Analysis of stimulant drugs in the wastewater of five Nordic capitals

Publisher's version (útgefin grein)Wastewater-based epidemiology is an efficient way to assess illicit drug use, complementing currently used methods retrieved from different data sources. The aim of this study is to compare stimulant drug use in five Nordic capital cities that include for the first time wastewater samples from Torshavn in the Faroe Islands. Currently there are no published reports that compare stimulant drug use in these Nordic capitals. All wastewater samples were analyzed using solid phase extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry. The results were compared with data published by the European Monitoring Centre for Drugs and Drug Addiction based on illicit drugs in wastewater from over 50 European cities. Confirming previous reports, the results showed high amphetamine loads compared with other European countries. Very little apparent abuse of stimulant drugs was detected in Torshavn. Methamphetamine loads were the highest from Helsinki of the Nordic countries, indicating substantial fluctuations in the availability of the drug compared with previous studies. Methamphetamine loads from Oslo confirmed that the use continues to be high. Estimated cocaine use was found to be in the lower range compared with other cities in the southern and western part of Europe. Ecstasy and cocaine showed clear variations between weekdays and weekends, indicating recreational use. This study further demonstrates geographical trends in the stimulant drug market in five Nordic capitals, which enables a better comparison with other areas of the continent.We wish to acknowledgesupport by COST (EuropeanCooperation inScience and Technology) [Grant number ES1307], as well as the Icelan-dic Research Fund [Grant number 163049-051] and the University ofIceland Research Fund [Grant number HI17060092].Peer Reviewe

Three years of wastewater surveillance for new psychoactive substances from 16 countries

The proliferation of new psychoactive substances (NPS) over recent years has made their surveillance complex. The analysis of raw municipal influent wastewater can allow a broader insight into community consumption patterns of NPS. This study examines data from an international wastewater surveillance program that collected and analysed influent wastewater samples from up to 47 sites in 16 countries between 2019 and 2022. Influent wastewater samples were collected over the New Year period and analysed using validated liquid chromatography - mass spectrometry methods. Over the three years, a total of 18 NPS were found in at least one site. Synthetic cathinones were the most found class followed by phenethylamines and designer benzodiazepines. Furthermore, two ketamine analogues, one plant based NPS (mitragynine) and methiopropamine were also quantified across the three years. This work demonstrates that NPS are used across different continents and countries with the use of some more evident in particular regions. For example, mitragynine has highest mass loads in sites in the United States, while eutylone and 3-methylmethcathinone increased considerably in New Zealand and in several European countries, respectively. Moreover, 2F-deschloroketamine, an analogue of ketamine, has emerged more recently and could be quantified in several sites, including one in China, where it is considered as one of the drugs of most concern. Finally, some NPS were detected in specific regions during the initial sampling campaigns and spread to additional sites by the third campaign. Hence, wastewater surveillance can provide an insight into temporal and spatial trends of NPS use

Fíkniefni í frárennsli frá Reykjavík

Mælingar á fíkniefnum og lyfseðilsskyldum lyfjum í frárennslisvatni hafa undanfarin ár verið notaðar til þess að meta notkun efnanna. Aðferðafræðin byggir á þeirri kenningu að hægt sé að líta á frárennslisvatn sem samansafn þvagsýna frá heilu samfélagi. Áreiðanlegar niðurstöður eru fengnar á mjög fljótvirkan hátt án þess að inngripum sé beitt. Aðal markmið þessarar rannsóknar var að setja upp og gilda greiningaraðferð fyrir fíkniefni og lyfseðilsskyld lyf í frárennsli frá Reykjavík. Fastfasa súluskiljun (SPE) var notuð við úrhlutun fíkniefna og lyfja í frárennslisvatni frá Reykjavík með ásættanlegum heimtum. Háþrýstivökvagreinir tengdur tvöföldum massaskynjara (UPLC-MS/MS) var notaður við greiningu efnanna. Frárennslissýnum var safnað í Reykjavík á ellefu tímapunktum frá febrúar 2017 til júní 2020. Algeng fíkniefni sem notuð eru á Íslandi voru valin til greiningar ásamt lyfseðilsskyldum lyfjum með þekkta misnotkun. Notkun efnanna var metin í mg/dag/1000 íbúa. Langtímaþróun í fíkniefnanotkun í Reykjavik var metin með mælingum á frárennslisvatni. Notkun kókaíns hækkaði verulega frá 2017 til 2019 en hafði minnkað umtalsvert í júní 2020 á tímum kórónuveirunnar. Notkun bæði amfetamíns og metamfetamíns sýndu merki aukningar frá 2017 til 2020, en notkun amfetamíns var mun meiri en metamfetamíns. Notkun 3,4 methylene-dioxymethamphetamine (MDMA) var stöðug frá 2017 til 2020. Notkun kannabis var stöðug frá 2017 til 2019 en sýndi merki aukningar á tímum kórónuveirunnar árið 2020. Aukning í notkun kókaíns og MDMA sást einnig um helgar þegar borið var saman við aðra vikudaga og einnig á tónlistarhátíð sem haldin var í Reykjavík. Niðurstöður byggðar á mælingum á frárennslisvatni voru bornar saman við aðra vísa að fíkniefnanotkun, fjölda mála vegna fíkniefnaaksturs og gögn um haldlagt magn fíkniefna. Afkastamikil sýnameðhöndlunaraðferð var þróuð þar sem minna rúmmál frárennslissýna var úrhlutað samanborið við aðrar hefðbundnari aðferðir. Þessi aðferð var þróuð bæði fyrir fíkniefni og lyfseðilsskyld lyf. Mikill tímasparnaður felst í því að úrhluta minna rúmmál af sýni, samanborið við aðrar eldri aðferðir og fást þá niðurstöður á mun skemmri tíma. Frárennslisýni frá fjórum höfuðborgum á Norðurlöndum (Osló, Stokkhólmur, Reykjavík og Þórshöfn) voru úrhlutuð með þessari aðferð. Gögn frá Helsinki voru einnig fengin og þróun í notkun örvandi fíkniefna var metin milli borganna. Niðurstöður sýndu að amfetamínmagn í frárennsli var mest í Reykjavík og Stokkhólmi. Magn metamfetamíns í frárennsli var hæst í Helsinki og magn MDMA var mest í Osló. Magn kókaíns í frárennsli var sambærilegt í Osló, Stokkhólmi og Reykjavík en mun lægra í Helsinki. Augljós aukning í magni MDMA og kókaíns sást um helgar sem bendir til að notkun efnanna til afþreyingar sé vinsæl á Norðurlöndum. Þróun í notkun fíkniefna og lyfja var metin með mælingum á frárennsli með góðum árangri ásamt samanburði við önnur lönd. Niðurstöður byggðar á mælingum á frárennslisvatni voru almennt sambærilegar gögnum yfir fíkniefnaakstur og haldlagningar. Hægt er að ná fram heildstæðari mynd af notkun fíkniefna í Reykjavík með því að sameina niðurstöður sem byggðar eru á mælingum á frárennsli og önnur gögn. Þetta getur veitt hagsmunaaðilum eins og lögregluyfirvöldum og heilbrigðisstarfsmönnum mikilvægar upplýsingar sem nota má í baráttunni gegn fíkniefnavandanum.Wastewater-based epidemiology (WBE) has in recent years emerged as a reliable way to estimate the use of illicit drugs and pharmaceuticals at a community level. This methodology is based on the hypothesis that wastewater can be considered a pooled urine sample of a total population. This is a non-intrusive tool which provides objective results in near real-time. The primary aim of this project was to adapt and validate a reliable analytical method for illicit drugs and pharmaceuticals in wastewater from Reykjavik. A solid phase extraction (SPE) method was adapted for the analysis of illicit drugs and pharmaceuticals in wastewater samples from Reykjavik with satisfactory recoveries. Ultra-performance liquid chromatography coupled to a tandem mass spectrometry (UPLC-MS/MS) was used for the analysis of samples. Wastewater samples were collected in Reykjavik at eleven timepoints from February 2017 to June 2020. The estimated use of the analytes of interest was expressed as mg/day/1000 inhabitants. Temporal trends in illicit drug use in Reykjavik were successfully assessed using WBE. The results showed that cocaine use had increased extensively from 2017 to 2019 but had decreased considerably in June 2020 during the COVID-19 pandemic. Amphetamine and methamphetamine both showed signs of increased use between 2017 and 2020, but amphetamine use remained significantly higher. MDMA use was estimated to be stable from 2017 to 2020. Estimated cannabis use by WBE was stable from 2017 to 2019 but showed signs of an increase in 2020 during the pandemic. Trends between weekdays and during special events were also observed showing a significant increase in cocaine and MDMA use during weekends and a music festival held in Reykjavik. Further, a comparison was made with other indicators of drug use based on seizure data and driving under the influence cases. A high-throughput SPE micro-extraction method combined with large volume injection and post-loop mixing was developed for both illicit drugs and pharmaceuticals. By extracting smaller volumes of samples compared to other conventional methods, the time per sample was lowered considerably allowing for faster reporting of results. Samples from four Nordic capitals (Oslo, Stockholm, Reykjavik, and Torshavn) were analysed using SPE micro-extraction where spatial trends were successfully estimated alongside data obtained from Helsinki. The results showed high amphetamine loads in Reykjavik and Stockholm. Methamphetamine loads were highest in Helsinki and MDMA loads were highest from Oslo. Cocaine loads were similar from Oslo, Stockholm, and Reykjavik, but notably lower from Helsinki. A clear increase was observed in MDMA and cocaine loads during weekends indicating recreational use of the drugs. Both temporal and spatial trends in illicit drug use were successfully estimated by using WBE. Overall, results by WBE were comparable with the other indicators of drug use. By complimenting data by WBE and other methods used to assess the use of drugs, a more comprehensive picture of the community drug abuse can be achieved. This could provide stakeholders such as the law enforcement or health care authorities valuable and useful information in their fight against drug abuse.Icelandic Research Fund and the University of Iceland Research Fun

Þróun á LC-MS/MS aðferð til magngreiningar á kortisóli og kortisóni

Cortisol is one of the most important hormones in the body. Deficiency or abundance of this hormone can cause serious effects. The enzyme 11βHSD2 is an important enzyme in the metabolism of the active hormone cortisol. 11βHSD2 converts the active hormone cortisol to the inactive hormone cortisone and thereby prevents localized over- or under-expression in the body tissue. This conversion is inhibited by glycyrrhetinic acid along with other chemicals which can induce hypertension, hypokalaemia and other diseases. When searching for the cause of decreased metabolism of cortisol it is important to be able to measure the activity of 11βHSD2. Studies have shown that measurement of the ratio between cortisol and cortisone can be used to evaluate the enzymes activity. The aim of this study was to develop a more specific high through-put method based on LC-MS/MS for simultaneous quantification of cortisol, cortisone and glycyrrhetinic acid. Quantification method for all analytes was optimized with a chemometric approach. ESI ionization source in positive ion-mode was used. D-optimal design was utilized in the experimental screening of parameters. CCF-design was utilized in the optimization of the method. SPE and protein precipitation were tested for the sample preparation of urine and plasma samples. Experimental screening indicated a quadratic system where collision energy and organic phase had the most effect on the responses for both cortisol and cortisone but glycyrrhetinic acid was eliminated from the model due to poor chromatography. The method was optimized on both HPLC-MS/MS and UPLC-MS/MS where the result in both cases was that collision energy had the most significant effect on the responses of all analytes. A short method validation was performed where LOQ was determined to be 5ng/ml. Recovery and matrix effect showed good results except for glycyrrhetinic acid which showed ion-enhancement. After excluding one defected sample, the accuracy and precision of cortisol and cortisone met the acceptance criteria.Kortisól er eitt mikilvægasta hormón líkamans og getur skortur eða ofgnótt þess haft alvarleg áhrif á líkamsstarfssemina. Ensímið 11βHSD2 gegnir mikilvægu hlutverki í niðurbroti virka hormónsins kortisól. Með niðurbroti kortisóls í hið óvirka hormón kortisón, varnar ensímið staðbundinni ofvirkni þess eða ofvirkni í vefjum líkamans. Ýmis efni eru þekkt fyrir að geta hindrað virkni 11βHSD2 eins og t.d. lakkríssýra, sem getur leitt til ofþrýstings, kalíumskorts eða annarra sjúkdóma. Þegar leitað er að orsakavaldi minnkaðs niðurbrots á kortisóli er mikilvægt að geta mælt virkni þessa ensíms, 11βHSD2. Rannsóknir hafa sýnt að mæling á hlutfalli útskilnaðar á fríu kortisóli og kortisóni kemst næst því að mæla virkni ensímsins. Markmið þessa rannsóknarverkefnis er að þróa sérhæfða og afkastamikla aðferð byggða á LC-MS/MS fyrir greiningu á kortisóli, kortisóni og lakkríssýru. Magngreiningaraðferð fyrir kortisól, kortisón og lakkríssýru var hámörkuð með chemometrics nálgun og var jákvæð rafúðunarjónun (ESI) notuð við þróun aðferðarinnar. D-optimal hönnun var notuð í skimun á marktækt mikilvægum breytum sem hafa áhrif á næmni kórtisóls og kortisóns. CCF-hönnun var notuð við hámörkun aðferðarinnar. Sýnameðhöndlunaraðferðirnar fastfasa súluskiljun (SPE) og próteinfelling voru þróaðar á þvag og plasma sýnum. Skimun gaf til kynna að kvaðratískt kerfi liggi að baki. Árekstrarorka og tegund lífræns leysis hafði mest áhrif á næmni kortisóls og kortisóns. Lakkríssýra var útilokuð úr líkaninu þar sem léleg litskiljun kom í veg fyrir að hægt væri að leggja mat á toppa. Hámörkun á bæði HPLC-MS/MS og UPLC-MS/MS þar sem árekstrarorka reyndist hafa mest áhrif á næmni kortisóls og kortisóns. Stutt gilding var framkvæmd þar sem kom í ljós að lægsti mælanlegi styrkur kortisóls og kortisóns var 5ng/ml. Endurheimt og matrixu áhrif fyrir kortisól og kortisón voru innan leyfilegra marka en lakkríssýra sýndi jónunar aukningu. Að undanskildu einu gölluðu sýni féll nákvæmni og áreiðanleiki aðferðarinnar innan leyfilegra marka fyrir kortisól og kortisón

Illicit drug use in Reykjavik by wastewater-based epidemiology

Publisher Copyright: © 2021 The Authors We want to acknowledge Birgir Tómas Arnar from Verkís hf. and Veitur ohf., Reykjavik Iceland, for their valuable cooperation in providing technical information and assisting with sample collection. We want to thank the DPT and the National Commissioner of the Icelandic Police for providing important data. We also want to acknowledge the Icelandic Research Fund [Grant number 163049-051] and the University of Iceland Research Fund for their contributions.Estimation of illicit drug use on a community level by wastewater-based epidemiology (WBE) is both an objective and reliable way to establish near real-time results. Wastewater samples were collected at eleven timepoints in Reykjavik from 2017 to 2020. The use of commonly abused illicit drugs in Iceland (amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), cocaine, and cannabis) was estimated. Solid phase extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry was used for analysis. Estimated amphetamine and methamphetamine use showed signs of an increase from 2017 to 2020 with amphetamine being the dominant stimulant on the market. MDMA use remained stable from 2017 to 2020. Results showed a large increase in cocaine use from 2017 to 2019 but interestingly, a marked decrease in 2020 during the COVID-19 pandemic. Cannabis use was stable from 2017 to 2019 but showed signs of an increase during the pandemic in 2020. Results by WBE corresponded with data based on two other indicators of drug use, seizure data and driving under the influence cases. Both temporal and spatial trends in illicit drug use were successfully estimated by using WBE, complimenting other indicators which provided a comprehensive picture of drug abuse in Reykjavik.Peer reviewe

Pharmaceuticals in Northern environments; what, where and how much?

The present study has investigated and collected sales statistics of pharmaceuticals in Sweden, Norway and Iceland. A literature review of waste water treatment plants (WWTP) and the efficiency of different WWTP techniques was conducted and combined with a literature review of metabolization of pharmaceuticals in humans. These data have been linked and used for calculations of the amount of pharmaceuticals and their metabolites that actually reaches the aquatic recipient. A future recommendation is to include metabolites of pharmaceuticals in environmental impact studies to be able to assess and account for the chemicals that actually pass through a WWTP to the aquatic environment. No large differences regarding pharmaceuticals used were observed, and many were common in both Sweden, Iceland and Norway

A high-throughput solid-phase microextraction and post-loop mixing large volume injection method for water samples

This article presents a novel approach for the analysis of 13 drugs in wastewater for use in wastewater-based epidemiology (WBE) studies. Sample preparation remains one of the principal bottlenecks in modern high-throughput analysis by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The proposed methodology is based on the micro-extraction of small volumes (1 ml) of wastewater using a HLB 96-well microplate and both large volume injection (LVI) and post-loop mixing injection (PLM). With this configuration, the limits of quantification (LOQ) were below the reported environmental concentrations of the target compounds in wastewater. Furthermore, both the complexity of collecting, transporting and storing the wastewater sample, sample preparation time, cost and amount of solvent used are all diminished, enhancing the suitability of this methodology for future WBE studies. A new workflow is also proposed in order to create a virtual specimen library bank for WBE by using high-resolution mass spectrometry (HRMS). The method was validated and the limits of quantification were between 0.2 and 6.3 ng L. The relative standard deviations (RSD) for a standard mixture at 200 ng L (n = 6) was between 3.4 and 14.4% while the recoveries for the 13 drug target residues (DTR) were between 92 and 110%. The developed and validated method was finally successfully applied to 10 wastewater samples collected from Oslo, Norway

Multi-year inter-laboratory exercises for the analysis of illicit drugs and metabolites in wastewater: development of a quality control system

Thirty-seven laboratories from 25 countries present the development of an inter-laboratory testing scheme for the analysis of seven illicit drug residues in standard solutions, tap- and wastewater. Almost 10 000 concentration values were evaluated: triplicates of up to five samples and 26 laboratories per year. The setup was substantially improved with experiences gained across the six repetitions (e.g. matrix type, sample conditions, spiking levels). From this, (pre-)analytical issues (e.g. pH adjustment, filtration) were revealed for specific analytes which resulted in formulation of best-practice protocols for inter-laboratory setup and analytical procedures. The results illustrate the effectiveness of the inter-laboratory setup to assess laboratory performance in the framework of wastewater-based epidemiology. The exercise proved that measurements of laboratories were of high quality (>80% satisfactory results for six out of seven analytes) and that analytical follow-up is important to assist laboratories in improving robustness of wastewater-based epidemiology results

Spatio‐temporal assessment of illicit drug use at large scale: evidence from 7 years of international wastewater monitoring

Background and aims Wastewater‐based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population‐normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011–17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data. Design Analysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017. Setting and Participants Catchment areas of 143 wastewater treatment plants in 120 cities in 37 countries. Measurements Parent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ9‐tetrahydrocannabinol (11‐nor‐9‐carboxy‐Δ9‐tetrahydrocannabinol) were measured in wastewater using liquid chromatography–tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world‐wide, and temporal trends were discerned at European level by comparing 2011–13 drug loads versus 2014–17 loads. Findings Benzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North–Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge. Conclusions The analysis of wastewater to quantify drug loads provides near real‐time drug use estimates that globally correspond to prevalence and seizure data