Validation of a Bioanalytical Method for the Determination of Synthetic and Natural Cannabinoids (New Psychoactive Substances) in Oral Fluid Samples by Means of HPLC-MS/MS (original) (raw)

Determination of synthetic and natural cannabinoids in oral fluid by solid-phase microextraction coupled to gas chromatography/mass spectrometry: A pilot study

Talanta, 2019

In 2008, several synthetic cannabinoids were detected in herbal smoking blends sold on websites and in the so-called "smart shops". These compounds, as well as new psychoactive substances, flooded the market of illicit drugs and are sold at street level. Development and validation of rapid analytical methods for the detection and quantification of synthetic cannabinoids in biological matrices are essential for the investigation of pharmacokinetic, pharmacodynamic and toxicological properties. In this pilot study, the potential of direct immersion solid-phase microextraction coupled to GC/MS for the determination of synthetic and natural cannabinoids in oral fluid was proved. Validation proved the suitability of the developed method for the determination of the analytes at trace levels in oral fluid: linearity was assessed in the LOQ-1000 ng/mL range, whereas a good precision was observed with CV below 20%. The method was then applied to more than 100 real oral fluid samples collected from a population of young students, showing a positivity rate of 3.8%.

Screening of 104 New Psychoactive Substances (NPS) and Other Drugs of Abuse in Oral Fluid by LC–MS-MS

Journal of Analytical Toxicology, 2020

New psychoactive substances (NPS) are a major public health problem, primarily due to the increased number of acute poisoning cases. Detection of these substances is a challenge. The aim of this research was to develop and validate a sensitive screening method for 104 drugs of abuse, including synthetic cannabinoids, synthetic cathinones, fentanyl analogues, phenethylamines and other abused psychoactive compounds (i.e., THC, MDMA, LSD and their metabolites) in oral fluid by liquid chromatography–tandem mass spectrometry (LC–MS-MS). The Quantisal™ oral fluid device was used to collect oral fluid samples. The oral fluid–elution buffer mixture (500-μL sample) was extracted with t-butyl methyl ether, and chromatographic separation was performed on a Raptor™ biphenyl column (100 × 2.1 mm ID, 2.7 μm), with a total run time of 13.5 min. Limits of detection were established at three concentrations (0.05, 0.1 or 1 ng/mL) for most analytes, except for acetyl norfentanyl and mescaline (5 ng/mL...

Rapid identification of synthetic cannabinoids in herbal samples via direct analysis in real time mass spectrometry

Rapid Communications in Mass Spectrometry, 2012

RATIONALE: Dozens of synthetic cannabinoid analogs purposefully meant to circumvent legal restrictions associated with controlled substances continue to be manufactured and promoted as producing 'legal highs'. These designer drugs are difficult to identify in conventional drug screens not only because routine protocols have not been developed for their detection, but also because their association with complex plant matrices during manufacture generally requires laborintensive extraction and sample preparation for analysis. To address this new and important challenge in forensic chemistry, Direct Analysis in Real Time Mass Spectrometry (DART-MS) is applied to the analysis of these designer drugs. METHODS: DART-MS was employed to sample synthetic cannabinoids directly on botanical matrices. The ambient ionization method associated with DART-MS permitted the analysis of solid herbal samples directly, without the need for extraction or sample preparation. The high mass resolution time-of-flight analyzer allowed identification of these substances despite their presence within a complex matrix and enabled differentiation of closely related analogs. RESULTS: DART-MS was performed to rapidly identify the synthetic cannabinoids AM-251 and JWH-015. For each cannabinoid, three hundred micrograms (300 mg) of material was easily detected within an excess of background matrix by mass. CONCLUSIONS: New variations of herbal blends containing a wide range of base components and laced with synthetic cannabinoids are being produced, making their presence difficult to track by conventional methods. DART-MS permits rapid identification of trace synthetic cannabinoids within complex biological matrices, with excellent sensitivity and specificity compared with standard methods.

Use of High-Resolution Accurate Mass Spectrometry to Detect Reported and Previously Unreported Cannabinomimetics in "Herbal High" Products

Journal of Analytical Toxicology, 2010

A range of "Herbal High" products were tested for synthetic cannabinoids (cannabinomimetics) to qualitatively determine and compare their individual and relative content. Liquid chromatography-high resolution accurate mass spectrometry was used to rapidly screen samples for a range of cannabinomimetics using mono-isotopic masses derived from the elemental composition of target analytes. A screening database of over 140 compounds was rapidly created. This approach, combined with further tandem mass spectrometric experiments, also facilitated the detection and identification of compounds for which reference materials were not available. Previously reported cannabinomimetics, including JWH-018 and CP47,497 and its homologues, were detected in varying relative proportions along with several tentatively identified unreported cannabinomimetics. In some countries, the decision has been made to include these substances within their drug control legislation, and other countries are considering similar action. The currently applied drug screening techniques are unlikely to be effective in providing scientific evidence to support their identification in seized products. The application of high-resolution accurate mass spectrometry offers a solution. In addition, the technology provides a relatively simple and quick method for screening products, building substance databases, and even identifying novel substances using a combination of accurate mass derived elemental composition and fragment ions combined with fragmentation prediction software.

Simultaneous quantification of Δ9-tetrahydrocannabinol, 11-nor-9-carboxy-tetrahydrocannabinol, cannabidiol and cannabinol in oral fluid by microflow-liquid chromatography–high resolution mass spectrometry

Journal of Chromatography A, 2013

a b s t r a c t 9 -Tetrahydrocannabinol (THC) is the primary target in oral fluid (OF) for detecting cannabis intake. However, additional biomarkers are needed to solve interpretation issues, such as the possibility of passive inhalation by identifying 11-nor-9-carboxy-THC (THCCOOH), and determining recent cannabis smoking by identifying cannabidiol (CBD) and/or cannabinol (CBN). We developed and comprehensively validated a microflow liquid chromatography (LC)-high resolution mass spectrometry method for simultaneous quantification of THC, THCCOOH, CBD and CBN in OF collected with the Oral-Eze ® and Quantisal TM devices. One milliliter OF-buffer solution (0.25 mL OF and 0.5 mL of Oral-Eze buffer, 1:3 dilution, or 0.75 mL Quantisal buffer, 1:4 dilution) had proteins precipitated, and the supernatant subjected to CEREX TM Polycrom TM THC solid-phase extraction (SPE). Microflow LC reverse-phase separation was achieved with a gradient mobile phase of 10 mM ammonium acetate pH 6 and acetonitrile over 10 min. We employed a Q Exactive high resolution mass spectrometer, with compounds identified and quantified by targeted-MSMS experiments. The assay was linear 0.5-50 ng/mL for THC, CBD and CBN, and 15-500 pg/mL for THCCOOH. Intra-and inter-day and total imprecision were <10.8%CV and bias 86.5-104.9%. Extraction efficiency was 52.4-109.2%, process efficiency 12.2-88.9% and matrix effect ranged from −86 to −6.9%. All analytes were stable for 24 h at 5 • C on the autosampler. The method was applied to authentic OF specimens collected with Quantisal and Oral-Eze devices. This method provides a rapid simultaneous quantification of THCCOOH and THC, CBD, CBN, with good selectivity and sensitivity, providing the opportunity to improve interpretation of cannabinoid OF results by eliminating the possibility of passive inhalation and providing markers of recent cannabis smoking.

Development and Validation of a Liquid Chromatography High-Resolution Mass Spectrometry (LC-HRMS) Bioanalytical Method for Quantifying Cannabinoids in Whole Blood: Application for Determining Recent Cannabis Use

Journal of AOAC INTERNATIONAL

Background Cannabis legalization is expanding rapidly throughout the United States, but there is no reliable means of establishing recent use. Objective To develop and validate a bioanalytical method for determination of Δ9-tetrahydrocannabinol (Δ9-THC), cannabinol, 11-hydroxy-Δ9-THC, 11-nor-9-carboxy-Δ9-THC, and 8β,11-dihydroxy-Δ9-THC in whole blood microsamples by liquid chromatography high-resolution mass spectrometry (LC-HRMS). Methods Cannabinoid extraction from whole blood was performed using a mixture of n-hexane/ethyl acetate (90:10, v/v). Chromatographic separation was performed with a C18 column using a binary mobile phase gradient of water and acetonitrile, each with 0.1% formic acid. Detection was performed by positive ion mode heated electrospray ionization with full scan MS on an Orbitrap mass spectrometer. A clinical study was performed in 30 subjects to identify recent cannabis use based on analysis of cannabinoids in blood samples up to 200 min post-smoking. Results...