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1.
Anal Chem ; 96(23): 9629-9635, 2024 Jun 11.
Article in English | MEDLINE | ID: mdl-38743697

ABSTRACT

Direct coupling of sample preparation with mass spectrometry (MS) can speed up analysis, enabling faster decision-making. In such combinations, where the analysis time is mainly defined by the extraction procedure, magnetic dispersive solid-phase extraction emerges as a relevant technique because of its rapid workflow. The dispersion and retrieval of the magnetic sorbent are typically uncoupled stages, thus reducing the potential simplicity. Stir bar sorptive dispersive microextraction (SBSDME) is a novel technique that integrates both stages into a single device. Its miniaturization (mSBSDME) makes it more portable and compatible with low-availability samples. This article reports the direct combination of mSBSDME and MS using a needle-based electrospray ionization (NESI) emitter as the interface. This combination is applied to determine tetrahydrocannabinol in saliva samples, a relevant societal problem if the global consumption rates of cannabis are considered. The coupling requires only the transference of the magnet (containing the sorbent and the isolated analyte) from the mSBSDME to the hub of a hypodermic needle, where the online elution occurs. The application of 5 kV on the needle forms an electrospray on its tip, transferring the ionized analyte to the MS inlet. The excellent performance of mSBSDME-NESI-MS/MS relies on the sensitivity (limits of detection as low as 2.25 ng mL-1), the precision (relative standard deviation lower than 15%), and the accuracy (relative recoveries ranged from 87 to 127%) obtained. According to the results, the mSBSDME-NESI-MS/MS technique promises faster and more efficient chemical analysis in MS-based applications.


Subject(s)
Dronabinol , Needles , Saliva , Spectrometry, Mass, Electrospray Ionization , Humans , Saliva/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Dronabinol/analysis , Solid Phase Microextraction/methods , Miniaturization , Limit of Detection
2.
Sensors (Basel) ; 24(10)2024 May 17.
Article in English | MEDLINE | ID: mdl-38794042

ABSTRACT

A rugged handheld sensor for rapid in-field classification of cannabis samples based on their THC content using ultra-compact near-infrared spectrometer technology is presented. The device is designed for use by the Austrian authorities to discriminate between legal and illegal cannabis samples directly at the place of intervention. Hence, the sensor allows direct measurement through commonly encountered transparent plastic packaging made from polypropylene or polyethylene without any sample preparation. The measurement time is below 20 s. Measured spectral data are evaluated using partial least squares discriminant analysis directly on the device's hardware, eliminating the need for internet connectivity for cloud computing. The classification result is visually indicated directly on the sensor via a colored LED. Validation of the sensor is performed on an independent data set acquired by non-expert users after a short introduction. Despite the challenging setting, the achieved classification accuracy is higher than 80%. Therefore, the handheld sensor has the potential to reduce the number of unnecessarily confiscated legal cannabis samples, which would lead to significant monetary savings for the authorities.


Subject(s)
Cannabis , Spectroscopy, Near-Infrared , Cannabis/chemistry , Cannabis/classification , Spectroscopy, Near-Infrared/methods , Discriminant Analysis , Least-Squares Analysis , Humans , Dronabinol/analysis
3.
J Chromatogr A ; 1727: 464994, 2024 Jul 19.
Article in English | MEDLINE | ID: mdl-38759461

ABSTRACT

This research aimed to support police forces in their battle against illicit drug trafficking by means of a multi-technique approach, based on gas chromatography. In detail, this study was focused on the profiling of volatile substances in narcotic Cannabis sativa L. flowering tops. For this purpose, the Scientific Investigation Department, RIS Carabinieri of Messina, provided 25 seized samples of Cannabis sativa L. The content of Δ9-tetrahydrocannabinol (THC), useful to classify cannabis plant as hemp (≤ 0.2 %) or as marijuana (> 0.2 %), was investigated. Essential oils of illicit drug samples were extracted using a microwave-assisted hydro-distillation (MAHD) system; GC-MS and GC-FID analytical techniques were used for the characterization of the terpenes and terpenoids fingerprint. Furthermore, the enantiomeric and carbon isotopic ratios of selected chiral compounds were investigated using a heart-cutting multidimensional GC (MDGC) approach. The latter exploited a combination of an apolar column in the first dimension, and a chiral cyclodextrin-based column in the second one, prior to parallel isotope-ratio mass spectrometry (C-IRMS) and MS detection. Finally, all the data were gathered into a statistical model, to demonstrate the existence of useful parameters to be used for the classification of seized samples.


Subject(s)
Cannabis , Distillation , Flowers , Gas Chromatography-Mass Spectrometry , Microwaves , Oils, Volatile , Cannabis/chemistry , Distillation/methods , Flowers/chemistry , Gas Chromatography-Mass Spectrometry/methods , Oils, Volatile/analysis , Oils, Volatile/chemistry , Terpenes/analysis , Dronabinol/analysis , Chromatography, Gas/methods
4.
Scand J Clin Lab Invest ; 84(2): 125-132, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38619215

ABSTRACT

This study investigated the effects of hexahydrocannabinol (HHC) and other unclassified cannabinoids, which were recently introduced to the recreational drug market, on cannabis drug testing in urine and oral fluid samples. After the appearance of HHC in Sweden in 2022, the number of posts about HHC on an online drug discussion forum increased significantly in the spring of 2023, indicating increased interest and use. In parallel, the frequency of false positive screening tests for tetrahydrocannabinol (THC) in oral fluid, and for its carboxy metabolite (THC-COOH) in urine, rose from <2% to >10%. This suggested that HHC cross-reacted with the antibodies in the immunoassay screening, which was confirmed in spiking experiments with HHC, HHC-COOH, HHC acetate (HHC-O), hexahydrocannabihexol (HHC-H), hexahydrocannabiphorol (HHC-P), and THC-P. When HHC and HHC-P were classified as narcotics in Sweden on 11 July 2023, they disappeared from the online and street shops market and were replaced by other unregulated variants (e.g. HHC-O and THC-P). In urine samples submitted for routine cannabis drug testing, HHC-COOH concentrations up to 205 (mean 60, median 27) µg/L were observed. To conclude, cannabis drug testing cannot rely on results from immunoassay screening, as it cannot distinguish between different tetra- and hexahydrocannabinols, some being classified but others unregulated. The current trend for increased use of unregulated cannabinols will likely increase the proportion of positive cannabis screening results that need to be confirmed with mass spectrometric methods. However, the observed cross-reactivity also means a way to pick up use of new cannabinoids that otherwise risk going undetected.


Subject(s)
Illicit Drugs , Substance Abuse Detection , Humans , Substance Abuse Detection/methods , Illicit Drugs/urine , Illicit Drugs/analysis , Sweden , Dronabinol/urine , Dronabinol/analysis , Dronabinol/analogs & derivatives , Cannabis/chemistry , Saliva/chemistry , Cannabinoids/urine , Cannabinoids/analysis , Cannabinol/analysis , Cannabinol/urine , Cross Reactions , Immunoassay/methods
5.
Chem Commun (Camb) ; 60(39): 5205-5208, 2024 May 09.
Article in English | MEDLINE | ID: mdl-38652014

ABSTRACT

This work developed an aptamer-dye complex as a label-free ratiometric fluorescence sensor for rapid analysis of THC and its metabolite in sewage samples. Integrated with a portable fluorescence capture device, this sensor exhibited excellent sensitivity with visualization of as low as 0.6 µM THC via naked-eye observation, and THC analysis can be accomplished within 4 min, which would be a complementary tool for quantifying THC in sewage samples to estimate cannabis consumption.


Subject(s)
Aptamers, Nucleotide , Dronabinol , Fluorescent Dyes , Sewage , Aptamers, Nucleotide/chemistry , Dronabinol/analysis , Dronabinol/chemistry , Fluorescent Dyes/chemistry , Sewage/analysis , Sewage/chemistry , Spectrometry, Fluorescence , Biosensing Techniques
6.
Psychopharmacology (Berl) ; 241(6): 1237-1244, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38407636

ABSTRACT

RATIONALE: As cannabis potency and cannabis use are increasing in newly legalized markets, it is increasingly important to measure and examine the effects of cannabinoid exposure. OBJECTIVES: The current study aims to assess how hair-derived cannabinoid concentrations - offering insight into three-month cumulative exposure - are associated with common self-report measures of cannabis use and cannabis use-related problems. METHODS: 74 near-daily dependent cannabis users self-reported their quantity of cannabis use, cannabis use-related problems, and estimated cannabis potency. Hair samples were provided to quantify Δ9-THC, CBD, and CBN using LC-MS/MS and THC-consumption was verified by analyzing THC-COOH in hair using GC-MS/MS. RESULTS: Cannabinoids were detectable in 95.95% of the hair samples from individuals who tested positive on a urine screen for cannabis. Δ9-THC concentrations were positively associated with measures of self-reported potency (relative potency, potency category, and perceived 'high'), but Δ9-THC, CBD, CBN concentrations and THC/CBD ratio were not associated with self-reported quantity of use. Self-reported potency, but not hair-derived concentrations, were associated with withdrawal and craving. Self-reported quantity of cannabis use, but not cannabinoid concentrations, were associated with cannabis use-related problems. CONCLUSIONS: The use of hair-derived cannabinoid quantification is supported for detecting cannabis use in near-daily users, but the lack of associations between hair-derived cannabinoid concentrations and self-report measures of use does not support the use of hair analyses alone for quantification of cannabinoid exposure. Further research comparing hair-derived cannabinoid concentrations with other biological matrices (e.g. plasma) and self-report is necessary to further evaluate the validity of hair analyses for this purpose.


Subject(s)
Cannabinoids , Hair , Self Report , Humans , Hair/chemistry , Male , Female , Adult , Cannabinoids/analysis , Young Adult , Marijuana Abuse , Substance Abuse Detection/methods , Dronabinol/analysis , Tandem Mass Spectrometry/methods , Middle Aged , Chromatography, Liquid/methods
7.
J Anal Toxicol ; 48(5): 372-379, 2024 Jun 11.
Article in English | MEDLINE | ID: mdl-38407251

ABSTRACT

With recent evolution of cannabis legalization around the world, cannabis edibles are booming, and determining their concentration in Δ9-tetrahydrocannabinol (Δ9-THC), the regulated psychoactive substance, remains a challenge for toxicology laboratories, which must prove whether the product has legal status or not. Cannabinoids are a large family of structurally similar and lipophilic molecules, requiring dedicated pre-analytical methods, as well as efficient chromatographic separation to differentiate cannabinoid isomers which are distinguished by their psychoactive properties and their legal status. Here, we present two independent cases of cannabis edibles, for which we performed analysis of homemade cannabis chocolate cakes and of the resins and herbs used for cooking. Quantitation was carried out with a new developed standard addition method, to avoid matrix effects and matrix-dependent calibration. Extraction by QuEChERs method, followed by targeted and non-targeted analysis by ultra-high performance liquid chromatography hyphenated to high resolution mass spectrometry (UHPLC-HRMS) allowed the identification of several phytocannabinoids, mainly Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD) and their acid precursors Δ9-THC acid (THCA) and CBD acid (CBDA). Δ9-THC was identified in significant concentrations (mg/g) in both edibles, even though one was prepared with CBD herb. This work highlights the need to analyze cannabis edibles, as well as the resins and herbs used in their preparation if it is homemade, and it proposes a reliable analytical method for toxicology laboratories.


Subject(s)
Cannabis , Dronabinol , Chromatography, High Pressure Liquid , Cannabis/chemistry , Dronabinol/analysis , Cannabinoids/analysis , Cannabidiol/analysis , Mass Spectrometry , Humans
8.
J Anal Toxicol ; 48(3): 165-170, 2024 Mar 28.
Article in English | MEDLINE | ID: mdl-38343275

ABSTRACT

In 2018, Canada introduced roadside oral fluid (OF) screening devices, called Approved Drug Screening Equipment (ADSE), as an investigative tool in impaired driving investigations to detect tetrahydrocannabinol (THC), cocaine and/or methamphetamine in drivers. In this work, we compare the detection and concentration of THC in blood samples collected from suspected impaired drivers that tested positive at the roadside for THC on an ADSE. The two ADSEs that were utilized were the Dräger DrugTest® 5000 (DDT) and the Abbott SoToxa™ (SoToxa), both configured with a THC OF concentration cut-off concentration of 25 ng/mL. Blood samples were screened for cannabinoids using immunoassay and positive results were followed up by confirmation/quantitation of THC by ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS-MS). A total of 230 cases were available where a blood sample was collected from a suspected impaired driver subsequent to a positive THC screen result on an ADSE. The blood samples were taken an average of 1.4 hours (range = 9 minutes to 3.2 hours) after the ADSE test. THC was confirmed in 98% of blood samples with concentrations across all samples ranging from not detected (cut = off 0.5 ng/mL) to greater than 20 ng/mL. Further, 90% of the blood samples had a THC concentration of 2.0 ng/mL (the lower per se limit in Canada) or greater. A positive ADSE test of a suspected impaired driver may predict that the driver has a detectable level of THC in their blood, and there is a high likelihood that the THC blood concentration is 2.0 ng/mL or higher. Hence, ADSE may be a useful tool for law enforcement and aid in the development of grounds to believe that a driver is operating a conveyance with a THC concentration exceeding Canadian per se limits.


Subject(s)
Dronabinol , Tandem Mass Spectrometry , Dronabinol/analysis , Tandem Mass Spectrometry/methods , Chromatography, Liquid , Drug Evaluation, Preclinical , Saliva/chemistry , Canada , Substance Abuse Detection/methods
9.
J Appl Physiol (1985) ; 136(3): 583-591, 2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38299223

ABSTRACT

Herein, we examine the human exercise response following cannabis inhalation, taking into consideration varied cannabinoid concentrations and different inhalation methods. A semirandomized crossover study design was used, with measures of perceived exertion and physiological responses to submaximal and maximal exercise. Participants (n = 14, 9 males 5 females) completed exercise after 1) smoking Δ-9-tetrahydrocannabinol (THC)-predominant cannabis (S-THC), 2) inhaling aerosol (vaporizing) from THC-predominant cannabis (V-THC), 3) inhaling aerosol from cannabidiol (CBD)-predominant cannabis (V-CBD), or 4) under control conditions. All exercise was performed on a cycle ergometer, with submaximal testing performed at 100 W followed by an evaluation of maximal exercise performance using an all-out 20-min time trial. Metabolism was characterized via the analysis of expired gases while subjective ratings of perceived exertion (RPE) were reported. During submaximal cycling, heart rate was higher during S-THC and V-THC compared with both control and V-CBD (all P < 0.02). During maximal exercise, V̇e was lower in V-THC compared with control, S-THC, and V-CBD (all P < 0.03), as was S-THC compared with control (P < 0.05). Both V̇o2 and RPE were similar between conditions during maximal exercise (both P > 0.1). Mean power output during the 20-min time trial was significantly lower in the S-THC and V-THC conditions compared with both control and V-CBD (all P < 0.04). Cannabis containing THC alters the physiological response to maximal and submaximal exercise, largely independent of the inhalation method. THC-containing cannabis negatively impacts vigorous exercise performance during a sustained 20-min effort, likely due to physiological and psychotropic effects. Inhalation of cannabis devoid of THC and primarily containing CBD has little physiological effect on the exercise response or performance.NEW & NOTEWORTHY Inhalation of cannabis containing THC alters physiological responses to both submaximal and maximal exercise and reduces mean power output during a 20-min time trial, regardless of whether it is inhaled as smoke or aerosol. In contrast, cannabis devoid of THC and predominantly containing CBD has no effect on physiological responses to exercise or performance.


Subject(s)
Cannabis , Dronabinol , Female , Humans , Male , Aerosols , Cannabidiol , Cannabinoids , Cannabis/chemistry , Cross-Over Studies , Dronabinol/analysis , Bicycling
10.
Planta Med ; 90(4): 316-332, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38387478

ABSTRACT

Concerns about health hazards associated with the consumption of trans-delta-8-tetrahydrocannabinol products were highlighted in public health advisories from the U. S. Food and Drug Administration and U. S. Centers for Disease Control and Prevention. Simple and rapid quantitative methods to determine trans-delta-8-tetrahydrocannabinol impurities are vital to analyze such products. In this study, a gas chromatography-flame ionization detection method was developed and validated for the determination of delta-8-tetrahydrocannabinol and some of its impurities (recently published) found in synthesized trans-delta-8-tetrahydrocannabinol raw material and included olivetol, cannabicitran, Δ 8-cis-iso-tetrahydrocannabinol, Δ 4-iso-tetrahydrocannabinol, iso-tetrahydrocannabifuran, cannabidiol, Δ 4,8-iso-tetrahydrocannabinol, Δ 8-iso-tetrahydrocannabinol, 4,8-epoxy-iso-tetrahydrocannabinol, trans-Δ 9-tetrahydrocannabinol, 8-hydroxy-iso-THC, 9α-hydroxyhexahydrocannabinol, and 9ß-hydroxyhexahydrocannabinol. Validation of the method was assessed according to the International Council for Harmonization guidelines and confirmed linearity with R2 ≥ 0.99 for all the target analytes. The limit of detection and limit of quantitation were 1.5 and 5 µg/mL, respectively, except for olivetol, which had a limit of detection of 3 µg/mL and a limit of quantitation of 10 µg/mL. Method precision was calculated as % relative standard deviation and the values were less than 8.4 and 9.9% for the intraday precision and inter-day precision, respectively. The accuracy ranged from 85 to 118%. The method was then applied to the analysis of 21 commercially marketed vaping products claiming to contain delta-8-tetrahydrocannabinol. The products analyzed by this method have various levels of these impurities, with all products far exceeding the 0.3% of trans-Δ 9-tetrahydrocannabinol limit for hemp under the Agriculture Improvement Act of 2018. The developed gas chromatography-flame ionization detection method can be an important tool for monitoring delta-8-tetrahydrocannabinol impurities in commercial products.


Subject(s)
Dronabinol , Dronabinol/analogs & derivatives , Resorcinols , Vaping , Dronabinol/analysis , Gas Chromatography-Mass Spectrometry/methods , Chromatography, Gas
11.
J AOAC Int ; 107(3): 493-505, 2024 May 02.
Article in English | MEDLINE | ID: mdl-38410076

ABSTRACT

While current analytical methodologies can readily identify cannabis use, definitively establishing recent use within the impairment window has proven to be far more complex, requiring a new approach. Recent studies have shown no direct relationship between impairment and Δ9-tetra-hydrocannabinol (Δ9-THC) concentrations in blood or saliva, making legal "per se" Δ9-THC limits scientifically unjustified. Current methods that focus on Δ9-THC and/or metabolite concentrations in blood, saliva, urine, or exhaled breath can lead to false-positive results for recent use due to the persistence of Δ9-THC well outside of the typical 3-4 h window of potential impairment following cannabis inhalation. There is also the issue of impairment due to other intoxicating substances-just because a subject exhibits signs of impairment and cannabis use is detected does not rule out the involvement of other drugs. Compounding the matter is the increasing popularity of hemp-derived cannabidiol (CBD) products following passage of the 2018 Farm Bill, which legalized industrial hemp in the United States. Many of these products contain varying levels of Δ9-THC, which can lead to false-positive tests for cannabis use. Furthermore, hemp-derived CBD is used to synthesize Δ8-THC, which possesses psychoactive properties similar to Δ9-THC and is surrounded by legal controversy. For accuracy, analytical methods must be able to distinguish the various THC isomers, which have identical masses and exhibit immunological cross-reactivity. A new testing approach has been developed based on exhaled breath and blood sampling that incorporates kinetic changes and the presence of key cannabinoids to detect recent cannabis use within the impairment window without the false-positive results seen with other methods. The complexity of determining recent cannabis use that may lead to impairment demands such a comprehensive method so that irresponsible users can be accurately detected without falsely accusing responsible users who may unjustly suffer harsh, life-changing consequences.


Subject(s)
Cannabis , Dronabinol , Substance Abuse Detection , Humans , Dronabinol/analysis , Substance Abuse Detection/methods , Cannabis/chemistry , Saliva/chemistry , Cannabidiol/analysis , Marijuana Abuse , Breath Tests/methods , Marijuana Use
12.
Electrophoresis ; 45(9-10): 948-957, 2024 May.
Article in English | MEDLINE | ID: mdl-38326083

ABSTRACT

Hemp and marijuana, both derived from Cannabis sativa L. (C. sativa), are subject to divergent legal regulations due to their different Δ9-tetrahydrocannabinol (Δ9-THC) contents. Cannabinoid synthase genes are considered the key enzymes that determine the chemical composition or chemotype of a particular cultivar. However, existing methods for crop type differentiation based on previous synthase gene theories have limitations in terms of precision and specificity, and a wider range of cannabis varieties must be considered when examining cannabis-based genetic markers. A custom next-generation sequencing (NGS) panel was developed targeting all synthase genes, including Δ9-THC acid synthase, cannabidiolic acid synthase, and cannabichromenic acid synthase, as well as the pseudogenes across diverse C. sativa samples, spanning reference hemp and marijuana, commercial hemp derivatives, and seized marijuana extracts. Interpretation of NGS data revealed a relationship between genotypes and underlying chemotypes, with the principal component analysis indicating a clear distinction between hemp and marijuana clusters. This differentiation was attributed to variations in both synthase genes and pseudogene variants. Finally, this study proposes a genetic cannabis classification method using a differentiation flow chart with novel synthase markers. The flow chart successfully differentiated hemp from marijuana with a 1.3% error rate (n = 147).


Subject(s)
Cannabis , High-Throughput Nucleotide Sequencing , Cannabis/genetics , Cannabis/chemistry , Cannabis/enzymology , High-Throughput Nucleotide Sequencing/methods , Dronabinol/analysis , DNA, Plant/genetics , DNA, Plant/analysis , Cannabinoids/analysis , Cannabinoids/metabolism , Intramolecular Oxidoreductases
13.
Ultrason Sonochem ; 103: 106766, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38271781

ABSTRACT

In this study, ultrasonic-ethanol pretreatment combined with AEE was developed for oil extraction from hemp seeds. The oil yield reached a maximum of 23.32 % at 200 W ultrasonic power and 30 min ultrasonic time, at this point, the degradation rate of Δ9-THC was 83.11 %. By determining the composition of hemp seed before and after pretreatment, it was shown that ultrasonic-ethanol pretreatment reduced the protein content of the raw material. An enzyme mixture consisting of pectinase and hemicellulase (1/1/1, w/w/w) was experimentally determined to be used, and the AEE extraction conditions were optimized using the Plackett-Burman design and the Box-Behnken. The optimal conditions were determined to be pH 5, total enzyme activity of 37,800 U/g, liquid-solid ratio of 10.4 mL/g, enzyme digestion temperature of 32 °C, enzymatic time of 189 min, and oil recovery of 88.38 %. The results of confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) showed that the emulsion formed during ultrasonic ethanol pretreatment was not uniformly distributed, and the droplets appeared to be aggregated; and the irregular pores of hemp seed increased after pretreatment. The contents of Δ9-THC and CBN in the extracted oil samples were 9.58 mg/kg and 52.45 mg/kg, respectively. Compared with the oil extracted by Soxhlet extraction (SE), the oil extracted by this experimental method was of better quality and similar in fatty acid composition.


Subject(s)
Cannabis , Plant Extracts , Cannabis/chemistry , Ultrasonics , Dronabinol/analysis , Ethanol/analysis , Seeds/chemistry , Water/chemistry , Plant Oils/chemistry
14.
Forensic Toxicol ; 42(1): 102-109, 2024 01.
Article in English | MEDLINE | ID: mdl-37603166

ABSTRACT

PURPOSE: Cannabis is regulated in many countries, and cannabis products are diversifying, which can hinder identification. Here, we report the seizure of a powder sample with a cannabis-like odor in a spice bottle labeled "nutmeg" and identification of the sample by chemical testing and cannabis DNA testing. METHODS: The sample was observed under a microscope, extracted with methanol, and analyzed by gas chromatography-mass spectrometry (GC-MS). The chemical profile of the seized powder was compared with that of nutmeg samples. Gas chromatography-flame ionization detection was used to estimate the total Δ9-tetrahydrocannabinol (Δ9-THC) concentration in the sample. A commercially available cannabis DNA testing kit was used to confirm the presence of cannabis plant DNA in the seized sample. RESULTS: The characteristics of cannabis in the seized powder were difficult to determine through microscopic observation alone. GC-MS analysis identified ß-caryophyllene (an aromatic component of cannabis) and five cannabinoids unique to cannabis, including Δ9-THC. No common compounds were identified in the seized powder or nutmeg samples. The total Δ9-THC concentration in the sample was very high (approximately 47% by weight). Cannabis DNA testing confirmed that the seized powder contained cannabis. CONCLUSIONS: The seized powder was found to be a processed product made from a finely pulverized resin-like cannabis concentrate. Our results indicate that combined chemical and DNA analysis should help identify cannabis-related samples in various forms.


Subject(s)
Cannabis , Hallucinogens , Cannabis/chemistry , Dronabinol/analysis , Powders , Gas Chromatography-Mass Spectrometry , Hallucinogens/analysis , Cannabinoid Receptor Agonists/analysis , DNA, Plant
15.
Drug Test Anal ; 16(2): 210-220, 2024 Feb.
Article in English | MEDLINE | ID: mdl-37343943

ABSTRACT

The analysis of cannabinoids in whole blood is usually done by traditional mass spectrometry (MS) techniques, after offline cleanup or derivatization steps which can be lengthy, laborious, and expensive. We present a simple, fast, highly specific, and sensitive method for the determination of Δ9 -tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), 11-hydroxy-Δ9 -tetrahydrocannabinol (11-OH-THC), and 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH) in 50 µL whole blood samples. After the addition of deuterated internal standards (IS) and a simple protein precipitation step, an online extraction of sample supernatants using turbulent flow chromatography (TurboFlow-Thermo Scientific) was carried out. Analytes were separated on a C18 analytical column and detected by LC-HRAM-Orbitrap-MS using a Thermo Scientific Q Exactive Focus MS system. MS detection was performed in polarity switching and selected ion monitoring (SIM) modes using five specific acquisition windows, at a resolution of 70,000 (FWHM). Total run time was about 10 min including preanalytical steps. Method validation was carried out by determining limit of detection (LOD), lower limit of quantitation (LLOQ), linearity range, analytical accuracy, intra-assay and interassay precision, carry-over, matrix effect, extraction recovery, and selectivity, for all analytes. Measurement uncertainties were also evaluated, and a decision rule was set with confidence for forensic purposes. The method may become suitable for clinical and forensic toxicology applications, taking advantage of the small matrix volume required, the simple and cost-effective sample preparation procedure, and the fast analytical run time. Performances were monitored over a long-term period and tested on 7620 driving under the influence of drugs (DUID) samples, including 641 positive samples.


Subject(s)
Cannabinoids , Driving Under the Influence , Cannabinoids/metabolism , Dronabinol/analysis , Mass Spectrometry , Cannabinol/analysis , Chromatography, Liquid/methods
16.
Drug Test Anal ; 16(2): 127-161, 2024 Feb.
Article in English | MEDLINE | ID: mdl-37269160

ABSTRACT

Since the early 2000s, there has been a turmoil on the global illicit cannabinoid market. Parallel to legislative changes in some jurisdictions regarding herbal cannabis, unregulated and cheap synthetic cannabinoids with astonishing structural diversity have emerged. Recently, semi-synthetic cannabinoids manufactured from hemp extracts by simple chemical transformations have also appeared as recreational drugs. The burst of these semi-synthetic cannabinoids into the market was sparked by legislative changes in the United States, where cultivation of industrial hemp restarted. By now, hemp-derived cannabidiol (CBD), initially a blockbuster product on its own, became a "precursor" to semi-synthetic cannabinoids such as hexahydrocannabinol (HHC), which appeared on the drug market in 2021. The synthesis and cannabimimetic activity of HHC were first reported eight decades ago in quest for the psychoactive principles of marijuana and hashish. Current large-scale manufacture of HHC is based on hemp-derived CBD extract, which is converted first by cyclization into a Δ8 /Δ9 -THC mixture, followed by catalytic hydrogenation to afford a mixture of (9R)-HHC and (9S)-HHC epimers. Preclinical studies indicate that (9R)-HHC has THC-like pharmacological properties. The animal metabolism of HHC is partially clarified. The human pharmacology including metabolism of HHC is yet to be investigated, and (immuno)analytical methods for the rapid detection of HHC or its metabolites in urine are lacking. Herein, the legal background for the revitalization of hemp cultivation, and available information on the chemistry, analysis, and pharmacology of HHC and related analogs, including HHC acetate (HHC-O) is reviewed.


Subject(s)
Cannabidiol , Cannabinoids , Cannabis , Hallucinogens , Illicit Drugs , Animals , Humans , United States , Cannabinoids/analysis , Cannabis/chemistry , Cannabinoid Receptor Agonists , Dronabinol/analysis
17.
Drug Test Anal ; 16(2): 174-186, 2024 Feb.
Article in English | MEDLINE | ID: mdl-37309060

ABSTRACT

Reports suggest that cannabis potency has dramatically increased over the last decade in the USA and Europe. Cannabinoids are the terpeno-phenolic compounds found in the cannabis plant and are responsible for its pharmacological activity. The two most prominent cannabinoids are delta-9-tetrahydrocannabinol (Δ9 THC) and cannabidiol (CBD). Cannabis potency is measured not only by the Δ9 THC levels but also by the ratio of Δ9 THC to other non-psychoactive cannabinoids, namely, CBD. Cannabis use was decriminalized in Jamaica in 2015, which opened the gates for the creation of a regulated medical cannabis industry in the country. To date, there is no information available on the potency of cannabis in Jamaica. In this study, the cannabinoid content of Jamaican-grown cannabis was examined over the period 2014-2020. Two hundred ninety-nine herbal cannabis samples were received from 12 parishes across the island, and the levels of the major cannabinoids were determined using gas chromatography-mass spectrometry. There was a significant increase (p < 0.05) in the median total THC levels of cannabis samples tested between 2014 (1.1%) and 2020 (10.2%). The highest median THC was detected in the central parish of Manchester (21.1%). During the period, THC/CBD ratios increased from 2.1 (2014) to 194.1 (2020), and there was a corresponding increase in the percent freshness of samples (CBN/THC ratios <0.013). The data show that a significant increase in the potency of locally grown cannabis has occurred in Jamaica during the last decade.


Subject(s)
Cannabidiol , Cannabinoids , Cannabis , Hallucinogens , Cannabis/chemistry , Dronabinol/analysis , Jamaica , Cannabinoids/analysis , Cannabidiol/analysis , Cannabinoid Receptor Agonists
18.
Behav Sleep Med ; 22(2): 150-167, 2024 Mar 03.
Article in English | MEDLINE | ID: mdl-37255232

ABSTRACT

INTRODUCTION: Cannabis is increasingly used to self-treat anxiety and related sleep problems, without clear evidence of either supporting or refuting its anxiolytic or sleep aid effects. In addition, different forms of cannabis and primary cannabinoids ∆9-tetrahydrocannabinol (THC) and cannabidiol (CBD) have differing pharmacological effects. METHODS: Thirty days of daily data on sleep quality and cannabis use were collected in individuals who use cannabis for mild-to-moderate anxiety (n = 347; 36% male, 64% female; mean age = 33 years). Participants self-reported both the form (flower or edible) and the ratio of THC to CBD in the cannabis used during the observation period. RESULTS: Individuals who reported cannabis use on a particular day also reported better sleep quality the following night. Moderation analyses showed that better perceived sleep after cannabis use days was stronger for respondents with higher baseline affective symptoms. Further, respondents who used cannabis edibles with high CBD concentration reported the highest perceived quality of sleep. CONCLUSIONS: Among individuals with affective symptoms, naturalistic use of cannabis was associated with better sleep quality, particularly for those using edible and CBD dominant products.


Subject(s)
Cannabidiol , Cannabis , Marijuana Smoking , Male , Humans , Female , Adult , Sleep Quality , Dronabinol/analysis , Dronabinol/pharmacology , Marijuana Smoking/psychology , Cannabidiol/therapeutic use , Cannabidiol/analysis , Cannabidiol/pharmacology , Anxiety/complications
19.
Addict Behav ; 150: 107930, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38091780

ABSTRACT

OBJECTIVE: Cannabis is widely used, including in early adolescence, with prevalence rates varying by measurement method (e.g., toxicology vs. self-report). Critical neurocognitive development occurs throughout adolescence. Given conflicting prior brain-behavior results in cannabis research, improved measurement of cannabis use in younger adolescents is needed. METHODS: Data from the Adolescent Brain Cognitive Development (ABCD) Study Year 4 follow-up (participant age: 13-14 years-old) included hair samples assessed by LC-MS/MS and GC-MS/MS, quantifying THCCOOH (THC metabolite), THC, and cannabidiol concentrations, and the NIH Toolbox Cognitive Battery. Youth whose hair was positive for cannabinoids or reported past-year cannabis use were included in a Cannabis Use (CU) group (n = 123) and matched with non-using Controls on sociodemographics (n = 123). Standard and nested ANCOVAs assessed group status predicting cognitive performance, controlling for family relationships. Follow-up correlations assessed cannabinoid hair concentration, self-reported cannabis use, and neurocognition. RESULTS: CU scored lower on Picture Memory (p = .03) than Controls. Within the CU group, THCCOOH negatively correlated with Picture Vocabulary (r = -0.20, p = .03) and Flanker Inhibitory Control and Attention (r = -0.19, p = .04), and past-year cannabis use was negatively associated with List Sorting Working Memory (r = -0.33, p = .0002) and Picture Sequence Memory (r = -0.19, p = .04) performances. CONCLUSIONS: Youth who had used cannabis showed lower scores on an episodic memory task, and more cannabis use was linked to poorer performances on verbal, inhibitory, working memory, and episodic memory tasks. Combining hair toxicology with self-report revealed more brain-behavior relationships than self-report data alone. These youth will be followed to determine long-term substance use and neurocognition trajectories.


Subject(s)
Cannabinoids , Cannabis , Hallucinogens , Marijuana Abuse , Adolescent , Humans , Tandem Mass Spectrometry , Chromatography, Liquid , Marijuana Abuse/diagnosis , Memory, Short-Term , Hair/chemistry , Cognition , Brain , Dronabinol/analysis
20.
J Sep Sci ; 47(1): e2300630, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37904320

ABSTRACT

Hemp-based materials have gained interest as alternative feed ingredients for livestock. However, safety concerns arise regarding the transfer of cannabinoids from the plant to the animals. Addressing these concerns requires the use of methods capable of detecting and quantifying cannabinoids in livestock. In this study, a fast and sensitive method was developed for quantification of cannabinoids and cannabinoid metabolites in cattle plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The extraction of cannabinoids from the plasma matrix was achieved by combining the Captiva Enhanced Matrix Removal-Lipid clean-up and salting-out assisted liquid-liquid extraction procedure. The developed method underwent validation using various analytical parameters, and the results demonstrated good accuracy, precision, specificity, and high sensitivity. The method was applied to real plasma samples obtained from cattle fed hemp for 2 weeks, and successfully detected various cannabinoids, including delta-9-tetrahydrocannabinol. Furthermore, the study revealed that 7-carboxy cannabidiol, a metabolite of cannabidiol, was the predominant cannabinoid present in the cattle plasma throughout the feeding period, which could remain detectable for weeks after the hemp feeding had ended.


Subject(s)
Cannabidiol , Cannabinoids , Cannabis , Cattle , Animals , Cannabinoids/analysis , Cannabidiol/analysis , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Liquid Chromatography-Mass Spectrometry , Dronabinol/analysis
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