Seized Drug Samples Research Articles

Emergence of semi-synthetic cannabinoids in cannabis products seized in Eastern Denmark over a 6-year period.

Semi-synthetic cannabinoids (SSCs) are derivatives of phytocannabinoids with slight chemical modifications. SSCs have appeared as legal alternatives to tetrahydrocannabinol (Δ9-THC) in recent years. This study investigates the prevalence of SSCs in seized drug samples from Danish police and custom authorities seized in Eastern Denmark in the period 2018-2023. Screening data obtained by gas chromatography-mass spectrometry (GC-MS) were reprocessed to enable detection of SSCs. Seized drug samples were categorized into six types of formulations. Δ8-THC was the first SSC observed and appeared in 2019 followed by hexahydrocannabinol (HHC), tetrahydrocannabidiol (H4-CBD), hexahydrocannabinol acetate (HHC-O-Acetate), hexahydrocannabiphorol (HHCP) and tetrahydrocannabiphorol (Δ9-THCP). Only one sample positive for SSCs was observed before the third quarter of 2021, with positive samples increasing from third quarter of 2022. Over the study period, a total of 15% (n = 216) of seized cannabis products were positive for SSCs. HHC was the most frequently identified SSC and found in 10% (n = 137) of samples, followed by H4-CBD at 4% (n = 53), Δ8-THC at 3% (n = 44), and HHC-O-Acetate, HHCP, and THCP each at 1% (n = 10-20). SSCs appeared in 56% of E-cigarette products, 20% of hashish, 17% of concentrates, 10% of edibles, and 10% of plant materials. In conclusion, SSCs represent a new type of cannabinoids with a rapidly growing popularity and with specific compounds dominating at different periods. Some of the observed trends were likely influenced by the scheduling of HHC in May of 2023 in Denmark.

Screen-printed electrode-based sensor for rapid ketamine determination: optimization and on-site application for seized drugs analysis

This study investigates the electrochemical behavior of ketamine using an in-lab fabricated screen-printed electrode system and explores its potential application in quantitative analysis. Cyclic voltammetry and differential pulse voltammetry (DPV) experiments were employed to characterize the oxidation behavior of ketamine. Systematic optimization of DPV parameters, including pulse amplitude, pulse width, potential step, potential, and time accumulation for analyte preconcentration resulted in the selection of optimal conditions for quantitative analysis. The developed DPV method exhibited excellent linearity (R2 = 0.996) over the concentration range of 50–500 µM, with a limit of detection of 15 µM and a limit of quantification of 50 µM. Authentic samples analysis demonstrated the utility of the proposed sensor for quantitative analysis of ketamine in pharmaceutical products and seized drug samples. Overall, the developed sensor offers a promising tool for the rapid and accurate analysis of ketamine in various samples with potential applications in on-site forensic analysis.Graphical abstract

Enantiomeric profile of amphetamines in seized drug samples and in blood of impaired drivers in Iceland: The rise of (R)-methamphetamine?

Amphetamine (AMP) and methamphetamine (METH) use is increasing globally. Illegal AMP is generally a racemic mixture, whereas AMP-containing attention-deficit hyperactivity disorder drugs prescribed in Iceland consist of S-AMP. AMP is also a main metabolite of interest after METH intake. Distinguishing between legal and illegal AMP intake is vital in forensic toxicology. A chiral UPLC-MS-MS method was used to determine the enantiomeric profile of AMP and METH in circulation in Iceland by analysing blood samples from drivers suspected of driving under the influence of drugs (DUID) and seized drug samples from 2021 and 2022. All seized AMP samples (n = 48) were racemic, whereas all but one seized METH sample (n = 26) were enantiopure. Surprisingly, a large portion of the enantiopure METH samples was R-METH. DUID blood samples positive for AMP (n = 564) had a median blood concentration of 180 ng/mL (range 20-2770 ng/mL) and a median enantiomeric fraction (EFR) of 0.54 (range 0-0.73), whereas samples positive for METH (n = 236) had a median blood concentration of 185 ng/mL (range 20-2300 ng/mL) and a median EFR of 0.23 (range 0-1). The findings of this study show a significantly lower blood concentration in drivers with only S-AMP detected compared with when the R-isomer is also detected. No significant difference in blood concentration was detected between the sample groups containing S-METH, R-METH or both enantiomers. The occurrence of R-METH in both seized drug samples and DUID cases indicates a change in drug supply and a need for better scientific knowledge on R-METH abuse.

Interlaboratory validation and one-year retrospective review of hemp and marijuana decision-point assays

After the passage of the Agriculture Improvement Act of 2018 (Farm Bill) that defined hemp as Cannabis sativa L. if it contained ≤0.3 % Δ9-THC on a dry weight basis, forensic laboratories were faced with the challenge of distinguishing marijuana from hemp in seized drug samples. This study reports the results of an interlaboratory validation of a previously developed qualitative decision-point assay for the differentiation of illegal marijuana from legal hemp, utilizing gas chromatography-mass spectrometry (GC–MS) with a 1 % Δ9-THC threshold. The method was validated in terms of selectivity, linearity, carryover, precision, accuracy, extract stability, dilution integrity, and measurement uncertainty. Other elements such as decarboxylation rate and potential for CBD interference were also evaluated. Specificity and positive predictive value were both 100 % using known cannabis reference materials. A total of 8 false negative results were observed among the 280 analyses, resulting in an overall assay sensitivity of 94 % and a negative predictive value of 95 %. The small number of false negative results near the 1 % threshold were attributed to decarboxylation inefficiency in some laboratories. While qualitative in nature, the measurement uncertainty of the assay at the 0.3 % Δ9-THC cutoff (legal threshold) using a 95.45 % confidence interval (k = 2) ranged from 12.2 % to 21.8 % among laboratories (equivalent to 0.3 % ± 0.04 % to 0.07 % Δ9-THC by weight). A one-year retrospective study across three agencies and fifteen laboratories was undertaken. Among the three agencies participating in the original study, 93–96 % of all suspected cannabis seizures yielded results above the administrative threshold of 1 % Δ9-THC (n = 3,288). The interlaboratory method development highlighted differences in performance between sites, ultimately improving the overall robustness of the method. Observations highlight the need for site-specific validation and vigilance. While multi-agency collaborations and interlaboratory validations are valuable, they do not replace the necessity for full, independent, and rigorous validation.

An LC-MS/MS method for the determination of W18 in urine samples

IntroductionSynthetic drugs pose one of the most significant drug problems worldwide. In this category, W18 emerges as a potent drug of abuse chemically related to fentanyl. W18 has an analgesic potency 10,000 times greater than morphine. Recent in-vitro studies reported no activity of W18 towards opioid receptors. However, its presence in seized drug samples indicates its use as a precursor in fentanyl synthesis. This emphasizes the need to develop methods for its detection in developing countries dealing with emerging new drugs.ObjectivesTo develop an analytical method for the determination of W18 in urine samples.MethodsStandards with W18 concentrations ranging from 5-500 ng/ml were prepared in negative urine along with deuterated internal standard. The samples were diluted with methanol, centrifuged and the supernatant was subjected to Liquid chromatography-tandem mass spectrometry (LC-MS-MS) with time of flight (QTOF) analysis. For chromatographic separation, a C18 column with 50 degrees oven temperature was used. The mobile phase consists of formic acid, water, and acetonitrile. The TOF MS was operated in positive ion mode and multiple reaction monitoring was used for quantification.ResultsThe retention time of W18 was obtained at 9.57 minutes. The parent ion with molecular weight 422.1 along with precursor ions Q1-273, Q2-111.0, Q3-150.0 g/mol were measured. The area of the standards ranges from 1 to 9.0 log 5 with R square of 0.99. The limit of detection (LOD) and quantitation were 5 and 20 ng/ml respectively. The recovery of W18 was estimated to be 96% from the from spiked urine standards.ConclusionsThe developed method is able to detect W18 presence in urine samples. This method has the potential to be used in clinical and research studies.Disclosure of InterestNone Declared

Developing Aptamer-Based Colorimetric Opioid Tests.

Opioids collectively cause over 80,000 deaths in the United States annually. The ability to rapidly identify these compounds in seized drug samples on-site will be essential for curtailing trafficking and distribution. Chemical reagent-based tests are fast and simple but also notorious for giving false results due to poor specificity, whereas portable Raman spectrometers have excellent selectivity but often face interference challenges with impure drug samples. In this work, we develop on-site sensors for morphine and structurally related opioid compounds based on in vitro-selected oligonucleotide affinity reagents known as aptamers. We employ a parallel-and-serial selection strategy to isolate aptamers that recognize heroin, morphine, codeine, hydrocodone, and hydromorphone, along with a toggle-selection approach to isolate aptamers that bind oxycodone and oxymorphone. We then utilize a new high-throughput sequencing-based approach to examine aptamer growth patterns over the course of selection and a high-throughput exonuclease-based screening assay to identify optimal aptamer candidates. Finally, we use two high-performance aptamers with KD of ∼1 μM to develop colorimetric dye-displacement assays that can specifically detect opioids like heroin and oxycodone at concentrations as low as 0.5 μM with a linear range of 0-16 μM. Importantly, our assays can detect opioids in complex chemical matrices, including pharmaceutical tablets and drug mixtures; in contrast, the conventional Marquis test completely fails in this context. These aptamer-based colorimetric assays enable the naked-eye identification of specific opioids within seconds and will play an important role in combatting opioid abuse.

N-Isopropylbenzylamine-induced conditioned place preference, sensitization behaviour and self-administration in rodents.

N-Isopropylbenzylamine (N-ipb), a chain isomer of methamphetamine (METH) with similar physical properties, has been used as a substitute for METH in seized drug samples. However, the abuse potential of N-ipb remains unclear. Therefore, this study aimed to evaluate the abuse potential of N-ipb in comparison to METH, by using conditioned place preference (CPP), locomotor sensitization and intravenous self-administration tests. The results showed that N-ipb at a dose of 3 mg·kg-1 significantly induced CPP in mice, which was comparable to the effect of METH at 1mg·kg-1 . Either acute or repeated N-ipb injections (1 or 3mg·kg-1 ) failed to raise the locomotor activity. However, acute treatment with 10mg·kg-1 N-ipb elevated the locomotor activity compared with saline, while chronic injection of 10mg·kg-1 N-ipb induced a delayed and attenuated sensitization compared with 1mg·kg-1 METH. Rats could acquire N-ipb self-administration at a dose of 1mg·kg-1 ·infusion-1 , and a typical inverted U-shaped dose-response curve was obtained for N-ipb. The mean dose of N-ipb that maintained the maximum response was greater than that of METH, indicating that N-ipb is less potent for reinforcement than METH. In the economic behavioural analysis, comparison of essential values derived from the demand elasticity revealed that N-ipb is less efficacy as a reinforcer than METH. The present data demonstrate that N-ipb functions as a reinforcer and has a potential for abuse. However, the potency of psychomotor stimulation and the reinforcing effectiveness of N-ipb are lower than those of METH.

Desalkylgidazepam Blood Concentrations in 63 Forensic Investigation Cases.

Desalkylgidazepam, also known as bromonordiazepam, is the latest designer benzodiazepine to appear in postmortem blood samples in British Columbia. Our laboratory was first alerted to the presence of desalkylgidazepam in seized drug samples in May 2022 and the analyte was added to an in-house library shortly thereafter. Previously acquired spectra from routine death investigation cases were reprocessed using the updated library with the first presumptive identification of desalkylgidazepam occurring in a sample received in April 2022. A standard addition method for the quantitation of desalkylgidazepam in blood samples (from femoral, iliac, jugular and subclavian veins) was validated and consequently used to confirm presence and concentrations of the drug in 63 cases, with an average concentration of 42.2 ± 44.0 ng/mL (median concentration: 24.5 ng/mL; range: 3.7 - 220.6 ng/mL). Similar to detections of other novel benzodiazepines, co-occurrence of desalkylgidazepam with opioids and/or stimulants was common. To our knowledge, this paper is the first to report desalkylgidazepam concentrations in postmortem blood samples.

Evaluation of fentanyl immunoassay test strips for rapid in-situ detection of fentanyl and fentanyl analogs in seized samples and alternative matrices.

Ion mobility spectrometry is used for the rapid detection of drugs at points of security but are unable to differentiate some drugs leading to the instrument alarming for a drug not present in the sample. This can be particularly problematic for samples that alarm for fentanyl. In this study, fentanyl immunoassay strips were evaluated for use as a secondary test for fentanyl, including for the testing of alternative matrices, such as powders, e-liquids, and infused papers and textiles. The limit of detection of fentanyl immunoassay strips was examined along with their selectivity to 18 fentanyl analogsand 72 other drugs and cutting agents. The effectiveness of the test strips at the detection of fentanyl in the presence of other drugs was examined by testing a series of concentrations of fentanyl in solution in combination with other drugs. The testing of alternative matrices was explored with laboratory prepared samples through sampling with cotton buds and extraction in water. The fentanyl immunoassay strips detected fentanyl at concentrations of 45 ng/mL and reacted with 16 of 18 tested fentanyl analogs with carfentanil and norfentanyl being the only analogs to not react. There was no reactivity with other drugs or cutting agents. The effectiveness of the fentanyl test strips was not reduced when fentanyl was mixed with other drugs. Fentanyl was successfully detected with high sensitivity in all alternative matrices. The fentanyl immunoassay strips were found to be an effective secondary test for fentanyl and at least 16 fentanyl analogs in seized drug samples, including when mixed with other drugs. The effectiveness of the sampling methods for alternative matrices should be further evaluated using fentanyl and fentanyl analog casework samples. The use of this method by law enforcement and other agencies should be examined to assess its effectiveness and ease of use in operational settings.

GC-IR technique a rapid tool for detection of Tramadol in Forensic Confiscated Drugs and Biological Fluids

Tramadol is placed under Drug Controlled Substance Act, in India since 2018, owing to its extensive abuse as a narcotic substance recently. Subsequently, there is increased illegal trafficking across our nation for the drug tramadol. Forensic Science Laboratory, Delhi receives confiscated drugs and toxicological samples for chemical analysis to evaluate their precise composition enabling regulations from law enforcement agencies. Although considered a safe drug for its anti-inflammatory properties when used under medical guidance nevertheless, unintentional fatal tramadol intoxication due to its action on opioid receptors has been reported in several autopsy cases. Contraband tramadol is presently being examined using Gas Chromatography-Mass Spectrometer (GC-MS) and Fourier-transform infrared spectroscopy (FTIR) instrumental techniques in routine forensic examination. In the proposed study, an advanced sensitive technique Gas Chromatography Infrared spectroscopy (GC-IR) was explored as an alternative tool for rapid identification and confirmation. Standard Tramadol was used for method establishment and the method was subsequently applied to seized drug samples and Biological Fluids (blood, and urine samples). Findings of the study indicate that this method can be successfully applied to seized drugs and biological toxicological samples in the forensic examination for the drug Tramadol which was successfully detected by GC-IR. This new rapid analytical method will prove beneficial for the forensic community during routine examination of forensic samples with confirmation.

Development and validation of a rapid GC–MS method for seized drug screening

Controlled substances are one of the most commonly analyzed types of evidence in forensic laboratories. Persistent, high levels of case submissions in recent years have created a need for rapid screening techniques to decrease case backlogs and expedite confirmatory analyses. The most utilized technique for the analysis of seized drugs is gas chromatography mass spectrometry (GC-MS). While this approach typically takes tens of minutes, advances in the technique have enabled approximately 1 min run times with a short column and a rapid (10’s °C/s) temperature program. These advances were used in this work to develop and validate a rapid GC-MS method for screening seized drug samples. Method validation was completed to determine retention time repeatability and reproducibility, limits of detection, identification accuracy, selectivity, and carryover. Validation results showed the utility of this method for accurate and precise qualitative analyses for the screening of seized drugs across seven different drug classes. The optimized method was then applied to 15 real and mock case samples from the Maryland State Police Forensic Sciences Division. Controlled substances, cutting agents, and other diluents were identified from these samples using acceptance criteria of retention time and mass spectral comparisons currently used in casework. Using rapid GC-MS methods in casework could enable faster and more reliable presumptive testing results for the analysis of seized drug exhibits.

Forensic implication of seized drug imitating methamphetamine with mileage in crime

BackgroundThe increase in drug abuse a major global issue. Clandestine laboratories along with illegal drug trafficking and trade increase the menace. Increased confiscation of illicit drugs not only indicates the manifold rise in drug abuse in Delhi and its neighboring region but also signifies an escalation and proliferation in other drug-related criminal activities. Investigating agencies around the world are working hard to combat and eradicate this problem. The samples seized by these agencies are subsequently sent for forensic examination to rule out the presence of any illicit drugs. Few such seized crystalline samples such as “methamphetamine” were submitted for examination with a query for the identification of any scheduled substance. The analysis outline followed was physical, chemical, Fourier transform infrared spectrometer analysis for detection, identification, and confirmation of seized drug samples. The purpose of a detailed investigation regarding the identification of the crystalline sample was also to understand the criminal activities involved in the consistent seizures and the forensics behind it.ResultsThe comprehensive examination identified the sample as an inorganic salt of sodium thiosulfate with the absence of contraband methamphetamine. The study on salt-imitating methamphetamine underlines its diverse uses as in adulteration, trafficking to camouflage narcotics drug and its involvement in numerous other illegal purposes which on the other hand has a very crucial impact on society and its well-being.ConclusionsThe novel findings will update the investigating agencies and other experts regarding the importance of the detection of sodium thiosulfate salt in seized samples and will well establish the foul play behind large seizures to justify the role of forensic science. This unique finding deals with a very sensitive issue that has immense social impact and needs exceptional thoughtfulness.

Comprehensive profiling of controlled substances using gas chromatography coupled with time-of-flight mass spectrometry

Background: Drug profiling is performed to prove the presence of any controlled substances and to compare sample purity, to determine whether two samples are from the same origin, as well as to assess the expected monetary value of the drug samples. However, conventional analyses by gas chromatography – mass spectrometry (GC-MS) often lacks the sensitivity required to provide a full chemical fingerprint of the trace contaminants and cutting agents. Here, we apply gas chromatography coupled with time-of-flight mass spectrometry (GC-TOF MS) to identify trace differences between seized drug samples. Material and Methods: Six seized samples of suspected drugs of abuse, were prepared by dissolving of each drug powder in methanol, prior to analysis by GC–TOF MS. Resulting chromatograms were integrated, identified and compared using novel comparative analysis tools to determine trends and differences between the seized samples. Result: All six seized samples were found to contain diacetylmorphine (heroin), however the trace cutting agents and contaminants present varied significantly. For example, the presence of the common cutting agents, caffeine and acetaminophen (or paracetamol) was only noted for some of the samples, while others contained trace impurities, such as meconin and 6-acetylcodeine, which can be useful in linking samples to the same origin. Conclusion: Comprehensive profiling of seized drug samples was obtained using GC–TOF MS to identify not only the controlled substances, but also the various cutting agents and trace impurities present.

A survey of Eutylone Prevalence in New Zealand during 2020 and 2021

This study provides an insight into the prevalence of eutylone in New Zealand by analysis of the post-mortem forensic toxicology cases over the period 2020 and 2021, as well as analysis of suspected drug items collected from three music festivals between December 2020 and February 2021. Eutylone, also known as bk-EBDB or n-ethylbutylone, is a synthetic cathinone that has no known therapeutic use. In New Zealand it is sold recreationally as an alternative to methylendioxymethamphetamine (MDMA) and has increasingly been found as an adulterant or substitution substance in MDMA samples. Eutylone is known to cause harm and has been associated with fatalities [World Health Organization. “WHO Expert Committee on Drug Dependence: forty-fourth report.” (2022)]. In many cases, users are unaware that the eutylone is present believing they are taking MDMA. ESR undertakes post-mortem forensic toxicology casework for the whole of New Zealand. We investigated the post-mortem cases received during 2020 and 2021 for eutylone and investigated trends. In addition to the post-mortem forensic toxicology cases, ESR analysed 656 drug samples from three music festivals held between December 2020 and February 2021 for the presence of drugs including eutylone. The post-mortem blood samples were analysed using liquid chromatography with time of flight mass spectrometry (LC-TOFMS) for eutylone. Eutylone was detected in three cases during 2020 and in twelve (12) cases during 2021. Of the 12 cases from 2021, ten decedents were male, nine cases had a known cause of death (e.g. hanging), two cases were at or just after a music festival and two cases appeared to be drug overdoses where multiple drugs were involved. Eutylone was the only drug detected in three of the cases (all with a known cause of death). When other drugs were involved the most common were methamphetamine ( n = 4), MDMA ( n = 3), alcohol ( n = 3) and cannabis ( n = 3). For the 656 samples analysed from the three music festivals MDMA and eutylone were detected in 33% and 32% of the samples, respectively. This can be contrasted with a previous survey from December 2018 to March 2019 that found MDMA in 64% of samples and eutylone detected in less than 1% of the samples. If a mixture was present in a sample, it was common for it to contain both eutylone and MDMA. There was a significant increase in eutylone detections both in blood samples from post-mortem forensic toxicology casework and in seized drug samples over the time period studied, with associated increases in the harm, including deaths, associated with the drug use. It has been postulated that this trend was influenced by factors such as the limited supply of MDMA in New Zealand due to law enforcement activity including a large MDMA seizure prior to the survey period for the music festivals, COVID-19 restrictions and the use of eutylone as an adulterant or replacement drug.

Comparative study of the analysis of seized samples by GC-MS, 1H NMR and FT-IR spectroscopy within a Night-Time Economy (NTE) setting

Rapid analysis of surrendered or seized drug samples provides important intelligence for health (e.g. treatment or harm reduction), and custodial services. Herein, three in-situ techniques, GC-MS, 1H NMR and FT-IR spectroscopy, with searchable libraries, are used to analyse 318 samples qualitatively, using technique specific library-based searches, obtained over the period 24th – 29th August 2019. 259 samples were identified as consisting of a single component, of which cocaine was the most prevalent (n = 158). Median match scores for all three techniques were ≥ 0.84 and showed agreement except for metformin (n = 1), oxandrolone (identified as vitamin K by IR (n = 4)), diazepam (identified as zolpidem by FT-IR (n = 2)) and 2-Br-4,5-DMPEA (n = 1), a structural isomer of 2C-B identified as a polymer of cellulose (cardboard) by FT-IR. 51 samples were found to consist of two or more components, of which 49 were adulterated cocaine samples (45 binary and 4 tertiary samples). GC-MS identified all components present in the 49 adulterated cocaine samples, whereas IR identified only cocaine in 88 % of cases (adulterant only = 12 %). The breakdown for 1H NMR spectroscopy was all components identified (51 %), cocaine only (33 %), adulterant only (10 %), cocaine and one adulterant (tertiary mixtures only, 6 %).

Paraformaldehyde-coated electrochemical sensor for improved on-site detection of amphetamine in street samples

The increasing illicit production, distribution and abuse of amphetamine (AMP) poses a challenge for law enforcement worldwide. To effectively combat this issue, fast and portable tools for the on-site screening of suspicious samples are required. Electrochemical profile (EP)-based sensing of illicit drugs has proven to be a viable option for this purpose as it allows rapid voltammetric measurements via the use of disposable and low-cost graphite screen-printed electrodes (SPEs). In this work, a highly practical paraformaldehyde (PFA)-coated sensor, which unlocks the detectability of primary amines through derivatization, is developed for the on-site detection of AMP in seized drug samples. A potential interval was defined at the sole AMP peak (which is used for identification of the target analyte) to account for potential shifts due to fluctuations in concentration and temperature, which are relevant factors for on-site use. Importantly, it was found that AMP detection was not hindered by the presence of common diluents and adulterants such as caffeine, even when present in high amounts. When inter-drug differentiation is desired, a simultaneous second test with the same solution on an unmodified electrode is introduced to provide the required additional electrochemical information. Finally, the concept was validated by analyzing 30 seized AMP samples (reaching a sensitivity of 96.7 %) and comparing its performance to that of commercially available Raman and Fourier Transform Infrared (FTIR) devices.

Forensic Identification of Fentanyl and its Analogs by Electrochemical-Surface Enhanced Raman Spectroscopy (EC-SERS) for the Screening of Seized Drugs of Abuse

Reliable identification of fentanyl and fentanyl analogs present in seized drug samples is imperative to the safety of first responders and laboratory personnel and informs the future analysis process and handling procedures. The electrochemical-surface enhanced Raman spectroscopy (EC-SERS) method developed in this work allows the in-situ preparation of the SERS substrate providing a rapid, efficient, and accurate approach to detect fentanyl, even at low percent by weight concentrations common in seized drugs. Optimization of the electrochemical potentials suitable for the SERS substrate preparation and adsorption of the analyte was achieved using multi-pulse amperometric detection. This method demonstrated large enhancement of the SERS response. This method was applied to six fentanyl analogs with substitutions to the amide group, representing small changes in the fentanyl core structure. Identification of these analogs through differences in the EC-SERS spectra was evident. Interference studies incorporating analytes frequently encountered with fentanyl including heroin, cocaine, methamphetamine, naltrexone, and naloxone were assessed and found to offer limited to no interference. The limits of detection of the fentanyl compounds were in the low to mid nanograms per milliliter range, with the most sensitive compound detected at 10 ng/ml. Application of the method to simulated drug mixtures was performed to determine fit-for-purpose. In all mixtures with fentanyl as the minor contributor, fentanyl was correctly identified, including mixture samples comprised of 5 and 1% fentanyl. This approach represents the first in-situ EC-SERS analysis of fentanyl and its analogs and provides accurate and efficient screening for fentanyl in seized drug samples.

Rapid Determination of the 'Legal Highs' 4-MMC and 4-MEC by Spectroelectrochemistry: Simultaneous Cyclic Voltammetry and In Situ Surface-Enhanced Raman Spectroscopy.

The synthetic cathinones mephedrone (4-MMC) and 4-methylethcathinone (4-MEC) are two designer drugs that represent the rise and fall effect of this drug category within the stimulants market and are still available in several countries around the world. As a result, the qualitative and quantitative determination of ‘legal highs’, and their mixtures, are of great interest. This work explores for the first time the spectroelectrochemical response of these substances by coupling cyclic voltammetry (CV) with Raman spectroscopy in a portable instrument. It was found that the stimulants exhibit a voltammetric response on a gold screen-printed electrode while the surface is simultaneously electro-activated to achieve a periodic surface-enhanced Raman spectroscopy (SERS) substrate with high reproducibility. The proposed method enables a rapid and reliable determination in which both substances can be selectively analyzed through the oxidation waves of the molecules and the characteristic bands of the electrochemical SERS (EC-SERS) spectra. The feasibility and applicability of the method were assessed in simulated seized drug samples and spiked synthetic urine. This time-resolved spectroelectrochemical technique provides a cost-effective and user-friendly tool for onsite screening of synthetic stimulants in matrices with low concentration analytes for forensic applications.

Screening of seized drugs utilizing portable Raman spectroscopy and direct analysis in real time-mass spectrometry (DART-MS)

The continuous change of the drug landscape in the United States demands adaptation and incorporation of emerging analytical methods that preferably allow onsite screening but are also capable of supporting the analysis of seized drugs received at forensic laboratories across the country. Current methods employing color testing require the interaction of chemical reagents with powdered and liquid materials and can often yield inconclusive results, especially when the material is impure.Implementation of portable Raman spectroscopy can remove the need for direct interaction with solid and liquid specimens. In this study a portable, 785 nm, Raman spectroscopy system was employed for screening of seized drug samples, including mixtures. First, a panel of analytes comprised of 15 common drugs of abuse, 15 diluents, and 64 different mixtures comprising various ratios of analytes were used to measure bias, precision, and repeatability in accordance with United Nations Office on Drugs and Crime (UNODC) guidelines for handheld Raman field identification devices for seized material. Accuracy and precision through glass packaging was 91% and 90%; and through plastic was 89% and 88%, respectively for the diluents examined. A subset of the pure and mixture samples was then analyzed using direct analysis in real time mass spectrometry (DART-MS). Identification of analytes was performed manually by observing the [M+H]+ protonated molecule and conducting a library search of an in-house database. Using DART-MS, the drug analyte present in the sample was correctly identified 92% of the time using the library search feature. The presence of dimers and –OH losses were also observed for many of the analyte drugs of abuse. The combination of portable Raman spectroscopy and DART-MS data resulted in an overall accuracy of 96% for the detection of both drugs and diluents. The combined accuracy when analyzing authentic case samples was 83%, providing a rapid and accurate method for seized drug screening within drug chemistry laboratories.

Detection, discrimination and quantification of amphetamine, cathinone and nor-ephedrine regioisomers using benchtop 1 H and 19 F nuclear magnetic resonance spectroscopy.

Amphetamine and cathinone derivatives are abused recreationally due to the sense of euphoria they provide to the user. Methodologies for the rapid detection of the drug derivative present in a seized sample, or an indication of the drug class, are beneficial to law enforcement and healthcare providers. Identifying the drug class is prudent because derivatisation of these drugs, to produce regioisomers, for example, occurs frequently to circumvent global and local drug laws. Thus, newly encountered derivatives might not be present in a spectral library. Employment of benchtop nuclear magnetic resonance (NMR) could be used to provide rapid analysis of seized samples as well as identifying the class of drug present. Discrimination of individual amphetamine-, methcathinone-, N-ethylcathinone and nor-ephedrine-derived fluorinated and methylated regioisomers is achieved herein using qualitative automated 1 H NMR analysis and compared to gas chromatography-mass spectrometry (GC-MS) data. Two seized drug samples, SS1 and SS2, were identified to contain 4-fluoroamphetamine by 1 H NMR (match score median = 0.9933) and GC-MS (RRt = 5.42-5.43 min). The amount of 4-fluoroamphetamine present was 42.8%-43.4% w/w and 48.7%-49.2% w/w for SS1 and SS2, respectively, from quantitative 19 F NMR analysis, which is in agreement with the amount determined by GC-MS (39.9%-41.4% w/w and 49.0%-49.3% w/w). The total time for the qualitative 1 H NMR and quantitative 19 F NMR analysis is ~10 min. This contrasts to ~40 min for the GC-MS method. The NMR method also benefits from minimal sample preparation. Thus, benchtop NMR affords rapid, and discriminatory, analysis of the drug present in a seized sample.