Article

Detection of Delta9-tetrahydrocannabinolic acid A in human urine and blood serum by LC-MS/MS

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Abstract

Delta9-tetrahydrocannabinolic acid A (Delta9-THCA-A) is the precursor of Delta9-tetrahydrocannabinol (Delta9-THC) in hemp plants. During smoking, the non-psychoactive Delta9-THCA-A is converted to Delta9-THC, the main psychoactive component of marihuana and hashish. Although the decarboxylation of Delta9-THCA-A to Delta9-THC was assumed to be complete--which means that no Delta9-THCA-A should be detectable in urine and blood serum of cannabis consumers--we found Delta9-THCA-A in the urine and blood serum samples collected from police controls of drivers suspected for driving under the influence of drugs (DUID). For LC-MS/MS analysis, urine and blood serum samples were prepared by solid-phase extraction. Analysis was performed with a phenylhexyl column using gradient elution with acetonitrile. For detection of Delta9-THCA-A, the mass spectrometer (MS) (SCIEX API 365 triple-quadrupole MS with TurboIonSpray source) was operated in the multiple reaction monitoring (MRM) mode using the following transitions: m/z357 --> 313, m/z357 --> 245 and m/z357 --> 191. Delta9-THCA-A could be detected in the urine and blood serum samples of several cannabis consumers in concentrations of up to 10.8 ng/ml in urine and 14.8 ng/ml in serum. The concentration of Delta9-THCA-A was below the Delta9-THC concentration in most serum samples, resulting in molar ratios of Delta9-THCA-A/Delta9-THC of approximately 5.0-18.6%. Only in one case, where a short elapsed time between the last intake and blood sampling is assumed, the molar ratio was 18.6% in the serum. This indicates differences in elimination kinetics, which need to be investigated in detail.

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... 4 However, this decarboxylation is only partial and, therefore, THCA-A can be found, together with THC, in the oral fluid, serum, and urine of Cannabis consumers. [5][6][7] Since THCA-A does not seem to convert to THC in vivo and displays its own metabolic and elimination pathways, it was proposed as a marker capable of distinguishing between the use of Cannabis and prescription synthetic THC ( Marinol Ò ). 8,9 Contrary to THC, THCA-A does not elicit psychoactive effects in humans and, perhaps for this reason, its pharmacological value is often neglected. ...
... 7 Although its absorption from Cannabis smoke is expected to be minimal, THCA-A can be detected in serum, urine, and oral fluid of Cannabis consumers up to 8 h after smoking. 5,6 For this reason, THCA-A was also investigated as a potential biomarker of Cannabis use hoping that it could potentially allow for a more accurate estimation of the time of Cannabis consumption than THC-COOH or THC. 5 Also, detection of THCA-A could undoubtedly differentiate between the intake of Cannabis products and prescribed THC medication (i.e., Marinol), which contains only pure THC. However, THCA-A was found to have a partition coefficient similar to THC and THC-COOH, and its blood levels not to correlate with the degree of impairment stated in police and medical reports. ...
... 5,6 For this reason, THCA-A was also investigated as a potential biomarker of Cannabis use hoping that it could potentially allow for a more accurate estimation of the time of Cannabis consumption than THC-COOH or THC. 5 Also, detection of THCA-A could undoubtedly differentiate between the intake of Cannabis products and prescribed THC medication (i.e., Marinol), which contains only pure THC. However, THCA-A was found to have a partition coefficient similar to THC and THC-COOH, and its blood levels not to correlate with the degree of impairment stated in police and medical reports. ...
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Δ(9)-tetrahydrocannabinolic acid A (THCA-A) is the acidic precursor of Δ(9)-tetrahydrocannabinol (THC), the main psychoactive compound found in Cannabis sativa. THCA-A is biosynthesized and accumulated in glandular trichomes present on flowers and leaves, where it serves protective functions and can represent up to 90% of the total THC contained in the plant. THCA-A slowly decarboxylates to form THC during storage and fermentation and can further degrade to cannabinol. Decarboxylation also occurs rapidly during baking of edibles, smoking, or vaporizing, the most common ways in which the general population consumes Cannabis. Contrary to THC, THCA-A does not elicit psychoactive effects in humans and, perhaps for this reason, its pharmacological value is often neglected. In fact, many studies use the term "THCA" to refer indistinctly to several acid derivatives of THC. Despite this perception, many in vitro studies seem to indicate that THCA-A interacts with a number of molecular targets and displays a robust pharmacological profile that includes potential anti-inflammatory, immunomodulatory, neuroprotective, and antineoplastic properties. Moreover, the few in vivo studies performed with THCA-A indicate that this compound exerts pharmacological actions in rodents, likely by engaging type-1 cannabinoid (CB1) receptors. Although these findings may seem counterintuitive due to the lack of cannabinoid-related psychoactivity, a careful perusal of the available literature yields a plausible explanation to this conundrum and points toward novel therapeutic perspectives for raw, unheated Cannabis preparations in humans.
... Pour le THCCOOH et le 11-OH-THC, les échantillons montrent des concentrations variant de 3 à 100 ng/mL et de 0,3 à 10 ng/mL respectivement. En ce qui concerne les échantillons de sérum, la concentration en THC est généralement inférieure à 70 ng/mL [15,16]. La concentration en THCCOOH est habituellement comprise entre 5 et 300 ng/mL [15]. ...
... En ce qui concerne les échantillons de sérum, la concentration en THC est généralement inférieure à 70 ng/mL [15,16]. La concentration en THCCOOH est habituellement comprise entre 5 et 300 ng/mL [15]. ...
... D'après les valeurs trouvées dans la littérature pour des échantillons réels [15,24,25], les concentrations habituellement obtenues pour le THCCOOH total varient entre 1 et 250 ng/mL. Cependant, des échantillons peuvent atteindre une concentration en THCCOOH de plus de 2000 ng/mL [24]. ...
Article
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Depuis quelques annees, la spectrometrie de masse en tandem (MS/MS) ne cesse de gagner du terrain comme methode d'analyse en toxicologie forensique, notamment pour le dosage des cannabinoides. Couplee a la chromatographie liquide (LC) ou gazeuse (GC), elle permet l'identification fiable et le dosage rapide du THC, de son precurseur acide, et de ses principaux metabolites, y compris les glucuronides. Au cours de ces dix dernieres annees, un nombre significatif de publications sont parues sur ce sujet. L'objectif de cet article est de passer en revue les analyses par spectrometrie de masse en tandem des cannabinoides dans diverses matrices biologiques.
... Jung et al. concluded that ''the determination of D9-tetrahydrocannabinolic acid A (THCA-A) may prove interesting for estimation of the elapsed time between the last intake and blood sampling, if THCA-A shows a shorter elimination half-life time and a lower partition coefficient than D9-tetrahydrocannabinol (THC), and therefore might be detectable only very shortly after consumption of cannabis'' [5]. THCA-A is the biosynthetic precursor of THC in cannabis plants, without having psychotropic effects by itself [6]. ...
... THCA-A has attracted scientists' attention in recent years. THCA-A was detected in serum and urine of cannabis consumers [5]. Additionally, THCA-A was found in oral fluid up to 8 h after smoking marijuana [7]. ...
... HPLC-DAD, GC-MS and NMR were used for THCA-A analysis in cannabis plants [2]. THCA-A was determined in human urine and blood serum samples by LC-MS/MS [5,12]. LC-MS/MS and LC-TOF-MS analysis was used for the identification of THCA-A metabolites in rat's urine samples [8]. ...
... Recently, several liquid-chromatography / mass spectrometry (LC-MS) or liquidchromatography / tandem mass spectrometry (LC-MS/MS) methods have been reported for the analysis of THC and/or THC-COOH in serum, plasma or whole blood [5][6][7][8][9], oral fluid [10,11] or urine [3,7,8,[12][13]. An isolation step of analytes is often required prior to instrumental analysis, performed generally by liquid-liquid extraction (LLE) [5,10,13] or solid phase extraction (SPE) [3,4,6,8,9,11,12], but it is a time-consuming step that also increases the cost of the assay and can affect the recovery. ...
... Recently, several liquid-chromatography / mass spectrometry (LC-MS) or liquidchromatography / tandem mass spectrometry (LC-MS/MS) methods have been reported for the analysis of THC and/or THC-COOH in serum, plasma or whole blood [5][6][7][8][9], oral fluid [10,11] or urine [3,7,8,[12][13]. An isolation step of analytes is often required prior to instrumental analysis, performed generally by liquid-liquid extraction (LLE) [5,10,13] or solid phase extraction (SPE) [3,4,6,8,9,11,12], but it is a time-consuming step that also increases the cost of the assay and can affect the recovery. ...
... Recently, several liquid-chromatography / mass spectrometry (LC-MS) or liquidchromatography / tandem mass spectrometry (LC-MS/MS) methods have been reported for the analysis of THC and/or THC-COOH in serum, plasma or whole blood [5][6][7][8][9], oral fluid [10,11] or urine [3,7,8,[12][13]. An isolation step of analytes is often required prior to instrumental analysis, performed generally by liquid-liquid extraction (LLE) [5,10,13] or solid phase extraction (SPE) [3,4,6,8,9,11,12], but it is a time-consuming step that also increases the cost of the assay and can affect the recovery. ...
Article
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A new high-throughput liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the quantification of Delta(9)-tetrahydrocannabinol (THC)in suspect plant material and of its principal metabolite, 11-nor-9carboxy-Delta 9-tetrahydrocannabinol (THC-COOH), in human urine was developed and validated. The separation was performed on a Zorbax SB-C18 column under isocratic conditions using a mixture of 2 mM ammonium formate in water and methanol (17:83, v/v) for THC or acetonitrile (45:55, v/v) for THC-COOH at 40 degrees C with a flow rate of 1 mL/min. The detection was performed in MRM mode using an ion trap MS equipped with an APCI ion source using positive mode ionization for THC and negative mode ionization for THC-COOH. The plant samples (0.2 g) were extracted in methanol and diluted before the analysis. The urine samples were heated for 30 min at 80 degrees C with 85% phosphoric acid and centrifugated before the analysis. The method showed a good linearity (r > 0.994), precision and accuracy (< 8.5%) over the concentration ranges of 2-128 ng/mL for THC and 0.5-32 ng/mL for THC-COOH, respectively. The method is not expensive, requires a minimum time for sample preparation and has a run-time of 3.5 min for instrument analysis (retention times for THC and THC-COOH were 2.6 and 2.55 min, respectively). The developed and validated LC-MS/MS method is very simple and more rapid than other similar methods. It was successfully applied for the rapid quantification of THC in plant samples and of THC-COOH in human urine in forensic toxicology analyses.
... The Δ 9 -THC precursor tetrahydrocannabinolic acid A (THCA) was also considered to be probably useful for the estimation of the time of last cannabis consumption. 13 In the investigation presented, we analyzed a total of 355 plasma samples (previously tested positive for Δ 9 -THC and THC-COOH) to assess the detectability of 14 other cannabinoids (next to Δ 9 -THC and THC-COOH) which might be useful to evaluate recent cannabis exposure. ...
... In accordance with our findings, 7 of the 12 tested serum samples containing high concentrations of Δ 9 -THC were positive for THCA (LOD = 2.5 ng/mL). 13 A limitation of this study is that phase II metabolites such as Δ 9 -THC-glucuronide were not considered. As these metabolites might also be of value for the proof of recent cannabis exposure, their detection incidence should be investigated in future. ...
Article
To assess the informative value of determining (minor) cannabinoids in plasma of cannabis users, detection rates of 14 cannabinoids (next to Δ⁹ -THC and THC-COOH) were determined. 11-OH-THC, THCA, CBC, CBN, and CBD were the most frequent detectable cannabinoids. The dependency of cannabinoid detectability on the plasma Δ⁹ -THC concentration was examined. © 2019 John Wiley & Sons, Ltd.
... So far, related deuterated cannabinoids (e.g. THC-COOH-D 3 ) were used as an internal standard for quantification of THCA-A [5,6]. Due to structural differences between this labeled compound and the target cannabinoid acid, quantification of THCA-A was susceptible to matrix effects and therefore not robust [5]. ...
... THC-COOH-D 3 ) were used as an internal standard for quantification of THCA-A [5,6]. Due to structural differences between this labeled compound and the target cannabinoid acid, quantification of THCA-A was susceptible to matrix effects and therefore not robust [5]. Hence, a strategy for the synthesis of deuterated THCA-A was developed. ...
... During method development, we attempted to include 9 -tetrahydrocannabinolic acid A (THCAA), THC's biosynthetic precursor because it was previously detected in serum [27,28], plasma and whole blood [29] from cannabis users, and may serve as a marker of illicit or recent cannabis administration. During validation, however, intra-and inter-day precision were 13.6-28.6%, ...
... was observed. Deuterated THCAA is not commercially available; previous methods quantified THCAA utilizing mismatched deuterated internal standards [27,30], or a custom-synthesized THCAA-d 3 internal standard [28,29]. We attempted to utilize THCCOOH-d 9 as an internal standard because it eluted closest to THCAA in the chromatographic gradient. ...
Article
Full-text available
Identifying recent cannabis intake is confounded by prolonged cannabinoid excretion in chronic frequent cannabis users. We previously observed detection times ≤2.1 h for cannabidiol (CBD) and cannabinol (CBN) and THC-glucuronide in whole blood after smoking, suggesting their applicability for identifying recent intake. However, whole blood collection may not occur for up to 4 h during driving under the influence of drugs investigations, making a recent-use marker with a 6–8 h detection window helpful for improving whole blood cannabinoid interpretation. Other minor cannabinoids cannabigerol (CBG), Δ9-tetrahydrocannabivarin (THCV), and its metabolite 11-nor-9-carboxy-THCV (THCVCOOH) might also be useful. We developed and validated a sensitive and specific liquid chromatography-tandem mass spectrometry method for quantification of THC, its phase I and glucuronide
... In Rauchkondensaten wurde THCA-A zum ersten Mal von Dussy et al. (2006) nachgewiesen [11] . Mittels einer Rauchapparatur wurde gezeigt, dass beim Cannabiskonsumenten nachgewiesen [47] . Eine daran anknüpfende Metabolismus-Studie in Ratten zeigte, dass mit einer Verstoffwechselung in Analogie zum ...
... Aus früheren Arbeiten existierte bereits eine Probenaufarbeitungsmethode für die quantitative Analyse von THCA-A und die qualitative Analyse von THCA-A-Metaboliten in verschiedenen Matrices [47] . Dabei kam eine Festphasenextraktion über eine C18- Genauigkeit quantitativ auf THCA-A untersucht. ...
Thesis
Delta9-Tetrahydrocannabinolsäure A (THCA-A) ist die psychoaktiv nicht wirksame, biogenetische Vorläufersubstanz von delta9-Tetrahydrocannabinol (THC) und Hauptbestandteil der Cannabinoidfraktion in frischem Cannabis-Pflanzenmaterial. Seit ihrer ersten Identifizierung in Serum- und Urinproben (2006) steht THCA-A als mögliche Markersubstanz für einen erst kurz zurückliegenden Cannabiskonsum im Gespräch. Der erste Teil dieser Arbeit befasst sich mit der Synthese eines deuterierten Analogons von THCA-A zur Verwendung als interner Standard in der massenspektrometrischen Analytik. THCA-A-D3 wurde ausgehend von THC-D3 über eine Carboxylierungsreaktion mit Methyl-Magnesiumcarbonat (MMC) und anschließender Hydrolyse mit verdünnter Salzsäure dargestellt. Im zweiten Teil dieser Arbeit wird die Entwicklung und Validierung einer LC-MS/MS-Methode zur Quantifizierung von THCA-A in Serum beschrieben. Unter Verwendung der synthetisierten THCA-A-D3 als interner Standard und einer fraktionierten Proteinfällung wurden eine gute Reproduzierbarkeit und niedrige Nachweis- und Bestimmungsgrenzen erhalten. Die Validierung wurde nach den Richtlinien der Gesellschaft für Toxikologische und Forensische Chemie (GTFCh) durchgeführt. Die Ergebnisse einer Pilotstudie aus dem Jahre 2011 (intravenöse Applikation von 5 mg THCA-A) gaben Grund zu der Annahme, dass THCA-A (ähnlich wie THC) in einem tiefen Kompartiment im menschlichen Körper (Fettgewebe) akkumuliert. Zur Prüfung dieser Annahme wurde ein Selbstversuch mit einem Probanden durchgeführt. Nach oraler Gabe einer Einzeldosis und täglicher Mehrfachgabe einer Kapsel mit 50 mg THCA-A über einen Zeitraum von 30 Tagen wurden Konzentrations-Zeit-Profile für THCA-A in Serum erstellt und nach Durchführung computergestützter pharmakokinetischer Analysen (Kompartiment- und Nicht-Kompartimentanalyse) verschiedene pharmakokinetische Parameter bestimmt (unter anderem Eliminationshalbwertszeit, mittlere Verweilzeit und Akkumulationsrate). Diese ergaben keinen Hinweis auf eine ausgeprägte Akkumulation von THCA-A im menschlichen Körper. Nichtsdestotrotz scheidet THCA-A in Serum aufgrund des langen Nachweisfensters (~ 98 Stunden) als Markersubstanz für einen erst kurz zurückliegenden Cannabiskonsum aus. Der dritte Teil dieser Arbeit beschäftigt sich mit der Haaranalytik. Es wurde eine LC-MS/MS-Methode zur Quantifizierung von THCA-A, THC, Cannabinol (CBN) und Cannabidiol (CBD) in Haaren entwickelt und diese nach den Richtlinien der GTFCh validiert. Die analytische Methode wurde zur Klärung der Herkunft in der Praxis häufig auftretender hoher THCA-A-Konzentrationen (> 1000 pg/mg) in Haaren von Cannabis-Konsumenten herangezogen. Dazu wurden sowohl Haarproben aus der Studie zur Akkumulation von THCA-A nach Mehrfachgabe als auch Proben einer Studie zur externen Kontamination von Haaren durch den Seitenstromrauch eines Joints untersucht. Es zeigte sich, dass THCA-A nicht über den Blutkreislauf in das Haar eingelagert wird und auch die Kontamination durch Seitenstromrauch nur zu einem sehr geringen Teil zu den hohen THCA A-Konzentrationen in Konsumentenhaar beitragen kann. Deutlich wurde dies über den Vergleich der Verhältnisse von THCA-A zu THC in Studien- und Konsumentenhaar. Extraktionsversuche während der Validierung zeigten, dass die Berührung der Haare mit kontaminierten Fingern (zum Beispiel nach dem Rollen eines Joints) als Hauptquelle für hohe THCA A-Konzentrationen in Frage kommt. Des Weiteren war die Bestimmung des Kontaminationsgrades verschiedener Kopfbereiche von Interesse. Es zeigte sich, dass im Fall von THC der bevorzugte Ort der Probennahme für Haaruntersuchungen (Hinterhaupthöcker) mit am stärksten von einer Kontamination durch Seitenstromrauch betroffen sein kann. Für THCA-A konnten aufgrund der allgemein niedrigen Konzentrationen keine Aussagen zur Verteilung gemacht werden.
... Methods for the analysis of THC and THCA-A in cannabis plant material [1][2][3], vegetable oils [4] as well as biological matrices, such as urine [5], serum [5,6], and hair [7] and also together with CBD and CBN [8] have been reported. For forensic analysis the total THC content must be determined. ...
... Methods for the analysis of THC and THCA-A in cannabis plant material [1][2][3], vegetable oils [4] as well as biological matrices, such as urine [5], serum [5,6], and hair [7] and also together with CBD and CBN [8] have been reported. For forensic analysis the total THC content must be determined. ...
Article
An HPLC-DAD method for the quantitative analysis of Δ(9)-tetrahydrocannabinol (THC), Δ(9)-tetrahydrocannabinolic acid-A (THCA-A), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis products has been developed, fully validated and applied to analyse seized cannabis products. For determination of the THC content of plant material, this method combines quantitation of THCA-A, which is the inactive precursor of THC, and free THC. Plant material was dried, homogenized and extracted with methanol by ultrasonication. Chromatographic separation was achieved with a Waters Alliance 2695 HPLC equipped with a Merck LiChrospher 60 RP-Select B (5μm) precolumn and a Merck LiChroCart 125-4 LiChrospher 60 RP-Select B (5μm) analytical column. Analytes were detected and quantified using a Waters 2996 photo diode array detector. This method has been accepted by the public authorities of Switzerland (Bundesamt für Gesundheit, Federal Office of Public Health), and has been used to analyse 9092 samples since 2000. Since no thermal decarboxylation of THCA-A occurs, the method is highly reproducible for different cannabis materials. Two calibration ranges are used, a lower one for THC, CBN and CBD, and a higher one for THCA-A, due to its dominant presence in fresh plant material. As provider of the Swiss proficiency test, the robustness of this method has been tested over several years, and homogeneity tests even in the low calibration range (1%) show high precision (RSD≤4.3%, except CBD) and accuracy (bias≤4.1%, except CBN).
... wileyonlinelibrary.com/journal/dta by experienced professionals and generally an additive, such as fluoride or EDTA is added. Determination of phytocannabinoids is generally performed in whole blood (WB), [31] [32] [33] [34] [35] [36] [37] [38] [39] but also plasma (P) [12,40–46] and serum (S) [47] can be used to predict the time of the last THC exposure. [48] [49] Analyses are generally performed in 0.5–1 mL of sample, but some studies have been performed on micro-volumes, scaling down to 50 μL. ...
... Some authors have used LLE as an alternative tech- nique, [33] [35] [37] [44] or SPE without any previous pretreatment. [38] [40] [43] [47] In recreational users THC concentration in WB is generally lower than 50 ng mL -1 ; THC-COOH and THC-OH have been found in the ranges of 3–100 ng mL -1 and 0.3–10 ng mL -1 , respectively. In case of recent assumption, in P and S samples THC is generally more concentrated (up to 100 ng mL -1 ), and so are its metabolites compared to WB. [48] On this basis, predictive models based on WB/P ratio have been developed for the estimation of last cannabis exposure; to assess recent intake generally accepted WB/P ratio is 0.5. ...
Article
Over the last two decades, the role played by phytocannabinoids and endocannabinoids in medicine has gained increasing interest in the scientific community. Upon identification of the plant compound Δ(9) -tetrahydrocannabinol (THC) and of the endogenous substance anandamide (AEA), different methodological approaches and innovative techniques have been developed, in order to evaluate the content of these molecules in various human matrices. In this review, we discuss the analytical methods that are currently used for the identification of phytocannabinoids and endocannabinoids, and we summarize the benefits and limitations of these procedures. Moreover, we provide an overview of the main biological matrices that have been analyzed to date for qualitative detection and quantitative determination of these compounds. Copyright © 2013 John Wiley & Sons, Ltd.
... As discussed, Δ 9 -THCA and Δ 9 -THC readily oxidize into CBNA and CBN in the presence of oxygen and light during thermal decarboxylation or even just upon aging ( Fig. 6c) (Moreno-Sanz 2016;Pellati et al., 2018;Dussy et al. 2005) in the same way, during storage or during decarboxylation, Δ 9 -THC can also oxidize into an isomer known as Δ 8 -THC, which is an artifact of the aging process (Pellati et al., 2018). As decarboxylation is only partial, THCA can be found, together with Δ 9 -THC, in the oral fluid, serum, and urine of cannabis consumers (Dussy et al. 2005;Jung et al. 2007;Moore et al. 2007). This can be used forensically, as THCA does not convert to Δ 9 -THC in vivo, displaying its own metabolic and elimination pathways (Fig. 6c); consequently, the presence of THCA distinguishes between the use of plantbased cannabis and prescribed synthetic Δ 9 -THC, e.g., Marinol® (Jung et al. 2009;Raikos et al. 2014). ...
Article
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Cannabis has been integral to Eurasian civilization for millennia, but a century of prohibition has limited investigation. With spreading legalization, science is pivoting to study the pharmacopeia of the cannabinoids, and a thorough understanding of their biosynthesis is required to engineer strains with specific cannabinoid profiles. This review surveys the biosynthesis and biochemistry of cannabinoids. The pathways and the enzymes’ mechanisms of action are discussed as is the non-enzymatic decarboxylation of the cannabinoic acids. There are still many gaps in our knowledge about the biosynthesis of the cannabinoids, especially for the minor components, and this review highlights the tools and approaches that will be applied to generate an improved understanding and consequent access to these potentially biomedically-relevant materials. Graphical abstract
... An important consideration for the cannabinoid analysis is that cannabinoids from plants are effectively in a "prodrug" form, existing as cannabinolic acids that must be decarboxylated to their respective cannabinol form to have pharmacological effects. 2 This decarboxylation, for example, occurs while smoking; however, upon oral consumption, no CBDA or THCA present is converted to CBD or THC by enzymatic or other processes. 37,38 If the production and processing of CBD oils does not remove all THCA and CBDA, some THCA and CBDA might still be present in the final CBD oil products. To evaluate whether THCA and CBDA would decarboxylate to their respective cannabinol forms during the AgPS-MS analysis and would thus interfere with the quantification of the THC/ CBD ratio, standard solutions of THCA and CBDA were analyzed with AgPS-MS (Supporting Information, Figure S9), Analytical Chemistry pubs.acs.org/ac ...
Article
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The control over the amount of psychoactive THC (Δ-9-tetrahydrocannabinol) in commercial cannabidiol (CBD) products has to be strict. A fast and simple semiquantitative Ag(I)-impregnated paper spray mass spectrometric method for differentiating between THC and CBD, which show no difference in standard single-stage or tandem MS, was established. Because of a different binding affinity to Ag(I) ions, quasi-molecular Ag(I) adducts [THC + Ag]⁺ and [CBD + Ag]⁺ at m/z 421 and 423 give different fragmentation patterns. The product ions at m/z 313 for THC and m/z 353 and 355 for CBD can be used to distinguish THC and CBD and to determine their ratio. Quantification of THC/CBD ratios in commercial CBD oils was accomplished with a low matrix effect (−2.2 ± 0.4% for THC and −2.0 ± 0.3% for CBD). After simple methanol extraction (recovery of 87.3 ± 1.2% for THC and 92.3 ± 1.4% for CBD), Ag(I)-impregnated paper spray analysis was employed to determine this ratio. A single run can be completed in a few minutes. This method was benchmarked against the UHPLC-UV method. Ag(I)-impregnated paper spray MS had the same working range (THC/CBD = 0.001–1) as UHPLC-UV analysis (R² = 0.9896 and R² = 0.9998, respectively), as well as comparable accuracy (−2.7 to 14%) and precision (RSD 1.7–11%). The method was further validated by the analysis of 10 commercial oils by Ag(I)-impregnated paper spray MS and UHPLC-UV analysis. Based on the determined relative concentration ratios of THC/CBD and the declared CBD concentration, 6 out of 10 CBD oils appear to contain more THC than the Dutch legal limit of 0.05%.
... Cannabinoids are generally determined by chromatographic techniques in real samples such as hair [16][17][18][19], urine [20][21][22] and drugs [23], among others. Commercial hemp oils enriched by CBD were also analyzed by chromatographic techniques, although no specific analysis is required for these products [24]. ...
Article
In this work, we present the first results obtained for the electroanalytical determination of Cannabidiol (CBD), the major non-psychoactive cannabinoid of Cannabis sativa L. plant, and Cannabinol (CBN). To this purpose, Sonogel-Carbon-PEDOT material was employed due to the high conductivity provided by poly-(3,4-ethylenedioxythiophene) (PEDOT) combined with graphite powder. Electrochemical measurements were performed in different aqueous buffered solutions mixed with acetonitrile or ethanol to improve the solubility of the analyte. In particular, the ethanol/borate buffer solution 15:85 (v/v) provided a higher electrochemical response of CBD as well as a lower potential peak value. CBD was assayed in a concentration range from 1.59 to 19.1 μM, obtaining high sensitivity, 421 ± 26.1 μA/mM·cm² and low detection limit, 0.94 μM. CBN was also assayed, from 1.61 to 25.8 μM, obtaining 588 ± 13.6 μA/mM·cm² as sensitivity and 1.29 μM as limit of detection. Based on the experimental results, the electroanalytical method developed represents a tool for the rapid determination of these types of cannabinoids.
... Neither compound is psychoactive per se, but both of these carboxylated compounds can be transformed into THC and CBD, respectively, through a degradative process which occurs slowly upon storage, but rapidly upon heating (Grotenhermen, 2003;Moreno-Sanz, 2016). Under field conditions, and with mild processing and cold storage, the extent of this decarboxylation is only nominal (~2-5%), which means that more carboxylic acid than phenolic forms are to be found in the oral fluid, serum, and urine of raw cannabis medical consumers (Dussy, Hamberg, Luginbühl, Schwerzmann, & Briellmann, 2005;Jung, Kempf, Mahler, & Weinmann, 2007;Moore, Rana, & Coulter, 2007;Moreno-Sanz, 2016). However, substantial to complete in situ decarboxylation occurs under the conditions of smoking or in baked goods. ...
Article
Cannabis is a plant with a long history of human pharmacological use, both for recreational purposes and as a medicinal remedy. Many potential modern medical applications for cannabis have been proposed and are currently under investigation. However, its rich chemical content implies many possible physiological actions. As the use of medicinal cannabis has gained significant attention over the past few years, it is very important to understand phytocannabinoid dispositions within the human body, and especially their metabolic pathways. Even though the complex metabolism of phytocannabinoids poses many challenges, a more thorough understanding generates many opportunities, especially regarding possible drug-drug interactions (DDIs). Within this context, computer simulations are most commonly used for predicting substrates and inhibitors of metabolic enzymes. These predictions can assist to identify metabolic pathways by understanding individual CYP isoform specificities to a given molecule, which can help to predict potential enzyme inhibitions and DDIs. The reported in vivo Phase I and Phase II metabolisms of various phytocannabinoids is herein reviewed, accompanied by a parallel in silico analysis of their predicted metabolism, highlighting the clinical importance of such understanding in terms of DDIs and clinical outcomes.
... The more labile "A" form is the immediate precursor of any THC found in marijuana, which results from the decarboxylation of biogenetic THCA due to environmental effects, heat (i.e., drying, smoking, baking), long-term storage and alkaline/acidic conditions. This process is only variably complete, so when cannabis is smoked or heated, some THCA can be detected in the serum and urine of cannabis users [54]. It is worth noting that the less volatile inhaled THCA is mostly converted to its own metabolites rather than to THC in vivo [55]. ...
Article
Marijuana (i.e., cannabis) and its derivatives are considered the most commonly used of illicit drugs. Within the last two decades, phytocannabinoids and their synthetic analogues have emerged as potential medicines for the treatment of various disorders via targeting of the endocannabinoid system. Recently, some countries have legalized (medicinal/recreational) cannabis, which now allows for more research to be conducted. Accordingly, sensitive and specific analytical assays are required to identify and quantify these compounds in different human matrices. These analytical approaches were developed using mass spectrometric detection, where LC-MS/MS specifically has become the mainstay for the quantitative analysis of tetrahydrocannabinol and other cannabinoids. This is due to its superior selectivity and sensitivity, and ability to determine free and conjugate analytes within the same analysis. This tabular review of such methods is prefaced by a short overview of the major cannabinoids and some of their physiological actions.
... Nevertheless a few methods for quantification of THC and CBD in plasma can be found in the literature and the majority of them involves time-consuming preparative steps with solid-phase [20] or liquid-liquid [21] extractions or derivatization steps [22] or start from high plasma volumes [23]. Moreover, the majority of them is based on ESI, which is the most commonly used ionization technique for drug identification. ...
Article
Aim: Monitoring of blood levels of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) is necessary for optimization of administration of medical cannabis. We describe the validation of a ultra-HPLC-MS/MS method for quantifying THC and CBD from plasma and decoctions and its application for therapeutic drug monitoring.Materials & methods: Analyses were performed by using a TSQ Quantiva™ Triple Quadrupole coupled to a Ultimate 3000 UHPLC system with atmospheric pressure chemical ionization after sample preparation with a straightforward method with deuterated internal standards. Results: The method has been validated following EMA guidelines and is linear in plasma from 0.16 to 10 ng/ml for both THC and CBD and in decoctions from 4.7 to 600 ng/ml. Conclusion: Given the unpredictable pharmacokinetic behavior of THC and CBD in patients, monitoring of plasma concentrations is strongly recommended for patients under treatment with medical cannabis.
... The oral absorption of THCA-A and CBDA in humans has never been documented. When THCA-A is inhaled, it does not seem to convert to THC in vivo; it displays its own metabolic and elimination pathways [13] and can be found, together with THC, in the blood serum and urine of Cannabis consumers [14,15]. ...
Article
Full-text available
Purpose: The recent release of a medical cannabis strain has given a new impulse for the study of cannabis in Italy. The National Health Service advises to consume medical cannabis by vaporizing, in decoction or oil form. This is the first study that explores the pharmacokinetics and tolerability of a single oral dose of cannabis as decoction (200 ml) or in olive oil (1 ml), as a first step to improve the prescriptive recommendations. Methods: This is a single-center, open-label, two-period crossover study designed to assess the pharmacokinetics and tolerability of oral cannabis administered to 13 patients with medication overuse headache (MOH). A liquid chromatography tandem-mass spectrometry (LC-MS/MS) method was conducted for the quantification of THC, CBD, 11-OH-THC, THC-COOH, THC-COOH-glucuronide, THCA-A, and CBDA. Blood pressure, heart rate, and a short list of symptoms by numerical rating scale (NRS) were assessed. Results: Decoctions of cannabis showed high variability in cannabinoids content, compared to cannabis oil. For both preparations, THCA-A and CBDA were the most widely absorbed cannabinoids, while THC and CBD were less absorbed. The most important differences concern the bioavailability of THC, higher in oil (AUC0-24 7.44, 95% CI 5.19, 9.68) than in decoction (AUC0-24 3.34, 95% CI 2.07, 4.60), and the bioavailability of CBDA. No serious adverse events were reported. Conclusions: Cannabis decoction and cannabis oil showed different pharmacokinetic properties, as well as distinct consequences on patients. This study was performed in a limited number of patients; future studies should be performed to investigate the clinical efficacy in larger populations.
... Jung et al. evaluated the presence of THC-A and other THC metabolites in urine and serum of cannabis smokers suspected of driving under the influence [14]. Only two cases provided THC-A positive results in urine. ...
Article
Introduction: The acidic forms of cannabinoids, THC-A and CBD-A are naturally present in cannabis plants and preparations and are generally decarboxylated to the active compounds before the use (e.g. thermally decarboxylated through smoking). Hence, the identification of the acidic compounds in urine could be an evidence of cannabis ingestion rather than a passive exposure to smoke. This case report described a 15-month-old child that suffered an acute intoxication by accidental cannabis ingestion. It is important to assess the ingestion and to discriminate it from a passive exposure to better interpret the clinical findings and to establish the correct therapeutic procedure. Methods: Urine samples were simply diluted in deionized water and directly injected in the LC-MS/MS system. D3-THCCOOH was used as internal standard. Chromatographic separation of THCCOOH, THC-A and CBD-A was carried out in reversed phase on a c18 column. A triple quad in MRM negative mode was used to monitor the three analytes. Results and discussion: The developed LC-MS/MS method was simple and fast. A LOD of 3.0ng/mL and a LOQ of 10.0ng/mL were measured for the three compounds. The analytical procedure was validated accordingly to international guidelines. The two urine samples collected from the 15-month-old child at the hospitalization and after three days provided positive results for THCCOOH (130.0 and 10.0ng/mL respectively). THC-A was found only in the urine sample collected at the hospitalization (concentration: 70.0ng/mL). Conclusion: THC-A was detected and quantitated in a urine sample of a 15-month-old child.
... According to the Canadian Centre on Substance Abuse (CCSA), substance use played a primary role in approximately 34.2 % of fatal car crashes in 2010 2 . Due to an increase in such accidents, many methods have been developed to detect commonly abused prohibited substances in oral fluid [3][4][5][6][7][8] and urine 3,9,10 . These methods generally use a chromatographic step as a confirmatory test, which places a time restraint on the whole procedure. ...
Article
The analysis of oral fluid (OF) and urine samples to detect drug consumption has garnered considerable attention as alternative biomatrices. Efficient implementation of microextraction and ambient ionization technologies for rapid detection of target compounds in such biomatrices creates a need for biocompatible devices which can be implemented for in vivo sampling and easily interfaced with mass spectrometry (MS) analyzers. This study introduces a novel solid-phase microextraction-transmission mode (SPME-TM) device made of poly(etheretherketone) (PEEK) mesh that can rapidly detect prohibited substances in biofluids via direct analysis in real-time tandem MS (DART-MS/MS). PEEK mesh was selected due to its biocompatibility, excellent resistance to various organic solvents, and its ability to withstand relatively high temperatures (≤350 °C). The meshes were coated with hydrophilic-lipophilic-balance particle-poly(acrylonitrile) (HLB-PAN) slurry. The robustness of the coated meshes was tested by performing rapid vortex agitation (≥3200 rpm) in LC/MS-grade solvents and by exposing them to the DART source jet stream at typical operational temperatures (∼250-350 °C). PEEK SPME-TM devices proved to be robust and were therefore used to perform ex vivo analysis of drugs of abuse spiked in urine and OF samples. Excellent results were obtained for all analytes under study; furthermore, the tests yielded satisfactory limits of quantitation (median, ∼0.5 ng mL-1), linearity (≥0.99), and accuracy (80-120%) over the evaluated range (0.5-200 ng mL-1). This research highlights plastic SPME-TM's potential usefulness as a method for rapidly screening for prohibited substances in on-site/in vivo scenarios, such as roadside or workplace drug testing, antidoping controls, and pain management programs.
... THCA-A can represent up to 90% of total THC content in the plant, it has about 70% conversion rate into THC when smoked (Dussy, Hamberg, Luginbuhl, Schwerzmann, & Briellmann, 2005): decarboxylation of THCA to THC is incomplete even at high temperatures in gas chromatography. Additionally, THCA can be detected in serum, urine, and oral fluid of cannabis consumers up to 8 h after smoking (Jung, Kempf, Mahler, & Weinmann, 2007). The cannabinoid acids do not produce any significant or documented psychotropic effects. ...
Chapter
The golden age of cannabis pharmacology began in the 1960s as Raphael Mechoulam and his colleagues in Israel isolated and synthesized cannabidiol, tetrahydrocannabinol, and other phytocannabinoids. Initially, THC garnered most research interest with sporadic attention to cannabidiol, which has only rekindled in the last 15 years through a demonstration of its remarkably versatile pharmacology and synergy with THC. Gradually a cognizance of the potential of other phytocannabinoids has developed. Contemporaneous assessment of cannabis pharmacology must be even far more inclusive. Medical and recreational consumers alike have long believed in unique attributes of certain cannabis chemovars despite their similarity in cannabinoid profiles. This has focused additional research on the pharmacological contributions of mono- and sesquiterpenoids to the effects of cannabis flower preparations. Investigation reveals these aromatic compounds to contribute modulatory and therapeutic roles in the cannabis entourage far beyond expectations considering their modest concentrations in the plant. Synergistic relationships of the terpenoids to cannabinoids will be highlighted and include many complementary roles to boost therapeutic efficacy in treatment of pain, psychiatric disorders, cancer, and numerous other areas. Additional parts of the cannabis plant provide a wide and distinct variety of other compounds of pharmacological interest, including the triterpenoid friedelin from the roots, canniprene from the fan leaves, cannabisin from seed coats, and cannflavin A from seed sprouts. This chapter will explore the unique attributes of these agents and demonstrate how cannabis may yet fulfil its potential as Mechoulam's professed “pharmacological treasure trove.”
... Smoking converts approximately 70% of the precursor to active THC (Dussey et al., 2005), documented by detection of the precursor in biological specimens following active or passive cannabis exposure (Auwärter et al., 2010;Jung et al., 2007;Moore et al., 2007;Moosmann et al., 2014;Raikos et al., 2014). Given that baking occurs at a lower temperature than smoking, we increased the conversion of the precursor to THC by including an additional step that baked the cannabis for extra time (based on personal communication with Dr. Ryan Vandrey who also conducted an edible cannabis study). ...
Article
Full-text available
Background: Although smoking is the most common cannabis administration route, vaporization and consumption of cannabis edibles are common. Few studies directly compare cannabis' subjective and physiological effects following multiple administration routes. Methods: Subjective and physiological effects, and expired carbon monoxide (CO) were evaluated in frequent and occasional cannabis users following placebo (0.001% Δ(9)-tetrahydrocannabinol [THC]), smoked, vaporized, and oral cannabis (6.9% THC, ∼54mg). Results: Participants' subjective ratings were significantly elevated compared to placebo after smoking and vaporization, while only occasional smokers' ratings were significantly elevated compared to placebo after oral dosing. Frequent smokers' maximum ratings were significantly different between inhaled and oral routes, while no differences in occasional smokers' maximum ratings between active routes were observed. Additionally, heart rate increases above baseline 0.5h after smoking (mean 12.2bpm) and vaporization (10.7bpm), and at 1.5h (13.0bpm) and 3h (10.2bpm) after oral dosing were significantly greater than changes after placebo, with no differences between frequent and occasional smokers. Finally, smoking produced significantly increased expired CO concentrations 0.25-6h post-dose compared to vaporization. Conclusions: All participants had significant elevations in subjective effects after smoking and vaporization, but only occasional smokers after oral cannabis, indicating partial tolerance to subjective effects with frequent exposure. There were no differences in occasional smokers' maximum subjective ratings across the three active administration routes. Vaporized cannabis is an attractive alternative for medicinal administrations over smoking or oral routes; effects occur quickly and doses can be titrated with minimal CO exposure. These results have strong implications for safety and abuse liability assessments.
... THC and CBD) due to heat, auto-oxidation, and light. While most common extraction and delivery methods of cannabis employ heat sufficient to convert most cannabinoids into their neutral form [45], decarboxylation is often incomplete and trace amounts of acidic cannabinoids can be found in the bodily fluids of cannabis consumers [46]. Certain delivery methods that have a long history of therapeutic use, such as cannabis tea, maintain the predominantly acidic state of cannabinoids [47]. ...
... The THC precursor, THCAA, was evaluated as a marker for differentiating illicit cannabis intake from licit THC pharmacotherapy [23]. THCAA concentrations were monitored in plasma and whole blood [24], blood serum and urine [25], and oral fluid [26,27]. THCAA also was demonstrated in hair after cannabis consumption, passive exposure, and external contamination [28,29]. ...
Article
A comprehensive cannabinoid urine quantification method may improve clinical and forensic result interpretation and is necessary to support our clinical research. A liquid chromatography tandem mass spectrometry quantification method for ∆9-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THCCOOH), ∆9-tetrahydrocannabinolic acid (THCAA), cannabinol (CBN), cannabidiol (CBD), cannabigerol (CBG), ∆9-tetrahydrocannabivarin (THCV), 11-nor-9-carboxy-THCV (THCVCOOH), THC-glucuronide (THC-gluc), and THCCOOH-glucuronide (THCCOOH-gluc) in urine was developed and validated according to the Scientific Working Group on Toxicology guidelines. Sample preparation consisted of disposable pipette extraction (WAX-S) of 200 μL urine. Separation was achieved on a Kinetex C18 column using gradient elution with flow rate 0.5 mL/min, mobile phase A (10 mM ammonium acetate in water), and mobile phase B (15 % methanol in acetonitrile). Total run time was 14 min. Analytes were monitored in both positive and negative ionization modes by scheduled multiple reaction monitoring. Linear ranges were 0.5–100 μg/L for THC and THCCOOH; 0.5–50 μg/L for 11-OH-THC, CBD, CBN, THCAA, and THC-gluc; 1–100 μg/L for CBG, THCV, and THCVCOOH; and 5–500 μg/L for THCCOOH-gluc (R 2 > 0.99). Analytical biases were 88.3–113.7 %, imprecisions 3.3–14.3 %, extraction efficiencies 42.4–81.5 %, and matrix effect −10 to 32.5 %. We developed and validated a comprehensive, simple, and rapid LC-MS/MS cannabinoid urine method for quantification of 11 cannabinoids and metabolites. This method is being used in a controlled cannabis administration study, investigating urine cannabinoid markers documenting recent cannabis use, chronic frequent smoking, or route of drug administration and potentially improving urine cannabinoid result interpretation.
... Es ist zudem nicht zu erwarten, dass bei Cannabis in absehbarer Zeit ähnlich umfassende Forschungsergebnisse vorliegen werden, wie dieses für die Rauch­ inhaltsstoffe des Tabaks gilt. So wurde erst vor einem Jahrzehnt die Bedeutung der beim Rauchen unvollständigen thermischen Decarboxylierung des THC­Vorläufers, der THC­Säure A, erkannt, die dann auch im Körper der Konsumenten nachgewiesen werden konnte [65]. THC­Säure A ist eine nicht psychoaktive Vorstufe von THC. ...
Article
In the context of the ongoing debate about the legalization of cannabis this article is focused in its general part on some common characteristics of intoxication itself and in its special part on the risks and adverse effects of the non-medical use of cannabis and synthetic cannabinoids. It is summarized that i) cannabis use is particularly prevalent among young adults, ii) the regular use of cannabis is decreasing among German teenagers (2011: 0.8 %), iii) 0.5 % of German adults are currently dependent on cannabis (DSM-IV), iv) the greatest health risks and psychosocial disabilities are expected to occur in regularly consuming young people and adult cannabis-dependents, v) both, the rate of the general disability and the health risks related to the use of cannabis are considerably lower than those related to the use of alcohol, vi) most psychic and physical sequelae of regular cannabis use are reversible in adults in the course of their abstinence; if the regular use had started in the adolescence or later; an earlier regular use is associated with sustaining impairments of executive functioning in adulthood vii) the risk of schizophrenia in regular cannabis users is doubled at least, viii) in modern cannabis strains the content of THC as well as cannabidiol had markedly increased and decreased, respectively, ix) the health risk subsequent to the consumption of synthetic cannabinoids is much higher than the health risk subsequent to the use of cannabis and is increasingly unpredictable, x) the number of emergency department visits involving the use of cannabis and synthetic cannabinoids are increasing and xi) children and young people are to be protected to prevent that substance use impedes their brain maturation and socialization/individual development by continuous consumption. In US-states, where cannabis had been legalized there exists a trend to increasing cannabis-related fatal traffic accidents, cyclic hyperemesis, emergency department visits and requests to Poison and Drug Centres. On the other hand, a decrease of deaths by overdose with opioide painkillers was observed.
... D 9 -Tetrahydrocannabinolic acid-A (THCA-A) is a precursor to THC in the cannabis plant. Upon heating, much of the THCA-A is decarboxylated to form THC. Jung et al. [120] demonstrated that THCA-A could be found in human urine and serum. THCA-A has also been suggested as a better urinary marker of recent can- a Sixty cannabis smokers resided on a closed research unit under 24-h monitoring for up to 30 d. ...
Article
Since 2004, when the World Anti-Doping Agency assumed the responsibility for establishing and maintaining the list of prohibited substances and methods in sport (i.e. the Prohibited List), cannabinoids have been prohibited in all sports during competition. The basis for this prohibition can be found in the World Anti-Doping Code, which defines the three criteria used to consider banning a substance. In this context, we discuss the potential of cannabis to enhance sports performance, the risk it poses to the athlete's health and its violation of the spirit of sport. Although these compounds are prohibited in-competition only, we explain why the pharmacokinetics of their main psychoactive compound, Delta(9)-tetrahydrocannabinol, may complicate the results management of adverse analytical findings. Passive inhalation does not appear to be a plausible explanation for a positive test. Although the prohibition of cannabinoids in sports is one of the most controversial issues in anti-doping, in this review we stress the reasons behind this prohibition, with strong emphasis on the evolving knowledge of cannabinoid pharmacology.
... As detailed in Section 1.4 ( Figure 1) a non-psychoactive ∆ 9 -THC precursor (∆ 9 -THCA-A) occurs in hemp. ∆ 9 -THCA-A may be absorbed by different routes, including the oral one, and has been found at trace levels in blood and urine from cannabis smokers (Jung et al., 2007). ...
Technical Report
The European Food Safety Authority (EFSA) was asked to deliver a scientific opinion on the risks for human health related to the presence of tetrahydrocannabinol (THC) in milk and other food of animal origin. THC, more precisely delta-9-tetrahydrocannabinol (Δ⁹-THC) is derived from the hemp plant Cannabis sativa. In fresh plant material, up to 90 % of total Δ⁹-THC is present as the non-psychoactive precursor Δ⁹-THC acid. Since few data on Δ⁹-THC levels in foods of animal origin were available, the Panel on Contaminants in the Food Chain (CONTAM Panel) estimated acute human dietary exposure to Δ⁹-THC combining different scenarios for the presence of Δ⁹-THC in hemp seed-derived feed materials. Acute exposure to Δ⁹-THC from the consumption of milk and dairy products ranged between 0.001 and 0.03 µg/kg body weight (b.w.) per day in adults, and 0.006 and 0.13 µg/kg b.w. per day in toddlers. From human data, the CONTAM Panel concluded that 2.5 mg Δ⁹-THC/day, corresponding to 0.036 mg Δ⁹-THC/kg b.w. per day, represents the lowest observed adverse effect level. By applying an overall uncertainty factor of 30, an acute reference dose (ARfD) of 1 μg Δ⁹-THC/kg b.w. was derived. The exposure estimates are at most 3 % and 13 % the ARfD, in adults and toddlers, respectively. The CONTAM Panel concluded that exposure to Δ⁹-THC via consumption of milk and dairy products, resulting from the use of hemp seed-derived feed materials at the reported concentrations, is unlikely to pose a health concern. A risk assessment resulting from the use of whole hemp plant-derived feed materials is currently not feasible due to a lack of occurrence data. The CONTAM Panel could also not conclude on the possible risks to public health from exposure to Δ⁹-THC via consumption of animal tissues and eggs, due to a lack of data on the potential transfer and fate of Δ⁹-THC.
... New models were suggested to identify recent or new cannabis use in occasional [8] and chronic frequent cannabis smokers [9]. There also were proposals to monitor unconjugated THCCOOH [10][11], Δ 9 -tetrahydrocannabinolic acid A [12], and THC-glucuronide [13], although these still need further research and validation. However, understanding cannabinoid stability is required to adequately interpret drug concentrations and apply these models or markers of recent use, especially if repeat analysis is requested months after initial testing. ...
Article
Full-text available
Analyte stability is an important factor in urine test interpretation, yet cannabinoid stability data are limited. A comprehensive study of Δ 9-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THCCOOH), cannabidiol, cannabinol, THC-glucuronide, and THCCOOH-glucuronide stabilities in authentic urine was completed. Urine samples after ad libitum cannabis smoking were pooled to prepare low and high pools for each study participant; baseline concentrations were measured within 24h at room temperature (RT), 4°C and −20°C. Stability at RT, 4°C and −20°C was evaluated by Friedman tests for up to 1 year. THCCOOH, THC-glucuronide, and THCCOOH-glucuronide were quantified in baseline pools. RT THCCOOH baseline concentrations were significantly higher than −20°C, but not 4°C baseline concentrations. After 1 week at RT, THCCOOH increased, THCCOOH-glucuronide decreased, but THC-glucuronide was unchanged. In RT low pool, total THCCOOH (THCCOOH +THCCOOH-glucuronide) was significantly lower after 1 week. At 4°C, THCCOOH was stable 2 weeks, THCCOOH-glucuronide 1 month and THC-glucuronide for at least 6 months. THCCOOH was stable frozen for 1 year, but 6 months high pool results were significantly higher than baseline; THC-glucuronide and THCCOOH-glucuronide were stable for 6 months. Total THCCOOH was stable 6 months at 4°C, and frozen 6 months (low) and 1 year (high). THC, cannabidiol and cannabinol were never detected in urine; although not detected initially, 11-OH-THC was detected in 2 low and 3 high pools after one week at RT. Substantial THCCOOH-glucuronide deconjugation was observed at RT and 4°C. Analysis should be conducted within 3 months if non-hydrolyzed THCCOOH or THCCOOH-glucuronide quantification is required.
... [8,9] . THCA-A wird durch Erhitzen -z.B. beim Rauchen -nur unvollständig zu THC decarboxyliert [10] (Abbildung 3) und wurde erstmals 2007 in Serum-, Urin-und Speichelproben von Cannabiskonsumenten nachgewiesen [11,12] . ...
Thesis
Die vorliegende Arbeit kann in drei Themenbereiche unterteilt werden. Der erste Teil befasst sich mit der Haaranalytik auf pflanzliche Cannabinoide und hierbei insbesondere mit den Haupteinlagerungswegen von THC, THCA-A und THC-COOH in das menschliche Haar, wobei insbesondere die Problematik der externen Kontamination behandelt wird. Anhand der durchgeführten Studien konnte gezeigt werden, dass der Anteil an THC, der tatsächlich über den Blutkreislauf in das Haar eingelagert wird, bisher erheblich überschätzt wurde. THCA-A wird ebenso wie THC praktisch ausschließlich über externe Kontamination in das Haar eingelagert. Die Studien bezüglich THC-COOH machen deutlich, dass bei engem Körperkontakt auch eine Übertragung auf andere Personen möglich ist. Dies stellt den eindeutigen Nachweis eines Cannabiskonsums selbst im Fall der Detektion von THC-COOH im Haar in Frage und kann insbesondere bei der Interpretation von Analysenergebnissen in Kinderhaar (Sorgerechtsfragestellungen) von erheblicher Bedeutung sein. Der zweite Teil der Dissertation befasst sich mit synthetischen Cannabinoiden. Mittels der im Rahmen dieser Arbeit entwickelten Flash-Chromatographiemethode ist es möglich synthetische Cannabinoide beim erstmaligen Erscheinen auf dem „Legal-High“-Markt unverzüglich aus der entsprechenden Räuchermischung zu isolieren und als Referenzmaterial einzusetzen. Des Weiteren konnte basierend auf der Untersuchung des Metabolismus von AM-2201 erstmalig anhand von Humanproben aufzeigt werden, dass es im Rahmen der Verstoffwechselung von synthetischen Cannabinoiden mit terminal fluorierter Pentylseitenkette zu einem metabolischen Austausch des Fluoratoms gegen eine Hydroxylgruppe kommt. Die Untersuchung von über 600 Räuchermischungsproben zeigte zum Teil erhebliche Schwankungen im Wirkstoffgehalt und beweist, dass es für die Konsumenten solcher Produkte nicht möglich ist, die Droge exakt und reproduzierbar zu dosieren. In der Haaranalytik auf synthetische Cannabinoide zeichnen sich ähnliche Probleme ab, wie sie im Rahmen dieser Arbeit bereits für pflanzliche Cannabinoide aufgezeigt wurden. Der dritte Teil der Arbeit beschäftigt sich mit Designer Benzodiazepinen. Im Rahmen der vorliegenden Arbeit konnten die ersten sieben Vertreter dieser neuen Gruppe Neuer Psychoaktiver Substanzen (NPS) umfassend chemisch-analytisch charakterisiert werden. Im Falle von Pyrazolam, Flubromazepam und Diclazepam wurden darüber hinaus erste pharmakokinetischen Daten erhoben. Die identifizierten Hauptmetabolisierungsschritte entsprechen denen der bekannten Benzodiazepine und bestehen vorwiegend in Hydroxylierungsreaktionen. Die extrem hohe pharmakologische Potenz von Stoffen wie z.B. Flubromazolam birgt neben der Gefahr der missbräuchlichen Einnahme das Risiko der unbemerkten Beibringung als „KO-Mittel“.
... In literature, several articles dealing with the analyses and determination of cannabinoids in plant material [4][5][6], urine [7,8], blood [8,9], oral fluid [8,10,11] as well as hair [12] using various chromatographic methods are available [13]. ...
... This sample loss presents a hurdle in standard material preparations, such as calibrators and QCs, leading o inaccuracy and reproducibility issues. A simple and effective solution to this conundrum was to prepare THC-COOH and its glucuronide in methanol or methanol-based solution (8,21,23). In the current study, to avoid any other potential compound adsorption to container surfaces, both positive and negative standards were prepared in 50% methanol. ...
Article
A novel LC-MS-MS assay that simultaneously detects and quantitates 78 drugs and metabolites was developed and validated for chronic pain management. Urine specimen was diluted and mixed with internal standards (ISs) before injected into LC-MS-MS. Seventy-two analytes were detected with positive electrospray ionization mode and the remaining six analytes with negative mode. Two separate gradient elution chromatographic programs were established with the same mobile phases on the same bi-phenyl HPLC column. The assay was linear for all analytes with linear regression coefficient ranging 0.994-1.000. The intra-assay precision was between 1.7 and 8.8% and inter-assay precision between 1.9 and 12.2%, with bias <20% for all but six analytes. All analytes in urine specimens were stable for 7 days at 4°C, and no significant matrix effect or carryover was observed. A suboptimal recovery rate (60.0-156.8%) was observed for six analytes, potentially due to the lack of available deuterated ISs, requiring comparison to a chemically different IS. Method comparison using patient and proficiency testing samples demonstrated that this assay was sensitive and accurate. The assay improves on currently existing assays by including glucuronide conjugates, allowing direct detection of metabolites that might otherwise be missed by existing methods. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
... This is owing to the accumulation of THC in fatty tissue facilitated by its lipophilic properties (9). Jung et al. (10) showed the detection of 9-tetrahydrocannabinolic acid and THC in human blood serum and urine using liquid chromatography-tandem mass spectrometry (LC/MS/MS). THC can also be detected in fingernails (11) and hair (12). ...
Article
The continuing rise in home and vehicular arson cases involving the use of ignitable liquids continues to be an area of concern for criminal and civil investigators. In this study, the compound-specific δ(13)C values of various components of four flammable household chemicals were measured using a single quadrupole mass spectrometer and an isotope ratio mass spectrometer as simultaneous detectors for a gas chromatograph. Whereas compound-specific carbon isotope ratios were able to discriminate between different sources of neat (pre-combustion) ignitable liquids, analyses of the post-combustion residues were problematic. Weathering caused by combustion resulted in a significant increase in the (13)C content of specific peaks relative to the neat liquids (i.e. less negative delta values) such that the isotopic comparison of pre- and post-combustion residues resulted in fractionation ranging from 0 to +10‰. Because of the current lack of understanding of isotopic fractionation during combustion, and because of problems encountered with co-elution in the more complex samples, compound-specific IRMS does not appear to be suitable for fire debris analysis. The comparison of non-combusted or non-weathered ignitable liquids is much more reliable, especially for relatively simple mixtures, and is best suited for exclusionary purposes until such time as a comprehensive database of samples is developed. Without a measure of the population variance, one cannot presently predict the false positive identification rate for the comparison of two ignitable liquids; i.e. the probability that two random ignitable liquid samples have indistinguishable isotope ratios.
... [23] Therefore, THCA-A can be detected in blood and urine samples of cannabis smokers. [24] Additionally it has been shown that THCA-A is not incorporated significantly into the hair through the bloodstream after oral intake of THCA-A daily doses of 10 mg for 30 consecutive days [25] (applied limit of detection: 50 pg/mg). In many forensic hair samples, however, the concentrations of THCA-A are considerably higher than the concentrations of THC. ...
Article
Condensation of marijuana smoke on the hair surface can be a source of an external contamination in hair analysis and may have serious consequences for the person under investigation. Δ9-tetrahydrocannabinolic acid A (THCA-A) is found in marijuana smoke and in hair analysis, but is not incorporated into the hair through the bloodstream. Therefore it might be a promising marker for external contamination of hair and could facilitate a more accurate interpretation of analytical results. In this study, three participants were exposed to the smoke of one joint every weekday over three weeks. Inhalation was excluded by an alternative breathing source. Hair samples were obtained up to seven weeks after the last exposure and analyzed for THCA-A, Δ9-tetrahydrocannabinol (THC) and cannabinol (CBN) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Additionally 30 hair samples from various regions of the head were obtained seven weeks after the exposure from one participant. The obtained results show that the degree of contamination depends on the hair length, with longer hair resulting in higher THC and CBN concentrations (1300 pg/mg and 530 pg/mg at the end of the exposure period) similar to the ones typically found after daily cannabis consumption. THCA-A could be detected in relatively low concentrations. Analysis of the distribution of the contamination showed that the posterior vertex region was affected most. The relatively low THCA-A concentrations in the samples suggest that most of the THCA-A found in forensic hair samples is not caused by sidestream marijuana smoke, but by other sources. Copyright © 2013 John Wiley & Sons, Ltd.
... Therefore, a fully validated and more sensitive analytical method, applying a deuterated internal standard (IS) for each analyte was needed. In previous studies, THC-COOH-D 3 was used as an IS for THCA-A, [17,18] because deuterated THCA-A is not commercially available. However, this was not an ideal solution, particularly with regard to matrix effects. ...
Article
For analysis of hair samples derived from a pilot study ('in vivo' contamination of hair by sidestream marijuana smoke), an LC-MS/MS method was developed and validated for the simultaneous quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), Δ9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD). Hair samples were extracted in methanol for 4 h under occasional shaking at room temperature, after adding THC-D(3) , CBN-D(3) , CBD-D(3) and THCA-A-D(3) as an in-house synthesized internal standard. The analytes were separated by gradient elution on a Luna C18 column using 0.1% HCOOH and ACN + 0.1% HCOOH. Data acquisition was performed on a QTrap 4000 in electrospray ionization-multi reaction monitoring mode. Validation was carried out according to the guidelines of the German Society of Toxicological and Forensic Chemistry (GTFCh). Limit of detection and lower limit of quantification were 2.5 pg/mg for THCA-A and 20 pg/mg for THC, CBN and CBD. A linear calibration model was applicable for all analytes over a range of 2.5 pg/mg or 20 pg/mg to 1000 pg/mg, using a weighting factor 1/x. Selectivity was shown for 12 blank hair samples from different sources. Accuracy and precision data were within the required limits for all analytes (bias between -0.2% and 6.4%, RSD between 3.7% and 11.5%). The dried hair extracts were stable over a time period of one to five days in the dark at room temperature. Processed sample stability (maximum decrease of analyte peak area below 25%) was considerably enhanced by adding 0.25% lecithin (w/v) in ACN + 0.1% HCOOH for reconstitution. Extraction efficiency for CBD was generally very low using methanol extraction. Hence, for effective extraction of CBD alkaline hydrolysis is recommended. Copyright © 2013 John Wiley & Sons, Ltd.
... More than 80 metabolites of THC have been identified [6]. In 2007, Jung et al. [7], who studied the metabolism of THC-A, reported 12 THC-A metabolites. We suggest that THC-gluc, THCCOOH-gluc, THC-A and other metabolites are also degraded, possibly into polymers, oxidized compounds and other cannabinoids that could not be detected with our analytical method. ...
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Background and purpose: T-type Ca channels (ICa ) regulate neuronal excitability and contribute to neurotransmitter release. The phytocannabinoids Δ9 -tetrahydrocannabinol and cannabidiol effectively modulate T-type ICa , but effects of other biologically active phytocannabinoids on these channels are unknown. We thus investigated the modulation of T-type ICa by low abundance phytocannabinoids. Experimental approach: A fluorometric (FLIPR) assay was used to investigate modulation of human T-type ICa (CaV 3.1, 3.2 and 3.3) stably expressed in FlpIn-TREx HEK293 cells. The biophysical effects of some compounds were examined using whole-cell patch clamp recordings from the same cells. Key results: In the FLIPR assay, all eleven phytocannabinoids tested modulated T-type ICa , with most inhibiting CaV 3.1 and CaV 3.2 more effectively than CaV 3.3. Cannabigerolic acid was the most potent inhibitor of CaV 3.1 (pIC50 6.1 ± 0.6) and CaV 3.2 (pIC50 6.4 ± 0.4); in all cases phytocannabinoid acids were more potent than their corresponding neutral forms. In patch clamp recordings, cannabigerolic acid inhibited CaV 3.1 and 3.2 with similar potency to the FLIPR assay, the inhibition was associated with significant hyperpolarizing shift in activation and steady state inactivation of these channels. In contrast, cannabidiol, cannabidivarin and cannabigerol only affected channel inactivation. Conclusion and implications: Modulation of T-type calcium channels is a common property of phytocannabinoids, which all increase steady state inactivation at physiological membrane potentials, with some also affecting channel activation. Thus, T-type ICa may be a common site of action for phytocannabinoids, and the diverse actions of phytocannabinoids on channel gating may provide insight into structural requirement for selective T-type ICa modulators.
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In forensic toxicology, high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) is increasingly used for the fast and sensitive measurement of a wide range of drugs. For our routine casework, a liquid chromatography atmospheric pressure chemical ionization-tandem mass spectrometry (LC-APCI-MS/MS) method for the quantification of Δ9-tetrahydrocannabinol (Δ9-THC), cannabinol (CBN) and cannabidiol (CBD) in hair was established and fully validated. Separation was achieved using a Kinetex® C18 column (100 mm × 2.1 mm, 100 Å, 1.7 μm, Phenomenex) at a flow rate of 0.5 mL/min. Measurements were performed on a QTrap 5,500 mass spectrometer (Sciex, Darmstadt, Germany). Unexpected signals were observed in authentic THC-positive hair samples. First, a signal with a slightly shifted retention time of THC whose origin could be assigned to the isomer Δ8-THC. Second, additional peaks exhibiting the same fragments as CBN and Δ9-THC but eluting at different retention times were detected. Spiking experiments and enhanced product ion scans (EPI) pointed to the origin of these additional signals as result of in-source decarboxylation of Δ9-tetrahydrocannabinolic acid A (Δ9-THCA-A) into Δ9-THC and further partial oxidation of Δ9-THC into CBN, respectively. Positive findings of Δ9-THCA-A in hair have been shown to derive from external contamination, therefore, the herein described artefacts may be used as indirect markers for external contamination.
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Cannabis products have recently regained much attention due to the high pharmacological potential of their cannabinoid content. In this review, the most widely used sample preparation strategies for the extraction of cannabinoids are described for the specific application to either plant materials or biological matrices. Several analytical techniques are described pointing out their respective advantages and drawbacks. In particular, chromatographic methods, such as TLC, GC and HPLC, are discussed and compared in terms of selectivity and sensitivity. Various detection methods are also presented based on the specific aim of the cannabinoids analysis. Lastly, critical considerations are mentioned with the aim to deliver useful suggestions for the selection of the optimal and most suitable method of analysis of cannabinoids in either biomedical or cannabis derived samples.
Chapter
Diabetes mellitus is characterized by hyperglycemia. However, dyslipidemia, oxidative stress, and inflammation are major risks in the development of diabetes and its complications. Δ⁹-Tetrahydrocannabinol (Δ⁹-THC) is the main constituent of Cannabis sativa. Δ⁹-THC is known as a natural phytocannabinoid and agonist of cannabinoid receptors. The cannabinoid 1 and cannabinoid 2 receptors (CB1 and CB2) are included in the endocannabinoid system. Recent studies have provided evidence that expressions and functions of cannabinoid receptors exist in the islets of Langerhans. Furthermore, according to studies, both the agonist and antagonist of cannabinoid receptors influence the regulation of food intake and energy homeostasis. Δ⁹-THC, a cannabinoid receptor agonist, may play a role on the effects of reactive oxygen species production, inflammation, insulin secretion, and lipids regulation. This review focused on the role of Δ⁹-THC in diabetes.
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▼ Im allgemeinen Teil dieses Artikels werden einige Charakteristika des Rausches aufgegriffen. Im speziellen Teil befasst sich der Artikel auf der Grundlage der aktuellen Literatur mit den Risiken und Nebenwirkungen des Konsums von nicht-medizinischem Cannabis und synthetischen Cannabinoiden vor dem Hintergrund der laufenden Legalisierungsdebatte um Cannabis. Es wird zusammengefasst, dass i) der Cannabiskonsum besonders weit verbreitet bei jungen Erwachsenen ist, ii) der regelmäßige Cannabiskonsum bei deutschen Teenagern abnimmt (2011: 0.8 %), iii) in Deutschland etwa 0,5 % der Erwachsenen abhängig (DSM-IV) von Cannabis sind, iv) die größten gesundheitlichen und psychosozialen Behinderungen bei regelmäßig konsumierenden Jugendlichen und Cannabisabhängigen zu erwarten sind, v) die allgemeine Behinderung und das gesundheitliche Risiko durch den Konsum von Cannabis wesentlich geringer als die allgemeine Behinderung und das gesundheitliche Risiko durch den Konsum von Alkohol sind, vi) die meisten psychischen und körperlichen Schäden in Verbindung mit regelmäßigem Cannabiskonsum im Erwachsenenalter im Verlauf der Abstinenz reversibel sind, wenn der regelmäßige Konsum ab der späten Jugend begonnen hat; ein früherer Beginn des regelmäßigen Konsums ist mit einer hartnäckigen Störung von Exekutivfunktionen im Erwachsenalter assoziiert, vii) sich das Risiko für eine Schizophrenie bei regelmäßigen Cannabiskonsumenten mindestens verdoppelt, viii) der THC-Gehalt in modernen Cannabiszüchtungen deutlich gestiegen während der Gehalt von Cannabidiol gesunken ist, ix) das gesundheitliche Risiko durch den Konsum von synthetischen Cannabinoiden viel höher als das durch den Konsum von Cannabis ist und zunehmend unkalkulierbar wird, x) die medizinischen Notfälle in Verbindung mit dem Konsum von Cannabis und synthetischen Cannabinoiden steigen und xi) Kinder und Jugendliche unbedingt vor dem Substanzkonsum geschützt werden müssen, um deren Hirnreifung bzw. Sozialisierung/Individualentwicklung durch fortlaufenden Konsum nicht ungünstig zu beeinflussen. In Staaten der USA, in denen Cannabis legalisiert wurde, gibt es einen ersten Trend zur Zunahme von tödlichen Verkehrsunfällen in Verbindung mit Cannabis, ansteigenden Beobachtungen von zyklischer Hyperemesis, häufigeren medizinischen Notfällen und Anfragen an Giftnotrufzentralen wegen Cannabisintoxikationen – allerdings andererseits auch eine Abnahme von Todesfällen durch Überdosierung opiathaltiger Schmerzmittel.
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The non-medical (recreational) use of cannabis is common particularly among young adults. In light of the ongoing legalization debate the clinical impact of physical and psychosocial consequences of regular recreational cannabis consumption should be presented. Health consequences appear to be more pronounced the earlier the regular recreational cannabis use had been started in the individual's development. There is an increasing demand from recreational cannabis users for medical treatment of cannabis-related complaints including the cannabis withdrawal syndrome. Physical sequelae such as chronic bronchitis, cyclical hyperemesis and fertility problems are usually reversible along with abstinence. The often debilitating cannabis-related mental and cognitive complaints respond on a qualified inpatient detoxification treatment with high effect sizes (Cohen's d 0.7 -1.4). The severity of the cannabis addiction benefits sustainably from psychotherapeutic approaches and individual psychosocial counseling (Cohen's d 0,5-1,2). Currently, the actual health hazard of recreational cannabis use was evaluated by addiction experts to be significantly lower than that of tobacco or alcohol use. © Georg Thieme Verlag KG Stuttgart · New York.
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A chromatographic methodology for Δ9-THC analysis in cosmetics is described. The HPLC-UV-Vis technique was employed for quantification of Δ9-THC in the 1.0 to 100 ppm working range using an optimized mobile phase consisting of methanol/water (4:1). In these experimental conditions, Δ9-THC presented a retention time of 23.7 minutes at the optimized wave-length of 209 nm. The analytical curve presented a linear correlation coefficient of 3.136x10-6 and the limits of detection and quantification were 0.746 and 2.487 ppm, respectively. This methodology was employed to quantify Δ9-THC in hemp oil-based cosmetics and it was possible to detect this cannabinol in concentrations ranging from 0.728 to 2.672 ppm.
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Non-coloured phenolic compounds were determined by high performance liquid chromatography in samples of fortified wines. A solid-phase extraction procedure was optimized for pretreatment of the sample, whose optimization used surface response methodology. The variables evaluated were sample volume, elution solvent volume and flow rate. Application of the surface response methodology revealed that the conditions for extraction were 5 mL of sample, 20 mL of elution solvent and flow rate 0.06 mL s-1. Verification tests gave percent recoveries in the interval between 71.06 and 119.40%. The optimized method was applied to samples of white and red fortified wines with good results for determination of gallic acid, (+) catechin, caffeic acid, p-coumaric acid, ferulic acid and quercetin, indicating the suitability of the model employed and the applicability of surface response methodology in optimization of the extraction conditions.
Thesis
Ziel der vorliegenden Arbeit war es, die pharmakokinetischen Eigenschaften von delta9-Tetrahydrocannabinolsäure A (THCA) und ihren Metabolismus im Menschen aufzuklären. Ausgangspunkt war die These, dass THCA – die nicht psychoaktive, biogenetische Vorläufersubstanz von THC in der Cannabispflanze – ein Marker für kurz zurückliegenden Cannabiskonsum sein könnte. Ein solcher Marker wäre hilfreich, um einen akuten Cannabiseffekt anhand von Blut- oder Urinproben nachzuweisen, was bis heute ein Problem in der forensischen Praxis darstellt. Kernstück der Arbeit war eine Humanstudie mit 16 Probanden, die THCA oral und intravenös erhielten. Dafür waren im Vorfeld einige Vorarbeiten nötig: Um auf Enzymaktivitäten beruhende Unterschiede in der Pharmakokinetik zu erkennen, wurde als erstes eine Cocktail-Phänotypisierungsmethode für die fünf CYP-Isoenzyme 1A2, 2C9, 2C19, 2D6 und 3A4 entwickelt. Neben der Auswahl der Testsubstanzen, der Phänotypisierungsindices und eines Probennahmeschemas umfasste dieser Schritt auch die Entwicklung und Validierung einer effektiven Aufbereitungsmethode und einer empfindlichen Analysenmethode. Die für die Humanstudie benötigte THCA wurde mittels Flash-Chromatographie aus Cannabisrohmaterial isoliert. Mit zwei voneinander unabhängigen Systemen konnte schließlich hochreine THCA (Reinheit > 98,5%) gewonnen werden, die zur Herstellung der Prüfpräparate diente. Als Grundlage für die intravenöse Applikation der lipophilen und thermoinstabilen THCA erwies sich eine parenterale Fettemulsion als geeignet. Letzter Schritt war die Entwicklung einer ESI-LC-MS/MS-Methode zum empfindlichen Nachweis von THCA und ihren Metaboliten sowie die Validierung der Quantifizierungsmethode für THCA. Im Fokus standen die Optimierung der Probenaufbereitung mittels Proteinfällung und eine kurze Analysendauer. Die Humanstudie selbst war in zwei Teile unterteilt: Im ersten Abschnitt erhielten die Probanden 10 mg THCA oral, im zweiten 5 mg intravenös und gaben jeweils über 96 h Blut- und Urinproben ab. Die Phänotypisierung erfolgte zwischen beiden Abschnitten. Mit einer Nachweisgrenze von 0,1 ng/ml stellte sich bei der Messung der Proben schließlich heraus, dass THCA bei den meisten Probanden bis zur letzten Serumprobe nachweisbar ist. Im Urin fanden sich nur minimale THCA-Mengen, eine Umwandlung in vivo von THCA zu THC im Körper fand nicht statt. Es erfolgte eine pharmakokinetische Analyse gemäß den Prinzipien der „compartmental“ und der „non-compartmental analysis“, mit der erstmals grundlegende pharmakokinetische Parameter der THCA (AUC, Clearance, Verteilungsvolumina, Halbwertzeiten sowie Makro- und Mikrokonstanten des Kompartimentmodells) bestimmt wurden. Dabei zeigte sich, dass die Pharmakokinetik von THCA am besten durch ein Drei-Kompartimentmodell beschrieben werden kann, was auf die Existenz eines tiefen Kompartiments deutet. Die meisten Parameter ähnelten prinzipiell denen von THC. Der einzige fundamentale Unterschied zu THC war die Plasmaclearance, die für THCA zehnfach geringer ist als für THC. Die orale Bioverfügbarkeit lag bei ~ 40 %. Es bestätigte sich, dass der Metabolismus von THCA analog zu THC verläuft. Hauptmetabolite im Serum sind 11-OH-THCA, THCA-8-on und THCA-COOH-Glucuronid, THCA-COOH sowie 9,10 Bis-OH-Hexahydrocannabinolsäure A (HHCA). Wie zu erwarten war, sind die Hauptmetabolite im Urin vor allem Glucuronide, wobei THCA-COOH-Glucuronid, 11-OH-THCA-Glucuronid, 8- und 4’-OH-THCA-COOH-Glucuronid die intensivsten Signale lieferten. Oft fand sich bis zur letzten abgegebenen Urinprobe wie auch im Serum unglucuronidierte 9,10-Bis-OH-HHCA. Zusätzlich wurde ein in vitro Experiment mit isolierten CYP-Isoenzymen durchgeführt, in dem sich herausstellte, dass vor allem die drei CYP 450-Isoenzyme 2C9, 3A4 und 2C19 beim Abbau von THCA eine Rolle spielen. In einer statistischen Analyse wurden schließlich die aus der Phänotypisierungsstudie ermittelten Indices mit den THCA-Clearances korreliert und untersucht, ob sich weitere Hinweise auf die abbauenden Isoenzyme ergeben. Für CYP 3A4 ließ sich ein statistisch signifikanter Zusammenhang zwischen Enzymaktivität und Clearance bestätigen, für alle anderen Kombinationen war dies jedoch nicht möglich. Die Hypothese, THCA könnte als Marker für kurzzeitig zurückliegenden Cannabiskonsum dienen, bestätigte sich nicht. Dieser Anwendung steht die Umverteilung in ein tiefes Kompartiment entgegen, was zu Akkumulation und langer Nachweisbarkeit führt. Hilfreich für ein anderes Konzept könnten jedoch einige THCA-Metaboliten werden. Dieses Konzept verfolgt den Ansatz, verschiedene Cannabisinhaltsstoffe und nachrangig gebildete Metaboliten zu screenen und daraus Schlüsse über Konsumzeitpunkt und letztendlich den Effekt zu ziehen. In dieses Substanzspektrum könnten Metaboliten wie THCA-Glucuronid, 8alpha- bzw. 8beta-OH-THCA, 8,11-Bis-OH-THCA sowie 8- bzw. 4’-OH-THCA-COOH-Glucuronid, die sich bei allen Probanden nur kurz nach der Applikation fanden, eingeschlossen werden.
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Recreational drugs (illicit drugs, human and veterinary medicines, legal highs, etc.) often contain lacing agents and adulterants which are not related to the main active ingredient. Serious side effects and even the death of the consumer have been related to the consumption of mixtures of psychoactive substances and/or adulterants, so it is important to know the actual composition of recreational drugs. In this work, a method based on flow injection analysis (FIA) coupled with high-resolution mass spectrometry (HRMS) is proposed for the fast identification of psychoactive substances in recreational drugs and legal highs. The FIA and HRMS working conditions were optimized in order to detect a wide range of psychoactive compounds. As most of the psychoactive substances are acid-base compounds, methanol-0.1 % aqueous formic acid (1:1 v/v) as a carrier solvent and electrospray in both positive ion mode and negative ion mode were used. Two data acquisition modes, full scan at high mass resolution (HRMS) and data-dependent tandem mass spectrometry (ddMS/HRMS) with a quadrupole-Orbitrap mass analyzer were used, resulting in sufficient selectivity for identification of the components of the samples. A custom-made database containing over 450 substances, including psychoactive compounds and common adulterants, was built to perform a high-throughput target and suspect screening. Moreover, online accurate mass databases and mass fragmenter software were used to identify unknowns. Some examples, selected among the analyzed samples of recreational drugs and legal highs using the FIA-HRMS(ddMS/HRMS) method developed, are discussed to illustrate the screening strategy used in this study. The results showed that many of the analyzed samples were adulterated, and in some cases the sample composition did not match that of the supposed marketed substance.
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A cross-over controlled administration study of smoked cannabis was carried out on occasional and heavy smokers. The participants smoked a joint (11 % Δ9-tetrahydrocannabinol (THC)) or a matching placebo on two different occasions. Whole blood (WB) and oral fluid (OF) samples were collected before and up to 3.5 h after smoking the joints. Pharmacokinetic analyses were obtained from these data. Questionnaires assessing the subjective effects were administered to the subjects during each session before and after the smoking time period. THC, 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THCCOOH) were analyzed in the blood by gas chromatography or liquid chromatography (LC)-tandem mass spectrometry (MS/MS). The determination of THC, THCCOOH, cannabinol (CBN), and Δ9-tetrahydrocannabinolic acid A (THC-A) was carried out on OF only using LC-MS/MS. In line with the widely accepted assumption that cannabis smoking results in a strong contamination of the oral cavity, we found that THC, and also THC-A, shows a sharp, high concentration peak just after smoking, with a rapid decrease in these levels within 3 h. No obvious differences were found between both groups concerning THC median maximum concentrations measured either in blood or in OF; these levels were equal to 1,338 and 1,041 μg/L in OF and to 82 and 94 μg/L in WB for occasional and heavy smokers, respectively. The initial WB THCCOOH concentration was much higher in regular smokers than in occasional users. Compared with the occasional smokers, the sensation of confusion felt by the regular smokers was much less while the feeling of intoxication remained almost unchanged. Figure Time profiles of THC, 11-OH-THC, and THCCOOH in whole blood for occasional (a) and heavy cannabis smokers (b)
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Cannabis is the most widely used illicit drug in the world. The pharmacological properties of Δ(9)-tetrahydrocannabinol also make it a promising molecule in the treatment of different pathologies. Understanding the PKs and PDs of this drug requires the determination of the concentration of Δ(9)-tetrahydrocannabinol and metabolites in biological matrices. For this purpose many analytical methodologies using mass spectrometric detection have been developed. In recent years, LC-MS/MS has become the gold standard in analysis of tetrahydrocannabinol and its metabolites due to the high selectivity and sensitivity, but above all, due to the ability to determine free and conjugate analytes in one run.
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Background: Blood and plasma cannabinoid stability is important for test interpretation and is best studied in authentic rather than fortified samples. Methods: Low and high blood and plasma pools were created for each of 10 participants after they smoked a cannabis cigarette. The stabilities of Δ(9)-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THCCOOH), cannabidiol (CBD), cannabinol (CBN), THC-glucuronide, and THCCOOH-glucuronide were determined after 1 week at room temperature; 1, 2, 4, 12, and 26 (±2) weeks at 4 °C; and 1, 2, 4, 12, 26 (±2), and 52 (±4) weeks at -20 °C. Stability was assessed by Friedman test. Results: Numbers of THC-glucuronide and CBD-positive blood samples were insufficient to assess stability. In blood, 11-OH-THC and CBN were stable for 1 week at room temperature, whereas THC and THCCOOH-glucuronide decreased and THCCOOH increased. In blood, THC, THCCOOH-glucuronide, THCCOOH, 11-OH-THC, and CBN were stable for 12, 4, 4, 12, and 26 weeks, respectively, at 4 °C and 12, 12, 26, 26, and 52 weeks at -20 °C. In plasma, THC-glucuronide, THC, CBN, and CBD were stable for 1 week at room temperature, whereas THCCOOH-glucuronide and 11-OH-THC decreased and THCCOOH increased. In plasma, THC-glucuronide, THC, THCCOOH-glucuronide, THCCOOH, 11-OH-THC, CBN, and CBD were stable for 26, 26, 2, 2, 26, 12, and 26 weeks, respectively, at 4 °C and 52, 52, 26, 26, 52, 52, and 52 weeks, respectively, at -20 °C. Conclusions: Blood and plasma samples should be stored at -20 °C for no more than 3 and 6 months, respectively, to assure accurate cannabinoid quantitative results.
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Two mathematical models are described for the prediction of time of marijuana use from the analysis of a single plasma sample for cannabinoids. The models were derived from cannabinoid data obtained from a controlled clinical study of acute marijuana smoking. Model I was based on plasma delta 9-tetrahydrocannabinol (THC) concentrations and Model II was based on the ratio of 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (THCCOOH) to THC in plasma. The two models were validated with cannabinoid data from nine published and unpublished clinical studies. The data included plasma samples obtained from infrequent and frequent marijuana smokers and after oral marijuana administration. Cannabinoid plasma concentrations had been determined by a variety of analytical methods. The accuracy of model prediction was evaluated by comparison of the predicted time of prior drug use to the actual time of exposure. Predictions of time of exposure were generally accurate but tended to overestimate time immediately after smoking and tended to underestimate later times. A second assessment of the validity of the models was made by determining if actual time of use was within the 95% confidence interval. Model I correctly predicted the time of exposure within the 95% confidence interval for 235 of 261 samples (90.0%), and Model II was correct in 232 of 260 samples (89.2%). These prediction models may be beneficial to forensic scientists in the interpretation of cannabinoid plasma levels.
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A fast method using automated solid-phase extraction (SPE) and short-column liquid-chromatography coupled to tandem mass-spectrometry (LC/MS/MS) with negative atmospheric-pressure chemical ionisation (APCI) has been developed for the confirmation of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) in urine samples. This highly specific method which combines chromatographic separation and MS/MS-analysis can be used for the confirmation of positive immunoassay results with a NIDA cut-off of 15ng/ml. The conjugates of THCCOOH were hydrolysed prior to SPE, and a standard SPE was performed using C18-SPE columns. No derivatisation of the extracts was needed as in GC/MS analysis, and the LC run-time was 6.5min by gradient elution with a retention time of 2.4min. Linearity of calibration was obtained in the range between 0 and 500ng/ml (correlation coefficient R2=0.998). Using linear regression (0–50ng/ml) the limit of detection (LOD) was 2.0ng/ml and the limit of quantitation (LOQ) was 5.1ng/ml; day-to-day reproducibility and precision were tested at 15 and 250ng/ml and were 13.4ng/ml±3.3% and 255.8ng/ml±4.5%, respectively.
Article
A comparative study was done in women and men of the effects of 9-tetrahydrocannabinol (9-THC), intravenously or orally, on dynamic activity, metabolism, excretion, and kinetics. In general no differences between the two sexes were observed. 9-THC is converted by microsomal hydroxylation to 11-hydroxy-9-THC (11-OH-9-THC), which is both a key intermediate for further metabolism to 11-nor-9-THC-9-carboxylic acid (11-nor-acid) by liver alcohol-dehydrogenase enzymes and a potent psychoactive metabolite. Major differences in the ratio of the concentration of 11-OH-9-THC to that of 9-THC in plasma were found after intravenous dosing (ratio 1:10 to 20) compared with oral administration (ratio 0.5 to 1:1). The final metabolic products are the 11-nor-acids and the related, more polar acids. Urinary excretion of 9-THC is restricted to acidic nonconjugated and conjugated metabolites. After 72 hr mean cumulative urinary excretion, noted for both routes and for both sexes, ranged from 13% to 17% of the total dose. After 72 hr the cumulative fecal excretion for both sexes after intravenous administration ranged from 25% to 30%; after oral administration the range was 48% to 53%. Metabolites were found in the feces in large concentration in the nonconjugated form; concentrations of 11-OH-9-THC were particularly noteworthy. Kinetics of 9-THC and metabolites were much the same for female and male subjects. For 9-THC, terminal-phase t½s for both sexes, irrespective of the route, ranged from 25 to 36 hr. A comparison of the results for AU C i dose (9-THC) after oral dosing with comparable data from intravenous administration indicated bioavailability of the order of 10% to 20% for both sexes. After intravenous 9-THC, large apparent volumes of distribution were noted (about 10 l/kg for both sexes).
Article
The limits of quantitation for cannabinoids in serum using automated solid-phase extraction (SPE) and gas chromatography-mass spectrometry (GC/MS) have been determined. Extraction was performed automatically using C18-cartridges, extracts were methylated and analysed with selected-ion monitoring (SIM). The limits of quantitation (LOQ) in serum were determined according to Euronorm EN 45001 and German Industrial Norm DIN 32645 and were 0.1 and 0.7 ng/ml for tetrahydrocannabinol, 0.5 and 0.7 ng/ml for 11-hydroxy-tetrahydrocannabinol and < 0.25 and 1.6 ng/ml for tetrahydrocannabinolcarboxylic acid, respectively. The use of these LOQ’s in forensic cases and the necessity of drug identification prior to quantitation are discussed in detail. Due to the newest amendment of the German traffic law (§24a “Straßenverkehrsgesetz”) the quest for nationwide standardisation of the forensic analysis of illicit drugs in serum or blood arises. By automation of several manual sample preparation steps we could get closer to this aim. Taking into account a margin of safety, a qualitative cut-off at 1 ng/ml THC and a limit of quantitation of 2 ng/ml THC in serum should be achievable in all forensic toxicology laboratories.
Article
Cannabinoid acids readily decarboxylate to the corresponding cannabinoid. Methods are available for the determination of delta 9-tetrahydrocannabinol (THC) and its acids (THCA) and published data on the levels of these compounds in cannabis are summarized. Using gas and liquid chromatography, fresh cannabis (64 samples) and cannabis resin (26 samples) from different countries were examined. Wide variations in the relative amounts of THCA and THC in cannabis were found. For cannabis resin, a wide range of values was also found (0.5: 1 to 6.1: 1), the lower values being in resins from the Indian sub-continent and the higher values in resins from the Mediterranean area. Total THC values were in the range 1.0 - 10.6% in cannabis and 6.0 - 12.5% in cannabis resin.
Article
The metabolism of delta 9-tetrahydrocannabinol (THC) and related cannabinoids in man has been studied in detail utilizing intravenous, oral, and smoking routes of administration. The general pattern of metabolism was the same in all studies involving THC and related cannabinoids. Microsomal hydroxylation allylic to the delta 9-THC double bond occurs, the major product resulting in formation of an 11-CH2OH moiety; minor hydroxylation occurs on the C-8 carbon. Nonmicrosomal oxidation of the resultant 11-OH-delta 9-THC to 11-nor-delta 9-THC-9-carboxylic acid and to other more polar acids generates the major terminal metabolic products. After oral administration, approximately equal quantities of THC and its highly active 11-hydroxymetabolite were formed, whereas the latter metabolite is a minor constituent after administration by intravenous or smoking routes. Initial pharmacokinetic analyses of the data show that the mean terminal-phase (beta-phase) plasma half-life after intravenous administration of THC was about 30 hours; after oral administration, it was 23 hours. No significant statistical difference was noted between men and women as to metabolic routes or plasma terminal-phase half-lives.
Article
A solvent programmed reversed-phase HPLC method with UV detection for the determination of delta9-tetrahydrocannabinol (THC) and delta9-tetrahydrocannabinolic acid A (THCA-A) in foods containing parts of hemp such as edible oil, herb-teas (infusion), herbal hemp or hempseed is presented. The THC peak is also detected by fluorescence. The detection limits with UV detection are 0.01 ng for THC and 0.05 ng for THCA-A and with fluorescence detection 0.1 ng for THC. The relative standard deviation under repeatability conditions of the chromatographic procedure is about 0.5% and that of the over-all analytical procedure for THC in vegetable oils 2% (concentration range of 10-100 mg/kg).
Article
A fast method using liquid-liquid extraction and HPLC/tandem-mass spectrometry (LC/MS/MS) was developed for the simultaneous detection of 11-Nor-Delta(9)-tetrahydrocannabinol-9-carboxylic acid beta-glucuronide (THC-COOH-glucuronide) and 11-Nor-Delta(9)-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in urine samples. This highly specific method, which combines chromatographic separation and MS/MS analysis, can be used for the confirmation of positive immunoassay results even without hydrolysis of the sample or derivatisation of extracts. Liquid-liquid extraction was optimised: with ethylacetate/diethylether (1:1, v/v) THC-COOH-glucuronide and THC-COOH could be extracted in one step. Molecular ions of the glucuronide (MH(+), m/z 521) and THC-COOH (MH(+), m/z 345) were generated using a PE/SCIEX turboionspray source in positive ionisation mode; specific fragmentation was performed in the collision cell of an API 365 triple-quadrupole mass spectrometer and yielded major fragments at m/z 345 (for THC-COOH-glucuronide) and m/z 327 as well as m/z 299 for both cannabinoids. Chromatographic separation was performed using a reversed-phase C8 column and gradient elution with 0.1% formic acid/1 mM ammonium formate and acetonitrile/0.1% formic acid. Retention times were 22.2 min for the glucuronide and 26.8 min for THC-COOH. After enzymatic hydrolysis of urine samples with beta-glucuronidase/arylsulfatase (37 degrees C, 5 h), THC-COOH-glucuronide was no longer detectable by LC/MS/MS in urine samples. However, the THC-COOH concentration was increased. For quantitation of THC-COOH, THC-COOH-D(3) was added to the urine samples as internal standard prior to analysis. From the difference of THC-COOH in the native urine and urine after enzymatic hydrolysis, molar concentration ratios of THC-COOH-glucuronide/THC-COOH in urine samples of cannabis users were determined and found to be between 1.3 and 4.5.
Article
The biosynthesis of cannabinoids was studied in cut sprouts of Cannabis sativa by incorporation experiments using mixtures of unlabeled glucose and [1-(13)C]glucose or [U-(13)C(6)]glucose. (13)C-labeling patterns of cannabichromenic acid and tetrahydrocannabinolic acid were analyzed by quantitative NMR spectroscopy. (13)C enrichments and coupling patterns show that the C(10)-terpenoid moiety is biosynthesized entirely or predominantly (> 98%) via the recently discovered deoxyxylulose phosphate pathway. The phenolic moiety is generated by a polyketide-type reaction sequence. The data support geranyl diphosphate and the polyketide, olivetolic acid, as specific intermediates in the biosynthesis of cannabinoids.
Article
A rapid and sensitive method for the simultaneous confirmatory analysis of three forensic most relevant cannabinoids, Delta(9)-tetrahydrocannabinol (THC), 11-hydroxy-Delta(9)-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH), by means of high-performance liquid chromatography/tandem mass spectrometry (LC/MS/MS) in human plasma was developed and fully validated. Sample clean-up was performed by automated silica-based solid-phase extraction and the separation was carried out using a PhenylHexyl column (50 x 2 mm i.d., 3 micro m) and acetonitrile-5 mM ammonium acetate gradient elution. Data were acquired with an API 3000 LC/MS/MS system equipped with a turboionspray interface and triple quadrupole mass analyzer using positive electrospray ionization and multiple reaction monitoring. Two MS/MS transitions for each substance were monitored and deuterated analogues of analytes were used as internal standards for quantitation. The limit of quantitation was 0.8 ng ml(-1) for THC, 0.8 ng ml(-1) for 11-OH-THC and 4.3 ng ml(-1) for THC-COOH and linearity with a correlation coefficient r(2) = 0.999 was achieved up to 100 ng ml(-1) for THC and 11-OH-THC and 500 ng ml(-1) for THC-COOH. The limits of detection were 0.2 ng ml(-1) for THC, 0.2 ng ml(-1) for 11-OH-THC and 1.6 ng ml(-1) for THC-COOH. The developed LC/MS/MS method was also successfully used for the determination of THC-COOH-glucuronide, the phase II metabolite of THC-COOH.
Article
A simple procedure based on a common silica gel column chromatography for the isolation of Delta9-tetrahydrocannabinolic acid A (Delta9-THCA-A) from hemp in a multi-milligram scale is presented. Further, the decarboxylation reaction of Delta9-THCA-A to the toxicologically active Delta9-tetrahydrocannabinol (Delta9-THC) at different analytical and under-smoking conditions is investigated. Maximal conversion in an optimised analytical equipment yields about 70% Delta9-THC. In the simulation of the smoking process, only about 30 % of the spiked substance could be recovered as Delta9-THC.
Article
An analytical method using solid-phase extraction (SPE) and high-performance liquid chromatography-mass spectrometry (LC-MS) has been developed and validated for the confirmation of Delta(9)-tetrahydrocannabinol (THC) in oral fluid samples. Oral fluid was extracted using Bond Elut LRC-Certify solid-phase extraction columns (10 cm(3), 300 mg) and elution performed with n-hexane/ethyl acetate. Quantitation made use of the selected ion-recording mode (SIR) using the most abundant characteristic ion [THC+H(+)], m/z 315.31 and the fragment ion, m/z 193.13 for confirmation, and m/z 318.00 for the protonated internal standard, [d(3)-THC+H(+)]. The method proved to be precise for THC, in terms of both intra-day and inter-day analyses, with coefficients of variation less than 10%, and the calculated extraction efficiencies for THC ranged from 76 to 83%. Calibration standards spiked with THC between 2 and 100 ng/mL showed a linear relationship (r(2)=0.999). The method presented was applied to the oral fluid samples taken from the volunteers during the largest music event in Portugal, named Rock in Rio-Lisboa. Oral fluid was collected from 40 persons by expectoration and with Salivette. In 55% of the samples obtained by expectorating, THC was detected with concentration ranges from 1033 to 6552 ng/mL and in 45% of cases THC was detected at concentrations between 51 and 937 ng/mL. However, using Salivette collection, 26 of the 40 cases had an undetectable THC.
Article
An ESI MS/MS library of 800 compounds has been developed and a collection of data is now available for Analyst 1.4 and higher. Compounds include forensically important drugs, such as illegal drugs, some deuterated analogues, hypnotics, amphetamines, benzodiazepines, neuroleptics, antidepressants and many others. For setting up the library of product ion spectra, 20-200 ng of the compounds have been injected either by flow injection or via a short LC-column, the precursor ions were chosen from the Q1 scan spectra, and product ion spectra were generated by CID in the collision cell using three different collision energies (20, 35 and 50 eV). Three spectra of each compound have been collected and compound names, CAS numbers, formulas and molecular weights have been added in the database, which has been generated by the Analyst software. The library can be used for compound identification during general unknown screening analysis by combination of Q1 scan techniques and subsequent MS/MS analysis in a second analytical run. Quantitative procedures for multi drug analysis using Multiple Reaction Monitoring can be established by selection of product ions and suitable collision energies from the library. For publication of the spectra, PDF-files have been generated and can be viewed on-line as supplementary data or from the website in alphabetical order: (supplementary data, should be made available via ELSEVIER-WEBSITE or via ).
Cannabis im Straßenverkehr (1.Auflage)
  • T Daldrup
  • Meininger
Daldrup T, Meininger I. Cannabis im Straßenverkehr (1.Auflage).
Blood cannabinoids II: models for the prediction of time of marijuana exposure from plasma concentrations of 9-tetrahydrocannabinol (THC) Copyright 
  • Huestis Ma Je Henningfield
  • Cone
Huestis MA, Henningfield JE, Cone EJ. Blood cannabinoids II: models for the prediction of time of marijuana exposure from plasma concentrations of 9-tetrahydrocannabinol (THC) Copyright  2007 John Wiley & Sons, Ltd. J. Mass Spectrom. 2007; 42: 354–360 DOI: 10.1002/jms J. Jung et al. and 11-nor-9-carboxy-9-tetrahydrocannabinol (THC-COOH).