats:p>","DOI":"10.1007\/s11356-025-36425-0","type":"journal-article","created":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T21:36:17Z","timestamp":1745271377000},"page":"30863-30870","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Comprehensive method to detect nitazene analogues and xylazine in wastewater"],"prefix":"10.1007","volume":"32","author":[{"given":"Emma L.","family":"Keller","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brock","family":"Peake","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bradley S.","family":"Simpson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason M.","family":"White","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9070-843X","authenticated-orcid":false,"given":"Cobus","family":"Gerber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,22]]},"reference":[{"key":"36425_CR1","doi-asserted-by":"publisher","first-page":"250","DOI":"10.1002\/dta.2484","volume":"11","author":"R Bade","year":"2019","unstructured":"Bade R, Stockham P, Painter B, Celma A, Bijlsma L, Hernandez F, White JM, Gerber C (2019) Investigating the appearance of new psychoactive substances in South Australia using wastewater and forensic data. Drug Test Anal 11:250\u2013256. https:\/\/doi.org\/10.1002\/dta.2484","journal-title":"Drug Test Anal"},{"key":"36425_CR2","doi-asserted-by":"publisher","first-page":"170781","DOI":"10.1016\/j.scitotenv.2024.170781","volume":"920","author":"R Bade","year":"2024","unstructured":"Bade R, Nadarajan D, Driver EM, Halden RU, Gerber C, Krotulski A, Hall W, Mueller JF (2024) Wastewater-based monitoring of the nitazene analogues: first detection of protonitazene in wastewater. Sci Total Environ 920:170781. https:\/\/doi.org\/10.1016\/j.scitotenv.2024.170781","journal-title":"Sci Total Environ"},{"key":"36425_CR3","doi-asserted-by":"publisher","first-page":"e2350630","DOI":"10.1001\/jamanetworkopen.2023.50630","volume":"7","author":"M Cano","year":"2024","unstructured":"Cano M, Daniulaityte R, Marsiglia F (2024) Xylazine in overdose deaths and forensic drug reports in US states, 2019\u20132022. JAMA Netw Open 7:e2350630. https:\/\/doi.org\/10.1001\/jamanetworkopen.2023.50630","journal-title":"JAMA Netw Open"},{"key":"36425_CR4","doi-asserted-by":"publisher","DOI":"10.1016\/j.focus.2024.100203","author":"C Delcher","year":"2024","unstructured":"Delcher C, Quesinberry D, Torabi S, Berry S, Keck J, Rani A, Subedi B (2024) Wastewater surveillance for xylazine in Kentucky. AJPM Focus. https:\/\/doi.org\/10.1016\/j.focus.2024.100203","journal-title":"AJPM Focus"},{"key":"36425_CR5","doi-asserted-by":"publisher","DOI":"10.1007\/s11916-024-01211-z","author":"AN Edinoff","year":"2024","unstructured":"Edinoff AN, Sall S, Upshaw WC, Spillers NJ, Vincik LY, De Witt AS, Murnane KS, Kaye AM, Kaye AD (2024) Xylazine: a drug adulterant of clinical concern. Curr Pain Headache Rep. https:\/\/doi.org\/10.1007\/s11916-024-01211-z","journal-title":"Curr Pain Headache Rep"},{"key":"36425_CR6","unstructured":"European Medicines Agency (2011) Guideline on bioanalytical method validation. https:\/\/www.ema.europa.eu\/en\/bioanalytical-method-validation-scientific-guideline. Accessed Dec 2024"},{"key":"36425_CR7","doi-asserted-by":"publisher","first-page":"109380","DOI":"10.1016\/j.drugalcdep.2022.109380","volume":"233","author":"J Friedman","year":"2022","unstructured":"Friedman J, Montero F, Bourgois P, Wahbi R, Dye D, Goodman-Meza D, Shover C (2022) Xylazine spreads across the US: a growing component of the increasingly synthetic and polysubstance overdose crisis. Drug Alcohol Depend 233:109380. https:\/\/doi.org\/10.1016\/j.drugalcdep.2022.109380","journal-title":"Drug Alcohol Depend"},{"key":"36425_CR8","doi-asserted-by":"publisher","first-page":"923","DOI":"10.1186\/s12889-024-18459-0","volume":"24","author":"H Galust","year":"2024","unstructured":"Galust H, Seltzer JA, Hardin JR, Friedman NA, Salamat J, Clark RF, Harmon J (2024) Adulterants present in the San Diego county fentanyl supply: a laboratory analysis of seized law enforcement samples. BMC Public Health 24:923. https:\/\/doi.org\/10.1186\/s12889-024-18459-0","journal-title":"BMC Public Health"},{"key":"36425_CR9","doi-asserted-by":"publisher","first-page":"e240572","DOI":"10.1001\/jamanetworkopen.2024.0572","volume":"7","author":"M Hochheimer","year":"2024","unstructured":"Hochheimer M, Strickland JC, Rabinowitz JA, Ellis JD, Dunn KE, Huhn AS (2024) Knowledge, preference, and adverse effects of xylazine among adults in substance use treatment. JAMA Netw Open 7:e240572. https:\/\/doi.org\/10.1001\/jamanetworkopen.2024.0572","journal-title":"JAMA Netw Open"},{"key":"36425_CR10","doi-asserted-by":"publisher","first-page":"468","DOI":"10.1097\/ADM.0000000000001132","volume":"17","author":"AC Holt","year":"2023","unstructured":"Holt AC, Schwope DM, Le K, Schrecker JP, Heltsley R (2023) Widespread distribution of xylazine detected throughout the United States in healthcare patient samples. J Addict Med 17:468\u2013470. https:\/\/doi.org\/10.1097\/ADM.0000000000001132","journal-title":"J Addict Med"},{"key":"36425_CR11","doi-asserted-by":"publisher","first-page":"170473","DOI":"10.1016\/j.scitotenv.2024.170473","volume":"919","author":"EL Jaunay","year":"2024","unstructured":"Jaunay EL, Bade R, Paxton KR, Nadarajan D, Barry DC, Zhai Y, Tscharke BJ, O\u2019Brien JW, Mueller J, White JM, Simpson BS, Gerber C (2024) Monitoring the use of novel psychoactive substances in Australia by wastewater-based epidemiology. Sci Total Environ 919:170473. https:\/\/doi.org\/10.1016\/j.scitotenv.2024.170473","journal-title":"Sci Total Environ"},{"key":"36425_CR12","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1093\/jat\/bkac067","volume":"46","author":"SL Kacinko","year":"2022","unstructured":"Kacinko SL, Mohr ALA, Logan BK, Barbieri EJ (2022) Xylazine: pharmacology review and prevalence and drug combinations in forensic toxicology casework. J Anal Toxicol 46:911\u2013917. https:\/\/doi.org\/10.1093\/jat\/bkac067","journal-title":"J Anal Toxicol"},{"key":"36425_CR13","doi-asserted-by":"crossref","unstructured":"Kariisa M, O\u2019Donnell J, Kumar S, Mattson CL, Goldberger BA (2023) Illicitly manufactured fentanyl\u2013involved overdose deaths with detected xylazine \u2014 United States, January 2019\u2013 June 2022, Centre for disease control and prevention. https:\/\/www.cdc.gov\/mmwr\/volumes\/72\/wr\/mm7226a4.htm. Accessed Dec 2024","DOI":"10.15585\/mmwr.mm7226a4"},{"key":"36425_CR14","doi-asserted-by":"publisher","unstructured":"Killoran S, McNamara S, Kavanagh P, O'Brien J, Lakes R (2024) Identification of N-pyrrolidino protonitazene in powders sold as heroin and associated with overdose clusters in Dublin and Cork, Ireland. Drug Test Anal 17:350\u2013357. https:\/\/doi.org\/10.1002\/dta.3707","DOI":"10.1002\/dta.3707"},{"key":"36425_CR15","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1080\/15563650.2022.2159427","volume":"61","author":"JS Love","year":"2023","unstructured":"Love JS, Levine M, Aldy K, Brent J, Krotulski AJ, Logan BK, Vargas-Torres C, Walton SE, Amaducci A, Calello D, Hendrickson R, Hughes A, Kurt A, Judge B, Pizon A, Schwarz E, Shulman J, Wiegan T, Wax P, Manini AF (2023) Opioid overdoses involving xylazine in emergency department patients: a multicenter study. Clin Toxicol (Phila) 61:173\u2013180. https:\/\/doi.org\/10.1080\/15563650.2022.2159427","journal-title":"Clin Toxicol (Phila)"},{"key":"36425_CR16","doi-asserted-by":"publisher","first-page":"182","DOI":"10.1186\/s12954-024-01097-5","volume":"21","author":"NL Michaels","year":"2024","unstructured":"Michaels NL, Bista S, Short Mejia A, Hays H, Smith GA (2024) Xylazine awareness and attitudes among people who use drugs in Ohio, 2023\u20132024. Harm Reduct J 21:182. https:\/\/doi.org\/10.1186\/s12954-024-01097-5","journal-title":"Harm Reduct J"},{"key":"36425_CR17","unstructured":"National Drug and Early Warning System (NDEWS) (2024) Alert from the NDEWS web monitoring team: kitty flipping. https:\/\/ndews.org\/novel-surveillance\/web-surveillance\/. Accessed Dec 2024"},{"key":"36425_CR18","doi-asserted-by":"publisher","DOI":"10.1111\/1742-6723.14512","author":"E Partridge","year":"2024","unstructured":"Partridge E, Stockham P, Kenneally M, Luong A, Kostakis C, Alfred S (2024) A cluster of multi-drug intoxications involving xylazine, benzimidazole opioids (nitazenes) and novel benzodiazepines in South Australia. Emerg Med Australas. https:\/\/doi.org\/10.1111\/1742-6723.14512","journal-title":"Emerg Med Australas"},{"key":"36425_CR19","doi-asserted-by":"publisher","first-page":"e40736","DOI":"10.7759\/cureus.40736","volume":"15","author":"J Pergolizzi Jr","year":"2023","unstructured":"Pergolizzi J Jr, Raffa R, LeQuang JAK, Breve F, Varrassi G (2023) Old drugs and new challenges: a narrative review of nitazenes. Cureus 15:e40736. https:\/\/doi.org\/10.7759\/cureus.40736","journal-title":"Cureus"},{"key":"36425_CR20","doi-asserted-by":"publisher","first-page":"458","DOI":"10.15585\/mmwr.mm7217a2","volume":"72","author":"E Russell","year":"2023","unstructured":"Russell E, Sisco E, Thomson A, Lopes J, Rybak M, Burnett M, Heilman D, Appley MG, Gladden M (2023) Rapid analysis of drugs: a pilot surveillance system to detect changes in the illicit drug supply to guide timely harm reduction responses \u2014 eight syringe services programs, Maryland, November 2021\u2013August 2022. MMWR Morb Mortal Wkly Rep 72:458\u2013462","journal-title":"MMWR Morb Mortal Wkly Rep"},{"key":"36425_CR21","doi-asserted-by":"publisher","first-page":"e6","DOI":"10.1093\/jat\/bkac062","volume":"47","author":"JL Schumann","year":"2023","unstructured":"Schumann JL, Syrjanen R, Alford K, Mashetty S, Castle JW, Rotella J, Maplesden J, Greene SL (2023) Intoxications in an Australian emergency department involving \u2018nitazene\u2019 benzylbenzimidazole synthetic opioids (etodesnitazene, butonitazene and protonitazene). J Anal Toxicol 47:e6\u2013e9. https:\/\/doi.org\/10.1093\/jat\/bkac062","journal-title":"J Anal Toxicol"},{"key":"36425_CR22","unstructured":"Tanz LJ, Stewart A, Gladden RM, Ko JY, Owens L, O\u2019Donnell J (2024) Detection of illegally manufactured fentanyls and carfentanil in drug overdose deaths \u2014 United States, 2021\u2013 2024, Centers for disease control and prevention U.S. Department of Health and Human Services.\nhttps:\/\/www.cdc.gov\/mmwr\/volumes\/73\/wr\/mm7348a2.htm#:~:text=During%20January%202021%E2%80%93June%202024%2C%20IMFs%20were%20detected%20in%2081.5,time. Accessed Dec 2024"},{"key":"36425_CR23","unstructured":"The National Centre for Clinical Research on Emerging Drugs (2024) Emerging drug briefing: increasing reports of nitazene toxicity in Australia.\nhttps:\/\/nccred.org.au\/resources\/emerging-drug-briefing-nitazenes. Accessed Dec 2024"},{"key":"36425_CR24","doi-asserted-by":"publisher","first-page":"1072","DOI":"10.1021\/acschemneuro.1c00037","volume":"12","author":"I Ujvary","year":"2021","unstructured":"Ujvary I, Christie R, Evans-Brown M, Gallegos A, Jorge R, de Morais J, Sedefov R (2021) DARK classics in chemical neuroscience: etonitazene and related benzimidazoles. ACS Chem Neurosci 12:1072\u20131092. https:\/\/doi.org\/10.1021\/acschemneuro.1c00037","journal-title":"ACS Chem Neurosci"},{"key":"36425_CR25","unstructured":"UNODC (2024): Early warning advisory on new psychoactive substances, UNODC"},{"key":"36425_CR26","unstructured":"US Department of Health and Human Services Food and Drug Administration (2018) Bioanalytical method validation: guidance for industry.\nhttps:\/\/www.fda.gov\/files\/drugs\/published\/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf. Accessed Dec 2024"},{"key":"36425_CR27","unstructured":"U.S. Department of Justice: Drug Enforcement Administration (2022) The growing threat of xylazine and its mixture with illicit drugs. https:\/\/www.dea.gov\/sites\/default\/files\/2022-12\/The%20Growing%20Threat%20of%20Xylazine%20and%20its%20Mixture%20with%20Illicit%20Drugs.pdf. Accessed Apr 2024"}],"container-title":["Environmental Science and Pollution Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11356-025-36425-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s11356-025-36425-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11356-025-36425-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T06:23:29Z","timestamp":1768544609000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s11356-025-36425-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,22]]},"references-count":27,"journal-issue":{"issue":"57","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["36425"],"URL":"https:\/\/doi.org\/10.1007\/s11356-025-36425-0","relation":{},"ISSN":["1614-7499"],"issn-type":[{"value":"1614-7499","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,22]]},"assertion":[{"value":"7 January 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 April 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 April 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"No ethical approval is required for this study.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"This is not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to participate"}},{"value":"This is not applicable.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}