Malcolm J. Reid

Comparing illicit drug use in 19 European cities through sewage analysis.

The analysis of sewage for urinary biomarkers of illicit drugs is a promising and complementary approach for estimating the use of these substances in the general population. For the first time, this approach was simultaneously applied in 19 European cities, making it possible to directly compare illicit drug loads in Europe over a 1-week period. An inter-laboratory comparison study was performed to evaluate the analytical performance of the participating laboratories. Raw 24-hour composite sewage samples were collected from 19 European cities during a single week in March 2011 and analyzed for the urinary biomarkers of cocaine, amphetamine, ecstasy, methamphetamine and cannabis using in-house optimized and validated analytical methods. The load of each substance used in each city was back-calculated from the measured concentrations. The data show distinct temporal and spatial patterns in drug use across Europe. Cocaine use was higher in Western and Central Europe and lower in Northern and Eastern Europe. The extrapolated total daily use of cocaine in Europe during the study period was equivalent to 356 kg/day. High per capita ecstasy loads were observed in Dutch cities, as well as in Antwerp and London. In general, cocaine and ecstasy loads were significantly elevated during the weekend compared to weekdays. Per-capita loads of methamphetamine were highest in Helsinki and Turku, Oslo and Budweis, while the per capita loads of cannabis were similar throughout Europe. This study shows that a standardized analysis for illicit drug urinary biomarkers in sewage can be applied to estimate and compare the use of these substances at local and international scales. This approach has the potential to deliver important information on drug markets (supply indicator).

Spatial differences and temporal changes in illicit drug use in Europe quantified by wastewater analysis

Aims To perform wastewater analyses to assess spatial differences and temporal changes of illicit drug use in a large European population. Design Analyses of raw wastewater over a 1-week period in 2012 and 2013. Setting and Participants Catchment areas of wastewater treatment plants (WWTPs) across Europe, as follows: 2012: 25 WWTPs in 11 countries (23 cities, total population 11.50 million); 2013: 47 WWTPs in 21 countries (42 cities, total population 24.74 million). Measurements Excretion products of five illicit drugs (cocaine, amphetamine, ecstasy, methamphetamine, cannabis) were quantified in wastewater samples using methods based on liquid chromatography coupled to mass spectrometry. Findings Spatial differences were assessed and confirmed to vary greatly across European metropolitan areas. In general, results were in agreement with traditional surveillance data, where available. While temporal changes were substantial in individual cities and years (P ranging from insignificant to <10−3), overall means were relatively stable. The overall mean of methamphetamine was an exception (apparent decline in 2012), as it was influenced mainly by four cities. Conclusions Wastewater analysis performed across Europe provides complementary evidence on illicit drug consumption and generally concurs with traditional surveillance data. Wastewater analysis can measure total illicit drug use more quickly and regularly than is the current norm for national surveys, and creates estimates where such data does not exist.

Evaluation of Uncertainties Associated with the Determination of Community Drug Use through the Measurement of Sewage Drug Biomarkers

The aim of this study was to integrally address the uncertainty associated with all the steps used to estimate community drug consumption through the chemical analysis of sewage biomarkers of illicit drugs. Uncertainty has been evaluated for sampling, chemical analysis, stability of drug biomarkers in sewage, back-calculation of drug use (specific case of cocaine), and estimation of population size in a catchment using data collected from a recent Europe-wide investigation and from the available literature. The quality of sampling protocols and analytical measurements has been evaluated by analyzing standardized questionnaires collected from 19 sewage treatments plants (STPs) and the results of an interlaboratory study (ILS), respectively. Extensive reviews of the available literature have been used to evaluate stability of drug biomarkers in sewage and the uncertainty related to back-calculation of cocaine use. Different methods for estimating population size in a catchment have been compared and the variability among the collected data was very high (7-55%). A reasonable strategy to reduce uncertainty was therefore to choose the most reliable estimation case by case. In the other cases, the highest uncertainties are related to the analysis of sewage drug biomarkers (uncertainty as relative standard deviation; RSD: 6-26% from ILS) and to the back-calculation of cocaine use (uncertainty; RSD: 26%). Uncertainty can be kept below 10% in the remaining steps, if specific requirements outlined in this work are considered. For each step, a best practice protocol has been suggested and discussed to reduce and keep to a minimum the uncertainty of the entire procedure and to improve the reliability of the estimates of drug use.

In Situ Calibration of a Passive Sampling Device for Selected Illicit Drugs and Their Metabolites in Wastewater, And Subsequent Year-Long Assessment of Community Drug Usage

Polar organic chemical integrative samplers (POCIS) were calibrated in situ for selected illicit drugs and their metabolites at a sewage treatment works. Eleven out of 13 target compounds were detected and eight of those exhibited linear uptake kinetics with sampling rates between 0.035 and 0.150 L d(-1). Subsequently POCIS were deployed for 2 week periods over the course of a whole year, in order to examine trends in drug usage. Amphetamine and methamphetamine showed several similar peaks in concentration during the course of the year as did cocaine and two of its metabolites. Low levels of ecstasy were observed, with a prominent peak in May and a steady increase toward the end of the year. The antihistamine Cetirizine showed a clear increase in use during the summer months as expected and back calculation of the yearly dosage from POCIS accumulations yielded very similar results to that registered in the Norwegian prescription database. Estimations of cocaine usage using the parent compound averaged between 0.31 and 2.8 g d(-1) per 1000 inhabitants. POCIS is a cost-effective technique for the long-term monitoring of drug usage of a defined population and may overcome the difficulties of representative sampling associated with autosampling equipment.

Environmental occurrence and risk of organic UV filters and stabilizers in multiple matrices in Norway

Eight organic UV filters and stabilizers were quantitatively determined in wastewater sludge and effluent, landfill leachate, sediments, and marine and freshwater biota. Crab, prawn and cod from Oslofjord, and perch, whitefish and burbot from Lake Mjøsa were selected in order to evaluate the potential for trophic accumulation. All of the cod livers analysed were contaminated with at least 1 UV filter, and a maximum concentration of almost 12 μg/g wet weight for octocrylene (OC) was measured in one individual. 80% of the cod livers contained OC, and approximately 50% of cod liver and prawn samples contained benzophenone (BP3). Lower concentrations and detection frequencies were observed in freshwater species and the data of most interest is the 4 individual whitefish that contained both BP3 and ethylhexylmethoxycinnamate (EHMC) with maximum concentrations of almost 200 ng/g wet weight. The data shows a difference in the loads of UV filters entering receiving water dependent on the extent of wastewater treatment. Primary screening alone is insufficient for the removal of selected UV filters (BP3, Padimate, EHMC, OC, UV-234, UV-327, UV-328, UV-329). Likely due in part to the hydrophobic nature of the majority of the UV filters studied, particulate loading and organic carbon content appear to be related to concentrations of UV filters in landfill leachate and an order of magnitude difference in these parameters correlates with an order of magnitude difference in the effluent concentrations of selected UV filters (Fig. 2). From the data, it is possible that under certain low flow conditions selected organic UV filters may pose a risk to surface waters but under the present conditions the risk is low, but some UV filters will potentially accumulate through the trophic food chain.

Analysis and Interpretation of Specific Ethanol Metabolites, Ethyl Sulfate, and Ethyl Glucuronide in Sewage Effluent for the Quantitative Measurement of Regional Alcohol Consumption

BACKGROUND The quantitative measurement of urinary metabolites in sewage streams and the subsequent estimation of consumption rates of the parent compounds have previously been demonstrated for pharmaceuticals and narcotics. Ethyl sulfate and ethyl glucuronide are excreted in urine following the ingestion of alcohol, and are useful biomarkers for the identification of acute alcohol consumption. This study reports a novel ion-exchange-mediated chromatographic method for the quantitative measurement of ethyl sulfate and ethyl glucuronide in sewage effluent, and presents a novel calculation method for the purposes of relating the resulting sewage concentrations with rates of alcohol consumption in the region. METHODS A total of 100 sewage samples covering a 25-day period were collected from a treatment plant servicing approximately 500,000 people, and analyzed for levels of ethyl sulfate and ethyl glucuronide. The resulting data were then used to estimate combined alcohol consumption rates for the region, and the results were compared with alcohol related sales statistics for the same region. RESULTS Ethyl glucuronide was found to be unstable in sewage effluent. Ethyl sulfate was stable and measurable in all samples at concentrations ranging from 16 to 246 nM. The highest concentrations of the alcohol biomarker were observed during weekend periods. Sixty one percent of the total mass of ethyl sulfate in sewage effluent corresponds to alcohol consumption on Friday and Saturday. Sales statistics for alcohol show that consumption in the region is approximately 6,750 kg/d. The quantity of ethyl sulfate passing through the sewage system is consistent with consumption of 4,900 to 7,800 kg/d. CONCLUSIONS   Sewage epidemiology assessments of ethyl sulfate can provide accurate estimates of community alcohol consumption, and detailed examination of the kinetics of this biomarker in sewage streams can also identify time-dependent trends in alcohol consumption patterns.

Quantitative assessment of time dependent drug-use trends by the analysis of drugs and related metabolites in raw sewage

BACKGROUND Accurate and timely information on the scale and dynamics of drug consumption is important for assessing the needs of law enforcement and public health services in a community. AIMS This paper presents a detailed examination of a comprehensive sewage-sampling campaign for the purposes of increasing an understanding of the dynamics of drug-flows in sewage streams, and developing new methodology by which this technique can support traditional drug-use surveys. METHODS A total of 104 sewage samples were collected from a treatment plant servicing approximately 500000 people and analysed for levels of methamphetamine, cocaine and cocaine metabolites. Careful examination of the kinetics of drug-flow profiles was then performed in order to identify trends or patterns of use within the community. RESULTS Results were validated against identical measurements of pharmaceutical reference compounds. Consumption profiles for cocaine and methamphetamine were found to differ in terms of frequency and timing of use. The majority of cocaine consumption occurs during the evening hours and 45% of consumption of this drug occurs in weekend periods. The flow of methamphetamine in the sewage system appears more evenly spread throughout the week. CONCLUSIONS This result is consistent with both an extended excretion half-life and a pattern of use that is more evenly balanced across all days of the week. Comprehensive investigation in to the scale and kinetics of drug flow in a sewage stream can therefore provide valuable information, not only in terms of the volume of drug consumed, but also in terms of identifying differing usage-patterns over daily and weekly time-scales.

Analysis of new classes of recreational drugs in sewage: Synthetic cannabinoids and amphetamine-like substances

The analysis of sewage for the residues of commonly used illicit drugs has successfully been applied as a suitable approach for estimating community illicit drug use. The drug market is increasingly dynamic with new substances continually being marketed for recreational purposes. In this study, ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to simultaneously and quantitatively detect the exogenous biomarkers of new classes of recreational drugs in sewage collected from three different Norwegian cities (Oslo, Bergen, Hamar). The samples were screened for the presence of khat (d-norpseudoephedrine and cathinone), mephedrone, pseudoephedrine, 7-aminoflunitrazepam, para-methoxyamphetamine (PMA), para-methoxy-N-methylamphetamine (PMMA) and a selection of urinary metabolites of synthetic cannabinoids collectively termed ´Spice´ (5-3-1-naphthoyl-1H-indol-1-yl-pentanoic acid (JWH 018 N-pentanoic acid), 1-5-hydroxypentyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 018 N-5-hydroxypentyl), 4-3-1-naphthoyl-1H-indol-1-yl-butanoic acid (JWH 073 N-butanoic acid), 1-4-hydroxybutyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 073 N-4-hydroxybutyl), 1-5-hydroxypentyl-1H-indol-3-yl4-methylnaphthalen-1-yl-methanone (JWH 122 N-5-hydroxypentyl), 1-5-fluoro-4-hydroxypentyl-1H-indol-3-ylnaphthalen-1-ylmethanone (AM2201 N-4-hydroxypentyl), and 1-5-hydroxypentyl-1H-indol-3-yl4-methoxyphenyl-methanone (RCS-4 N-5-hydroxypentyl)). Limits of detection were 1 ng/L for amphetamine like compounds and 5 ng/L for the metabolites of synthetic cannabinoids while the limits of quantification were 3 and 15 ng/L, respectively. Three of the fourteen selected biomarkers (cathine, pseudoephedrine and the synthetic cannabinoid metabolite JWH-018 N-5-hydroxypentyl) were detected in sewage, alongside the illicit drugs (and/or metabolites) typically found in sewage (cocaine, benzoylecognine, methamphetamine, MDMA, and THC-COOH). The khat biomarker d-norpseudoephedrine was detected in Oslo sewage at a mean concentration of 93 ng/L that represents a daily load of 54 mg/day/1000 inhabitants. Pseudoephedrine was present at mean concentrations of between 27 and 67 ng/L representing normalized daily loads of between 10 (Hamar) and 24 mg/day/1000 inhabitants (Bergen). The daily normalized loads of JWH-018 N-5-hydroxypentyl were between 49 (Oslo) and 62 mg/day/1000 inhabitants (Hamar). This study demonstrates for the first time that sewage biomarker analysis can be applied to evaluate not only the use the traditional illicit drugs (cocaine, cannabis and amphetamines), but also the use of certain new synthetic drugs.

What else can the analysis of sewage for urinary biomarkers reveal about communities

P health monitoring and evaluation is an essential part of all community health programs with the overall purpose of mapping the current level of health in a community and measuring the success of programs aimed at improving it. Public health agencies worldwide face the significant challenge as how to quickly and reliably evaluate the overall health of a population. One solution may simply lie beneath our feet. Recently an approach based on the analysis of specific urinary biomarkers in wastewater collated from defined populations has been used to determine the level of illicit drug use and even proposed as a method for drug testing athletes at the 2012 Olympic Games. This approach, which has attracted the attention of international drug monitoring agencies, such as the European Monitoring Centre for Drug and Drug Additction (EMCDDA; www.emcdda.europa.eu/wastewater-analysis), also has the potential to provide a means of rapidly (and quantitatively) assessing factors related to health and illness within a community. In its current form, the quantitative measurement of health specific biomarkers (i.e., drug biomarkers), in the collective sewage from populations, has been used to study the illicit drug use habits of numerous communities. The level of drug residues present in the sewage reflects the amount of a particular drug that has been consumed by the population served by the sewer network under investigation. The approach has been applied in Italy, Belgium, Spain, UK, Croatia, France, Ireland, Norway, USA, Canada, and Australia yielding promising results. More recently a similar approach has been used for legalized drugs, such as nicotine and alcohol that provide very specific data on their use in society. As some of the issues associated with obtaining representative sampling are resolved through collecting either high-resolution flow dependent samples or passive samples, depending on the level of resolution required, it may be possible to not only develop markers for other classes of drugs but also for other urinary biomarkers of human health (Table 1). As an example, excessive drinking is associated with major health problems so government agencies utilize measures such as increased taxes and restricted hours-of-sale in an attempt to reduce the overall volume of alcohol consumed. The quantitative measurement of specific biomarkers in sewage has been used to study the rates of alcohol consumption in defined populations, and could be used in the future to monitor the success of governmental consumption reduction measures or indeed the amount of bootleg alcohol entering the marketplace. When applied to specific urinary biomarkers of human health it has the potential to not only provide estimates of a broad number of lifestyle factors that influence health but also information about diet, health, disease and environment (Table 1). It is clearly possible that selected urinary biomarkers (e.g., those listed in Table 1) are suitable as wastewater biomarkers and will allow an assessment of the factors that influence population health and that the measurement of these biomarkers in sewage is a suitable approach for quantitatively evaluating factors that directly affect the health of communities. Quantitative analysis of ‘health biomarkers’ in sewage has the potential to provide evidencebased and real-time estimates of community/population health and may allow the monitoring of changes over time, for example in response to specific campaigns, identify new and changing trends as well as inter-community comparisons in combination with traditional epidemiological measurements. It is entirely possible that different communities will provide different patterns of wastewater biomarkers which will in turn relate to their health, diet, lifestyle and environment. We propose that the intercommunity/population comparison of a ‘sewage biomarker fingerprint’ will allow an assessment of the health related factors that differentiate those communities/populations. As with any emerging field of research there is a great deal of research that needs to be performed for this to be realized. For example, the need to evaluate the stability and specificity of selected biomarkers in sewage and the development of robust methods for their

The occurrence of second generation anticoagulant rodenticides in non-target raptor species in Norway.

Second generation anticoagulant rodenticides (SGARs) are commonly used for rodent pest control in Norway resulting in the potential exposure of non-target raptor species. In this study the occurrence of flocoumafen, difethialone, difenacoum, bromadiolone and brodifacoum was determined in the livers of five species of raptors found dead in Norway between 2009 and 2011. The SGARs brodifacoum, bromadiolone, difenacoum and flocoumafen were detected in golden eagle (Aquila chrysaetos) and eagle owl (Bubo bubo) livers at a total SGAR concentration of between 11 and 255 ng/g in approximately 70% of the golden eagles and 50% of the eagle owls examined in this study. In the absence of specific golden eagle and eagle owl toxicity thresholds for SGARs, a level of >100 ng/g was used as a potential lethal range, accepting that poisoning may occur below this level. Thirty percent (7/24) of the golden eagle and eagle owl livers contained total SGAR residue levels above this threshold. Further estimation of the potential mortality impact on the sampled raptor populations was not possible.