Spatial variations in the consumption of illicit stimulant drugs across Australia: A nationwide application of wastewater-based epidemiology.

Obtaining representative information on illicit drug use and patterns across a country remains difficult using surveys because of low response rates and response biases. A range of studies have used wastewater-based epidemiology (WBE) as a complementary approach to monitor community-wide illicit drug use. In Australia, no large-scale WBE studies have been conducted to date to reveal illicit drug use profiles in a national context. In this study, we performed the first Australia-wide WBE monitoring to examine spatial patterns in the use of three illicit stimulants (cocaine, as its human metabolite benzoylecgonine; methamphetamine; and 3,4-methylendioxymethamphetamine (MDMA)). A total of 112 daily composite wastewater samples were collected from 14 wastewater treatment plants across four states and two territories. These covered approximately 40% of the Australian population. We identified and quantified illicit drug residues using liquid chromatography coupled with tandem mass spectrometry. There were distinctive spatial patterns of illicit stimulant use in Australia. Multivariate analyses showed that consumption of cocaine and MDMA was higher in the large cities than in rural areas. Also, cocaine consumption differed significantly between different jurisdictions. Methamphetamine consumption was more similar between urban and rural locations. Only a few cities had elevated levels of use. Extrapolation of the WBE estimates suggested that the annual consumption was 3tonnes for cocaine and 9tonnes combined for methamphetamine and MDMA, which outweighed the annual seizure amount by 25 times and 45 times, respectively. These ratios imply the difficulty of detecting the trafficking of these stimulants in Australia, possibly more so for methamphetamine than cocaine. The obtained spatial pattern of use was compared with that in the most recent national household survey. Together both WBE and survey methods provide a more comprehensive evaluation of drug use that can assist governments in developing policies to reduce drug use and harm in the communities.

Surface-assisted laser desorption ionization mass spectrometry techniques for application in forensics.

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is an excellent analytical technique for the rapid and sensitive analysis of macromolecules (>700 Da), such as peptides, proteins, nucleic acids, and synthetic polymers. However, the detection of smaller organic molecules with masses below 700 Da using MALDI-MS is challenging due to the appearance of matrix adducts and matrix fragment peaks in the same spectral range. Recently, nanostructured substrates have been developed that facilitate matrix-free laser desorption ionization (LDI), contributing to an emerging analytical paradigm referred to as surface-assisted laser desorption ionization (SALDI) MS. Since SALDI enables the detection of small organic molecules, it is rapidly growing in popularity, including in the field of forensics. At the same time, SALDI also holds significant potential as a high throughput analytical tool in roadside, work place and athlete drug testing. In this review, we discuss recent advances in SALDI techniques such as desorption ionization on porous silicon (DIOS), nano-initiator mass spectrometry (NIMS) and nano assisted laser desorption ionization (NALDI™) and compare their strengths and weaknesses with particular focus on forensic applications. These include the detection of illicit drug molecules and their metabolites in biological matrices and small molecule detection from forensic samples including banknotes and fingerprints. Finally, the review highlights recent advances in mass spectrometry imaging (MSI) using SALDI techniques.

Forensic intelligence framework--Part I: Induction of a transversal model by comparing illicit drugs and false identity documents monitoring.

Forensic intelligence is a distinct dimension of forensic science. Forensic intelligence processes have mostly been developed to address either a specific type of trace or a specific problem. Even though these empirical developments have led to successes, they are trace-specific in nature and contribute to the generation of silos which hamper the establishment of a more general and transversal model. Forensic intelligence has shown some important perspectives but more general developments are required to address persistent challenges. This will ensure the progress of the discipline as well as its widespread implementation in the future. This paper demonstrates that the description of forensic intelligence processes, their architectures, and the methods for building them can, at a certain level, be abstracted from the type of traces considered. A comparative analysis is made between two forensic intelligence approaches developed independently in Australia and in Europe regarding the monitoring of apparently very different kind of problems: illicit drugs and false identity documents. An inductive effort is pursued to identify similarities and to outline a general model. Besides breaking barriers between apparently separate fields of study in forensic science and intelligence, this transversal model would assist in defining forensic intelligence, its role and place in policing, and in identifying its contributions and limitations. The model will facilitate the paradigm shift from the current case-by-case reactive attitude towards a proactive approach by serving as a guideline for the use of forensic case data in an intelligence-led perspective. A follow-up article will specifically address issues related to comparison processes, decision points and organisational issues regarding forensic intelligence (part II).

The use of organic and inorganic impurities found in MDMA police seizures in a drug intelligence perspective.

Traditional forensic drug profiling involves numerous analytical techniques, and the whole process is typically costly and may be time consuming. The aim of this study was to investigate the possibility of prioritising techniques utilised at the Australian Federal Police (AFP) for the chemical profiling of 3,4-methylenedioxymethylamphetamine (MDMA). The outcome would provide the AFP with the ability to obtain more timely and valuable results that could be used in an intelligence perspective. Correlation coefficients were used to obtain a similarity degree between a population of linked samples (within seizures) and a population of unlinked samples (between different seizures) and discrimination between the two populations was ultimately achieved. The results showed that gas chromatography-mass spectrometry (GC-MS) was well suited as a single technique to detect links between seizures and could be used in priority for operational intelligence purposes. Furthermore, the method was applied to seizures known or suspected (through their case information) to be linked to each other to assess the chemical similarity between samples. It was found that half of the seizures previously linked by the case number were also linked by the chemical profile. This procedure was also able to highlight links between cases that were previously unsuspected and retrospectively confirmed by circumstantial information. The findings are finally discussed in the broader forensic intelligence context, with a focus on how they could be successfully incorporated into investigations and in an intelligence-led policing perspective in order to understand trafficking markets.

Using quantitative wastewater analysis to measure daily usage of conventional and emerging illicit drugs at an annual music festival.

INTRODUCTION AND AIMS: Wastewater analysis provides a non-intrusive way of measuring drug use within a population. We used this approach to determine daily use of conventional illicit drugs [cannabis, cocaine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA)] and emerging illicit psychostimulants (benzylpiperazine, mephedrone and methylone) in two consecutive years (2010 and 2011) at an annual music festival. DESIGN AND METHODS: Daily composite wastewater samples, representative of the festival, were collected from the on-site wastewater treatment plant and analysed for drug metabolites. Data over 2 years were compared using Wilcoxon matched-pair test. Data from 2010 festival were compared with data collected at the same time from a nearby urban community using equivalent methods. RESULTS: Conventional illicit drugs were detected in all samples whereas emerging illicit psychostimulants were found only on specific days. The estimated per capita consumption of MDMA, cocaine and cannabis was similar between the two festival years. Statistically significant (P < 0.05; Z = -2.0-2.2) decreases were observed in use of methamphetamine and one emerging illicit psychostimulant (benzyl piperazine). Only consumption of MDMA was elevated at the festival compared with the nearby urban community. DISCUSSION AND CONCLUSIONS: Rates of substance use at this festival remained relatively consistent over two monitoring years. Compared with the urban community, drug use among festival goers was only elevated for MDMA, confirming its popularity in music settings. Our study demonstrated that wastewater analysis can objectively capture changes in substance use at a music setting without raising major ethical issues. It would potentially allow effective assessments of drug prevention strategies in such settings in the future.

Forensic applications of desorption electrospray ionisation mass spectrometry (DESI-MS).

Desorption electrospray ionisation mass spectrometry (DESI-MS) is an emerging analytical technique that enables in situ mass spectrometric analysis of specimens under ambient conditions. It has been successfully applied to a large range of forensically relevant materials. This review assesses and highlights forensic applications of DESI-MS including the analysis and detection of illicit drugs, explosives, chemical warfare agents, inks and documents, fingermarks, gunshot residues and drugs of abuse in urine and plasma specimens. The minimal specimen preparation required for analysis and the sensitivity of detection achieved offer great advantages, especially in the field of forensic science.

The use of forensic case data in intelligence-led policing: the example of drug profiling.

To date, forensic science has predominantly focused on generating evidence for judicial proceedings. While many recognise its broader and important contribution to the initial stages of the forensic process, resources do not seem to be employed efficiently. It is often discovered retrospectively that necessary information was previously available in a database or within existing files. Such information could have been proactively used in order to solve a particular case, a number of linked cases or better understand the criminal activity as a whole. This article reviews this broader contribution of forensic science, with a particular emphasis on drug intelligence at the Australian Federal Police (AFP) in Australia. Using the AFP as a model organisation, an overview of the current situation and the contribution of physical and chemical profiling are first discussed. The situation in Europe, and in particular in Switzerland, is also presented. It is argued that a change of attitude towards a more intelligence-led perspective is required in forensic science in general, and in drug profiling in particular.

Profiles of illicit drug use during annual key holiday and control periods in Australia: wastewater analysis in an urban, a semi-rural and a vacation area.

AIMS: To examine changes in illicit drug consumption between peak holiday season (23 December-3 January) in Australia and a control period two months later in a coastal urban area, an inland semi-rural area and an island populated predominantly by vacationers during holidays. DESIGN: Analysis of representative daily composite wastewater samples collected from the inlet of the major wastewater treatment plant in each area. SETTING: Three wastewater treatment plants. PARTICIPANTS: Wastewater treatment plants serviced approximately 350, 000 persons in the urban area, 120,000 in the semi-rural area and 1100-2400 on the island. MEASUREMENTS: Drug residues were analysed using liquid chromatography coupled to a tandem mass spectrometer. Per capita drug consumption was estimated. Changes in drug use were quantified using Hedges' g. FINDINGS: During the holidays, cannabis consumption in the semi-rural area declined (g = -2.8) as did methamphetamine (-0.8), whereas cocaine (+1.5) and ecstasy (+1.6) use increased. In the urban area, consumption of all drugs increased during holidays (cannabis +1.6, cocaine +1.2, ecstasy +0.8 and methamphetamine +0.3). In the vacation area, methamphetamine (+0.7), ecstasy (+0.7) and cocaine (+1.1) use increased, but cannabis (-0.5) use decreased during holiday periods. CONCLUSIONS: While the peak holiday season in Australia is perceived as a period of increased drug use, this is not uniform across all drugs and areas. Substantial declines in drug use in the semi-rural area contrasted with substantial increases in urban and vacation areas. Per capita drug consumption in the vacation area was equivalent to that in the urban area, implying that these locations merit particular attention for drug use monitoring and harm minimisation measures.

An analysis of ethical issues in using wastewater analysis to monitor illicit drug use.

AIMS: To discuss ethical issues that may arise in using WWA to monitor illicit drug use in the general population and in entertainment precincts, prisons, schools and work-places. METHOD: Review current applications of WWA and identify ethical and social issues that may be raised with current and projected future uses of this method. RESULTS: Wastewater analysis (WWA) of drug residues is a promising method of monitoring illicit drug use that may overcome some limitations of other monitoring methods. When used for monitoring purposes in large populations, WWA does not raise major ethical concerns because individuals are not identified and the prospects of harming residents of catchment areas are remote. When WWA is used in smaller catchment areas (entertainment venues, prisons, schools or work-places) their results could, possibly, indirectly affect the occupants adversely. Researchers will need to take care in reporting their results to reduce media misreporting. Fears about possible use of WWA for mass individual surveillance by drug law enforcement officials are unlikely to be realized, but will need to be addressed because they may affect public support adversely for this type of research. CONCLUSIONS: Using wastewater analysis to monitor illicit drug use in large populations does not raise major ethical concerns, but researchers need to minimize possible adverse consequences in studying smaller populations, such as workers, prisoners and students.

Styryl dye coated metal oxide powders for the detection of latent fingermarks on non-porous surfaces.

Conventional fingermark powders rely on contrast induced by absorption/reflection (e.g. black powder) or luminescence in the visible region (e.g. Blitz Green(®)). In most cases, these powders provide sufficient contrast; however, in some circumstances surface characteristics can interfere with the visualisation of powdered fingermarks. Visualisation in the near infra-red (NIR) region, however, has been shown to eliminate interferences commonly encountered in the visible region. In this study, a mixture of rhodamine 6G and the NIR laser dye styryl 11 (STaR 11) was coated onto an aluminium oxide nanopowder and then mixed with silver magnetic powder to develop and visualise fingermarks in the NIR. When compared to Blitz Green(®), it was determined that the STaR 11 magnetic powder was better suited for marks deposited on textured surfaces and for older marks, whereas Blitz Green(®) performed better on smooth glossy surfaces. The ability of the STaR 11 mixed dye formulation to be visualised in both the visible and NIR regions also provides a significant advantage over conventional luminescent fingermark powders.

Screening of gunshot residues using desorption electrospray ionisation-mass spectrometry (DESI-MS).

Several studies have indicated that there are potential environmental sources of particles resembling inorganic primer found in gunshot residues (GSR); as a consequence examiners are reluctant to unambiguously assign the origin of inorganic particles. If organic gunshot residues (OGSR) were found in combination with inorganic particles, the possibility of environmental sources could be potentially eliminated, thereby significantly enhancing the strength of the evidence. Methods have been previously described whereby GSR specimens can be analysed for the presence of OGSR or inorganic GRS (IGSR). However, no methods have been reported that allow the analysis of both OGSR and IGSR on the same specimen. Described in this article is a direct method using desorption electrospray ionisation-mass spectrometry (DESI-MS) for the detection of methyl centralite (MC), ethyl centralite (EC) and diphenylamine (DPA) on adhesive tape GSR stubs typically used for scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) analysis. The optimisation of numerous parameters was conducted using an experimental design. The results indicate that direct analysis of these organic components of GSR is possible although some limitations were also identified. This initial investigation has also indicated that subjecting stubs to DESI analysis does not interfere with subsequent SEM-EDX analysis of primer residues; therefore the technique described herein allows a comprehensive examination of GSR that would be highly probative in the event that both OGSR and IGSR are detected in the same specimen.

Use of styryl 11 and STaR 11 for the luminescence enhancement of cyanoacrylate-developed fingermarks in the visible and near-infrared regions.

In current casework, most post-cyanoacrylate stains rely on luminescence emission in the visible region (400-700 nm). While traditional stains such as rhodamine 6G work well under most circumstances, some surfaces may generate background luminescence under the same conditions. Detection in the near-infrared region (NIR > 700 nm) has shown to be effective in minimizing the interferences from such surfaces. The laser dye styryl 11 generated strongly luminescent fingermarks when applied after cyanoacrylate fuming on all surfaces tested. When compared to rhodamine 6G, the dye was superior only when viewed in the NIR. Styryl 11 was subsequently combined with rhodamine 6G, and the mixed stain formulation (named StaR 11 by the authors) induced stronger luminescence compared with styryl 11 alone with an ability to visualize in both the visible and NIR regions. Reliable and consistent results were obtained when using either styryl 11 alone or the STaR 11 mixture. The enhancement achieved did not otherwise vary depending on the source of the fingermark secretions. With visualization possible in both the visible and NIR regions, the styryl 11/rhodamine 6G mixture showed significant potential as a post-cyanoacrylate stain.

Refining the estimation of illicit drug consumptions from wastewater analysis: co-analysis of prescription pharmaceuticals and uncertainty assessment.

Wastewater analysis is a promising monitoring tool to estimate illicit drug consumption at the community level. The advantage of this technique over traditional surveys and other surveillance methods has been emphasized in recent studies. However, there are methodological challenges that can affect reliability. The objectives of this study were to systematically reduce and assess uncertainties associated with sampling (through a stringent optimization of the sampling method) and the back calculation of per capita drug consumption (through a refined estimation of the number of people actively contributing to the wastewater in a given period). We applied continuous flow-proportional sampling to ensure the collection of representative raw wastewater samples. Residues of illicit drugs, opioids, prescription pharmaceuticals and one artificial sweetener were analyzed by liquid chromatography coupled with tandem mass spectrometry. A parameter estimating the number of people actively contributing to wastewater over a given period was calculated from the measured loads of prescription pharmaceuticals, their annual consumption and relative excretion data. For the calculation of substance loads in sewage, uncertainties were propagated considering five individual components: sampling, chemical analysis, flow measurements, excretion rates and the number of people contributing to the wastewater. The daily consumption per 1000 inhabitants was estimated to be almost 1000 mg for cannabis and several hundred mg for cocaine, methamphetamine and ecstasy. With the best sampling practice and current chemical analysis, we calculated the remaining uncertainty to be in the range of 20-30% (relative standard deviation, RSD) for the estimation of consumed drug masses in the catchment; RSDs for the per capita consumption were lower (14-24%), as one of the biggest uncertainty components (i.e. error in flow measurements) cancels out in the proposed method for the estimation of the number of people contributing to the daily wastewater volume. In this study, we provide methodological improvements that substantially enhance the reliability of the estimation method--a prerequisite for the application of this technique to meaningfully assess changes in drug consumption and the success of drug intervention strategies in future studies.

Combined immunocapture and laser desorption/ionization mass spectrometry on porous silicon.

There is considerable interest in the highly parallelized mass spectrometry analysis of complex sample mixtures without any time-consuming prepurification. Porous silicon-based laser desorption/ionization mass spectrometry (pSi LDI-MS) is enabling technology for such analysis. Previous studies have focused on pSi surface functionalization to enhance sensitivity of detection and engineer surfaces for sample capture and enrichment in LDI-MS analysis. In this report, we build on this work by showing that surface functionalization of thin pSi films can be extended to the covalent immobilization of antibodies, producing a porous immunoaffinity surface. We demonstrate highly selective mass spectrometric detection of illicit drugs (benzodiazepines) on pSi films displaying antibenzodiazepine antibodies covalently immobilized via isocyanate chemistry. The effects of antibody immobilization conditions, antibody concentration, and surface blocking on LDI-MS performance and selectivity were studied. X-ray photoelectron spectroscopy (XPS) was instrumental in characterizing surface chemistry and optimizing LDI-MS performance. Overall, our approach is suitable for rapid and sensitive confirmatory analysis in forensic toxicology requiring only minimal sample volume and may be applied to other areas requiring small molecular analysis such as metabolomics and pharmacology.

Rapid drug detection in oral samples by porous silicon assisted laser desorption/ionization mass spectrometry.

The demand for analysis of oral fluid for illicit drugs has arisen with the increased adoption of roadside testing, particularly in countries where changes in legislation allow random roadside testing of drivers for the presence of a palette of illicit drugs such as methamphetamine (MA), 3,4-methylenedioxymethamphetamine (MDMA) and Delta9-tetrahydrocannabinol (THC). Oral samples are currently tested for such drugs at the roadside using an immunoassay-based commercial test kit. Positive roadside tests are sent for confirmatory laboratory analysis, traditionally by means of gas chromatography/mass spectrometry (GC/MS). We present here an alternative rapid analysis technique, porous silicon assisted laser desorption/ionization time-of-flight mass spectrometry (pSi LDI-MS), for the high-throughput analysis of oral fluids. This technique alleviates the need for sample derivatization, requires only sub-microliter sample volumes and allows fast analysis (of the order of seconds). In this study, the application of the technique is demonstrated with real samples from actual roadside testing. The analysis of oral samples resulted in detection of MA and MDMA with no extraction and analysis of THC after ethyl acetate extraction. We propose that, subject to miniaturization of a suitable mass spectrometer, this technique is well suited to underpin the deployment of oral fluid testing in the clinic, workplace and on the roadside.

Forensic analysis of condom and personal lubricants by capillary electrophoresis.

Condoms may offer sexual assailants a simple and relatively effective means by which they may remove and dispose of the biological evidence of their contact with the victim. Without this valuable probative evidence, the investigator may need to turn to such alternative forms of physical evidence, such as the residue remaining from the condom lubricant, and any other additional lubricating substances, which may have been used by the assailant. In this study, 68 different condom and personal lubricant samples were analysed using micellar electrokinetic capillary chromatography (MEKC) with ultraviolet absorbance detection. The electropherograms were processed by principal component analysis (PCA) and classified with linear discriminant analysis. This classified 233 samples out of 263 into their correct sample group. An alternative method of classification in which the sample electropherograms were converted to vectors and a correlation was determined by the cosine of the angles between these vectors. This correctly classified 172 samples of the 296. Results indicate that the combination of CE with chemometric data treatment offers the potential of being a rapid and efficient means by which condom and personal lubricant samples may be differentiated. Although this analytical method at present lacks the sensitivity required for sexual assault swab analysis, it is hoped that future developments in instrumentation detection capabilities will permit its use one day as a routine casework tool.

A method for the longitudinal sectioning of single hair samples.

A simple technique is presented for the longitudinal sectioning of hair samples without the need for any embedding medium. The technique applies to single hair samples that are placed in a groove of a particular depth and can either be scraped or cut to the desired level. Planar sections are obtained that reveal the internal structure of the sample and are ideal for the application of surface analytical techniques for the study of internal transverse and longitudinal distributions of xenobiotics.

The contribution of the metabolite p-hydroxyamphetamine to the central actions of p-methoxyamphetamine.

RATIONALE: Para-methoxyamphetamine (PMA) is a substituted amphetamine that has been responsible for a number of fatalities in Australia and North America. Previous investigators have shown that p-hydroxyamphetamine (PHA), the primary metabolite of PMA, has effects on central neurotransmitter kinetics in vitro that are similar to those of the parent compound. In order to understand the role of PHA, it is necessary to determine both the in vivo actions and the concentrations achieved relative to those of PMA. OBJECTIVES: The effects of PHA and PMA on 5-hydroxytryptamine (5HT) and dopamine kinetics in brain were determined and the concentrations of each compound measured in blood and brain. METHODS: Animals were housed at 20-22C on a standard 12/12-h light/dark cycle. High speed chronoamperometry was used to compare the ability of PMA and PHA to alter 5HT and dopamine kinetics in the rat striatum in vivo. Concentrations of PHA and PMA in blood, whole brain and striatum were determined following a dose of PMA (10 mg/kg, IP.) using HPLC with fluorescence detection. RESULTS: PHA was more effective than PMA at evoking neurotransmitter release and inhibiting the uptake of dopamine. However, both compounds were approximately equipotent 5HT uptake inhibitors. PMA and PHA concentrations in whole brain and striatum peaked within 30 min of the administered dose, whereas blood concentrations of both compounds peaked 1 h after the dose. PHA concentrations in both blood and brain were consistently much lower than PMA concentrations. CONCLUSIONS: These data indicate that although PHA is more effective than PMA at altering 5HT and dopamine kinetics in vivo, it is unlikely to achieve sufficient brain concentrations to contribute to the central effects of PMA.