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Determination of cannabinoids in hemp food products by use of headspace solid-phase microextraction and gas chromatography-mass spectrometry

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Abstract

A fully automated procedure using alkaline hydrolysis and headspace solid-phase microextraction (HS-SPME), followed by on-fiber derivatization and gas chromatographic–mass spectrometric (GC–MS) detection has been developed for determination of cannabinoids in hemp food samples. After addition of a deuterated internal standard, the sample was hydrolyzed with sodium hydroxide and submitted to direct HS-SPME. After absorption of analytes for on-fiber derivatization, the fiber was placed directly into the headspace of a second vial containing N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA), before GC–MS analysis. Linearity was good for Δ 9-tetrahydrocannabinol (THC), cannabidiol, and cannabinol; regression coefficients were greater than 0.99. Depending on the characteristics of the matrix the detection limits obtained ranged between 0.01 and 0.17 mg kg−1 and the precision between 0.4 and 11.8%. In comparison with conventional liquid–liquid extraction this automated HS-SPME–GC–MS procedure is substantially faster. It is easy to perform, solvent-free, and sample quantities are minimal, yet it maintains the same sensitivity and reproducibility. The applicability was demonstrated by analysis of 30 hemp food samples. Cannabinoids were detected in all of the samples and it was possible to differentiate between drug-type and fiber-type Cannabis sativa L. In comparison with other studies relatively low THC concentrations between 0.01 and 15.53 mg kg−1 were determined.

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... Solvent exchange is also performed prior to GC analysis, especially when derivatization is performed [7,80,87,104,121,123,133]. Extracts are reduced to dryness usually under gentle N 2 steam, causing least damage to total extracted amounts of phytocannabinoids and terpenoids than drying in rotary evaporator or in a speedvac, the latter reducing the concentrations of ∆ 9 -THC and CBG for two-thirds [66]. Reduced extracts can be dissolved in solvents (e.g., pyridine and benzene [116], CHCl 3 [5], MeCN [116], dry EtAc [55]), in a mixture of derivatization agent and solvent (e.g., toluene and BSTFA [94], pyridine and BSTFA + 1% TMCS [118], pyridine, isooctane and MSTFA [99], pyridine and BSTFA [90], pyridine and MSTFA + 1% TMCS [84]) or directly in the derivatization agent [80,99,100,133]. As silylation agents are harmful for GC injection port and column, additional evaporation to dryness is frequently performed, followed by dissolution in solvent, e.g., n-hexane [84] or MeCN [95]. ...
... Solvent exchange is also performed prior to GC analysis, especially when derivatization is performed [7,80,87,104,121,123,133]. Extracts are reduced to dryness usually under gentle N 2 steam, causing least damage to total extracted amounts of phytocannabinoids and terpenoids than drying in rotary evaporator or in a speedvac, the latter reducing the concentrations of ∆ 9 -THC and CBG for two-thirds [66]. Reduced extracts can be dissolved in solvents (e.g., pyridine and benzene [116], CHCl 3 [5], MeCN [116], dry EtAc [55]), in a mixture of derivatization agent and solvent (e.g., toluene and BSTFA [94], pyridine and BSTFA + 1% TMCS [118], pyridine, isooctane and MSTFA [99], pyridine and BSTFA [90], pyridine and MSTFA + 1% TMCS [84]) or directly in the derivatization agent [80,99,100,133]. As silylation agents are harmful for GC injection port and column, additional evaporation to dryness is frequently performed, followed by dissolution in solvent, e.g., n-hexane [84] or MeCN [95]. ...
... Extraction time depends upon matrix viscosity and lipophilicity that define the speed of diffusion of analytes from the liquid to the gas phase and, as a result, HS-SPME rate and efficiency. HS-SPME is more appropriate for simpler matrices (e.g., cannabis tea), as the extraction recoveries are proportional to the sample amount [99], while complex liquid-and protein-containing matrices cause significant matrix retention and lower recoveries, with higher LODs and lower method precision. For fatty/oily matrices, such as versatile hemp foods, alkaline hydrolysis with NaOH and Na 2 CO 3 is performed prior to HS-SPME in order to saponify the matrix lipids and reduce lipid matrix interferences [99]. ...
Article
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Cannabis is gaining increasing attention due to the high pharmacological potential and updated legislation authorizing multiple uses. The development of time- and cost-efficient analytical methods is of crucial importance for phytocannabinoid profiling. This review aims to capture the versatility of analytical methods for phytocannabinoid profiling of cannabis and cannabis-based products in the past four decades (1980–2021). The thorough overview of more than 220 scientific papers reporting different analytical techniques for phytocannabinoid profiling points out their respective advantages and drawbacks in terms of their complexity, duration, selectivity, sensitivity and robustness for their specific application, along with the most widely used sample preparation strategies. In particular, chromatographic and spectroscopic methods, are presented and discussed. Acquired knowledge of phytocannabinoid profile became extremely relevant and further enhanced chemotaxonomic classification, cultivation set-ups examination, association of medical and adverse health effects with potency and/or interplay of certain phytocannabinoids and other active constituents, quality control (QC), and stability studies, as well as development and harmonization of global quality standards. Further improvement in phytocannabinoid profiling should be focused on untargeted analysis using orthogonal analytical methods, which, joined with cheminformatics approaches for compound identification and MSLs, would lead to the identification of a multitude of new phytocannabinoids.
... The extraction methods primarily depend on the purpose and the final use of the extractants and the matrix they are contained in, i.e., as part of plant material, in a liquid preparation, or as a food additive. [89][90][91] The purpose of the extraction could range from a general liberation of total phytocannabinoids for incorporation into a cannabis product to the analytical determination of total phytocannabinoids or a specific compound of interest. [92,93] The current extraction methods summary and data are recorded in Table 2. [122] GC-MS CBG, CBN, CBD, CBC, THC Inflorescence UAE-SLE hexane Qualitative N/D [123] GC-MS 9 cannabinoids Inflorescences, fiber-type, EtOH 0.001-0.1 (LLOQ) 0.1-1 [124] GC-MS THC, CBD, CBN Food, hemp oil Hexane/IPA 1 Â 10 -4 -5 Â 10 -3 1-2 Â 10 -3 [90] Ã DAD or PDA detector unless otherwise indicated. ...
... (Pellegrini et al., 2005 [90] developed a simple LLE extraction method using hexane/isopropanol and used the (THC þ CBN)/CBD ratio to differentiate between the phenotypes of cannabis plants in different hemp food products. Lachenmeier et al., 2004 [91] have compared a fully automated procedure with alkaline hydrolysis and headspace solid-phase micro-extraction (HSSPME), followed by onfiber derivatisation with a standard LLE extraction method and shown the superiority of a fully automated method. In the study cannabinoids were extracted from hemp food products from Germany including tea, chocolate, seeds, flour, snack bars, nibbles, pastilles and shampoo approaching 100% recovery. ...
... [171] To avoid these errors, the acids must be derivatised. [172] However, as indicated in some studies, [91,117,156] complete derivatisation is difficult to achieve also causing inaccurate results as thermal degradation of cannabinoids in the injector port and column may also occur. Hazekamp et al., 2005 [160] were comparing different chromatographic techniques on a model of D9-THC, they noted D8-THC and CBN were present on GC chromatograms in significant amounts in contrast to HPLC, NMR, and TLC techniques, both D8-THC and CBN are known degradation products of D9-THC. ...
Article
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The legalization of the cultivation of low Δ9-tetrahydrocannabinol (Δ9-THC) and high cannabidiol (CBD) Cannabis Sativa plants is gaining momentum around the world due to increasing demand for CBD-containing products. In many countries where CBD oils, extracts and CBD-infused foods and beverages are being sold in health shops and supermarkets, appropriate testing of these products is a legal requirement. Normally this involves determining the total Δ9-THC and CBD and their precursor tetrahydrocannabinolic acids (THCA) and cannabidiolic acid (CBDA). As our knowledge of the other relevant cannabinoids expands, it is likely so too will the demand for them as additives in many consumer products ensuring a necessity for quantification methods and protocols for their identification. This paper discusses therapeutically relevant cannabinoids found in Cannabis plant, the applicability and efficiency of existing extraction and analytical techniques as well as the legal requirements for these analyses.
... Many studies have described analytical methods for determining cannabinoid levels in hemp seeds. These methods include microwave, sonication, soxhlet [5], liquid-liquid extraction (LLE) [13], solid-phase extraction (SPE) [16], and solid-phase microextraction (SPME) [17] extraction and without derivatization method [7,13,18-20], or derivatization with N-methyl-Ntrimethylsilyltrifluoroacetamide (MSTFA) [17,21], or tetrabutylammonium hydroxide (TBAH) [22] were used. ...
... Many studies have described analytical methods for determining cannabinoid levels in hemp seeds. These methods include microwave, sonication, soxhlet [5], liquid-liquid extraction (LLE) [13], solid-phase extraction (SPE) [16], and solid-phase microextraction (SPME) [17] extraction and without derivatization method [7,13,18-20], or derivatization with N-methyl-Ntrimethylsilyltrifluoroacetamide (MSTFA) [17,21], or tetrabutylammonium hydroxide (TBAH) [22] were used. ...
... mg/g in Chinese seeds [6], 0.95-1.08 mg/g in Swiss seeds [17], 0.02-1.49 mg/g in Taiwanese seeds from [6], and 8À12 mg/g in Hungarian seeds [15]. ...
Article
Hemp seeds and hempseed oil are marketed on- and off-line as health foods and cosmetics and have been reported to have high nutrient contents. However, because of the various side effects of cannabinoids, especially △9-tetrahydrocannabinol (THC), many countries regulate upper limits for THC in products, which creates the need for analytical techniques capable of measuring THC, cannabidiol (CBD), and cannabinol (CBN) levels in commercial hemp seeds and hempseed oil. In the present study, hemp seed and hempseed oil extracts obtained by methanol extraction, were analyzed by gas chromatography-mass spectrometry (GC/MS). Validation of the technique used was performed using calibration curves and by determining LODs, LOQs, specificities, selectivities, and intra- and inter-day precision and accuracies. In addition, matrix effects, process efficiencies, recoveries, and sample stabilities were investigated. In hemp seeds, as determined using the fully optimized method THC concentrations ranged from 0.06 to 5.91 μg/g, CBD concentrations from 0.32 to 25.55 μg/g, and CBN concentrations from 0.01 to 1.50 μg/g; CBN/THC ratios ranged from 0.1 to 1.60, and CBD/THC ratios from 0.11 to 62.56. Furthermore, the (THC + CBN)/CBD ratio of most hemp seed samples was less than one. In hempseed oil, THC concentrations ranged from 0.3 to 19.73 μg/mL, CBD concentrations from 6.66 to 63.40 μg/mL, CBN concentrations from 0.11 to 2.31 μg/mL, CBN/THC ratios from 0.12 to 0.42, and CBD/THC ratios from 3.21 to 22.50. Furthermore, (THC + CBN)/CBD ratios in all hempseed oil samples were less than one. The optimized methanol extraction-GC/MS technique was found to be satisfactory for determining THC, CBD, and CBN concentrations in hemp seeds and hempseed oil.
... Zoller, Rhyn and Zimmerli (2000) developed a HPLC-UV/DAD method to quantify Δ 9 -THC and THCA-A in hemp seed, hemp seed oil, biscuits, tea and herbal hemp, but no samples were analysed. In 2004 Lachenmeier, Kroener, Musshoff andMadea (2004) determined Δ 9 -THC, CBD and cannabinol (CBN) levels by GC/ MS in 29 samples of food containing hemp, including tea, chocolate, seed, flour, various snack, soft drink and beer. An interesting survey by Holler, Bosy, Dunkley, Levine, Past and Jacobs (2008) reported the content of Δ 9 -THC in 79 commercial food and beverages containing hemp, such as tea, beer, chips, cereals, flours, pretzels and vodka. ...
... Zoller, Rhyn and Zimmerli (2000) developed a HPLC-UV/DAD method to quantify Δ 9 -THC and THCA-A in hemp seed, hemp seed oil, biscuits, tea and herbal hemp, but no samples were analysed. In 2004 Lachenmeier, Kroener, Musshoff andMadea (2004) determined Δ 9 -THC, CBD and cannabinol (CBN) levels by GC/ MS in 29 samples of food containing hemp, including tea, chocolate, seed, flour, various snack, soft drink and beer. An interesting survey by Holler, Bosy, Dunkley, Levine, Past and Jacobs (2008) reported the content of Δ 9 -THC in 79 commercial food and beverages containing hemp, such as tea, beer, chips, cereals, flours, pretzels and vodka. ...
Article
In 2016, the European Commission recommended the Member States to monitor the content of Δ⁹-tetrahydrocannabinol and other cannabinoids in food and feed derived from hemp and in food of animal origin for possible transfer from feed. Thus, the Italian Ministry of Health implemented a monitoring plan. To this aim, nine cannabinoids in beverages and food for human consumption and in feed were determined. The method applied, based on rapid clean-up and LC-MS/MS determination, was previously developed and in-house validated, evaluating the analytical performance in the concentration ranges 2-50 µg/L for beverages, 0.020-0.500 mg/kg for food and 0.100-10.0 mg/kg for feed. Then, it was applied to determine the cannabinoids in 78 food, 16 beverage and 6 feed samples, collected from the Italian market since 2017. The results are herein reported, for evaluation of both product characteristics and compliance to national maximum limits. Some study cases are also described.
... As a rule, solid-fluid or liquid-liquid extraction was used to prepare the samples with various amounts of such extraction solvents as mixtures of methanol with three chloromethane, ethyl acetate with isopropanol, mixtures of methanol or acetonitrile with water, 2-propanol, or 95% ethanol. Chromatographic methods, mainly HPLC-UV [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16], and more rarely GC-MS [17,18] or LC-MS/MS [16,19] techniques, were chosen as separation and quantification methods. ...
... As a rule, solid-fluid or liquid-liquid extraction was used to prepare the samples with various amounts of such extraction solvents as mixtures of methanol with three chloromethane, ethyl acetate with isopropanol, mixtures of methanol or acetonitrile with water, 2-propanol, or 95% ethanol. Chromatographic methods, mainly HPLC-UV [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16], and more rarely GC-MS [17][18] or LC-MS/MS [16,19] techniques, were chosen as separation and quantification methods. ...
Article
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Currently, the medical use of food supplements containing Cannabis sativa has attracted the interest of consumers, as well as the medical and scientific community. With the increasing consumption of these products, there is also a risk of their abuse or discrepancy between the actual and declared contents of active substances by the manufacturer in these products. Thus, the development and elaboration of analytical procedures for determination of appropriate phytocannabinoids seems to be important. This work focuses on the development of a simple, fast and environmentally friendly liquid-liquid extraction method combined with fat freezing from an oil sample to isolate two phytocannabinoids: cannabidiol (CBD) and cannabidiolic acid (CBDA). The extraction method was optimized considering efficacy and repeatability of extraction, as well as minimalizing use of organic reagents and sample amount. Under the optimized conditions, extraction recovery for CBD was 97.3–109% and for CBDA was 69.1–69.5% with precision (RSD, %) 5.0–8.4 and 7.1–10.6, respectively. The evaluated main analytical parameters of the developed high pressure liquid chromatography with diode array detector (HPLC-DAD) method for both studied cannabinoids are satisfactory. The usability of the developed method was checked by analysis of real samples of a food supplement–hemp oil enriched with CBD.
... Selbst in dieser Umfrage von 2019 wurde die Verwendungshäufigkeit von CBD als sehr ge-ring beschrieben und auf die Aktualität vieler Erstanwender hingewiesen [3]. Frühere Veröffentlichungen unserer eigenen Arbeitsgruppe [4][5][6][7] wurden fälschlicherweise (z. B. in [8,9]) als Beleg für eine Verfügbarkeit von Hanfextrakten vor 1997 angeführt. ...
... Es wird hierzu ausdrücklich darauf hingewiesen, dass die Probenahmen bzw. Marktübersichten der genannten Veröffentlichungen [4][5][6][7] in den Jahren 2002/2003 erfolgten und die dort genannten Produkte keinerlei Extrakte beinhalteten. Neben CBD und Hanfextrakten ist auch die Verwendung von Hanfblüten in der Regel als "neuartig" einzustufen (z. ...
Article
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Zusammenfassung Im Novel-Food-Katalog der Europäischen Kommission sind Cannabidiol (CBD) und Hanfextraktprodukte als neuartig eingestuft und benötigen somit vor dem ersten Inverkehrbringen eine Zulassung. Vonseiten der Lebensmittelunternehmen wird aber eine Vielzahl derartiger Produkte ohne Zulassung in den Verkehr gebracht und von der amtlichen Überwachung beanstandet. Sieben Gerichtsurteile haben mittlerweile einheitlich die Einstufung als Novel Food untermauert und die amtlichen Maßnahmen bestätigt. Es ist zu hoffen, dass sich die Lebensmittelunternehmen der Problematik bewusst werden und ihrer Sorgfaltspflicht nachkommen, indem sie die Sicherheit der Produkte im Rahmen eines Zulassungsverfahrens belegen. Eine große Rechtsunsicherheit besteht allerdings weiterhin vor dem Hintergrund einer möglichen Betäubungsmitteleinstufung von Hanflebensmitteln, insbesondere solchen aus Blättern (beispielsweise als Tee). Summary In the Novel Food catalogue of the European Commission, cannabidiol (CBD) and hemp extract products are classified as novel and therefore require a marketing authorisation before being placed on the market for the first time. Contrary to this, a large number of such products has been placed on the market by food companies without approval and has been objected by the official control authorities. Seven court rulings meanwhile have uniformly supported the classification as Novel Food and have confirmed the official measures. It is to be hoped that the food business operators will become aware of the problem and fulfil their duty of care by proving the safety of products within the framework of approval procedures. However, a great deal of legal uncertainty still exists within the framework of a possible narcotic classification of hemp foods, especially those made from leaves (e.g. as tea).
... Abhängig vom THC-Gehalt kann zwischen Drogenhanf und Faserhanf unterschieden werden. Die Phänotypen von Cannabis sativa werden durch das Verhältnis (THC+CBN)/CBD charakterisiert (Drogenhanf oder Marihuana > 1; Faserhanf < 1) [15][16][17][18]. Die größten Drüsenhaare werden an weiblichen Hanfpflanzen in den Blütenregionen und hier besonders auf den Blättern und Samenhüllblättern vorgefunden. ...
... Als Alternative zu diesen etablierten Probenvorbereitungsmethoden kann die Headspace(HS)-Festphasenmikroextraktion (Solid-Phase Microextraction, SPME) eingesetzt werden. Trotz der nur geringen Flüchtigkeit der Cannabinoide und der Möglichkeit einer Phenolatbildung im alkalischen Aufschlussmedium lässt sich diese Stoffgruppe mittels SPME reproduzierbar aus dem Headspace extrahieren, da die lipophilen Verbindungen vergleichsweise hohe Oktanol-Wasser-Verteilungskoeffizienten aufweisen und damit eine hohe Affinität zur unpolaren Polydimethylsiloxan(PDMS)-SPME-Faser besitzen[108][109][110]. Eine vollständig automatisierte HS-SPME-Methode zur Bestimmung von THC, CBD und CBN in allen Arten von Hanflebensmitteln wurde von Lachenmeier et al.[18] entwickelt. Die Lebensmittelproben werden nach Zugabe eines deuterierten internen Standards mit Natriumhydroxid hydrolysiert und direkt mit HS-SPME/GC/MS vermessen. ...
Article
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Zusammenfassung Seit dem 1996 aufgehobenen Anbauverbot für Pflanzen der Spezies Cannabis sativa L. (sog. Faserhanf) mit geringem Gehalt des psychoaktiven Inhaltsstoffs Δ9-Tetrahydrocannabinol (THC) wird eine Vielzahl daraus hergestellter Lebensmittel angeboten. Als Beurteilungshilfe für die amtliche Lebensmittelüberwachung werden in dieser Übersichtsarbeit alle Aspekte von Hanf als Lebensmittel diskutiert, eine Einführung in die Botanik der Hanfpflanze gegeben und die aktuelle Gesetzeslage in Deutschland und der Europäischen Union dargestellt. Forensisch-toxikologische Aspekte insbesondere hinsichtlich des Einflusses von Hanflebensmitteln auf Drogentests werden beschrieben und eine Übersicht über die analytischen Möglichkeiten zur Absicherung der THC-Richtwerte gegeben. Abschließend werden Vorschläge für die lebensmittelchemische und rechtliche Beurteilung von Hanflebensmitteln gemacht. Neue Aspekte in diesem Update betreffen insbesondere sog. Cannabidiol (CBD)-Öle und deren Novel-Food-Status. Seit 1998 wurde ein Rückgang der THC-Konzentrationen für mehrere Produktgruppen beobachtet. Die von der EU vorgeschriebene Verwendung von zertifiziertem Hanfsamen und die verstärkte Kontrolle der Hersteller haben offensichtlich zu einem deutlichen Rückgang der THC-Konzentrationen in Hanflebensmitteln geführt. Der maximale THC-Gehalt in derzeit verfügbaren traditionellen Hanflebensmitteln ist zehn- bis hundertfach niedriger als in den Studien der 1990er-Jahre. Es ist zu beachten, dass frühere GC-Studien immer die Summe von THC und THC-Säuren bestimmten. In den letzten Jahren liefern LC-MS-Methoden Informationen über den spezifischen Gehalt an THC in Hanfprodukten. Dennoch wurden seitdem immer noch Lebensmittel mit inakzeptablen THC-Gehalten vorgefunden, was zu einer ganzen Reihe von öffentlichen Warnungen im EU-Schnellwarnsystem für Lebens- und Futtermittel (RASFF) führte. Daher ist eine kontinuierliche Qualitätskontrolle erforderlich, um den THC-Wert niedrig zu halten. Dazu gehört sowohl die Verwendung von Sorten mit niedrigem THC-Gehalt als auch die richtige Saatgutreinigung. Jüngstes Interesse gilt dem CBD, das wegen seiner vermeintlich günstigen gesundheitlichen Eigenschaften vermarktet wird. Während natürliche Gehalte in den oben genannten Lebensmitteln toleriert werden, werden reine CBD-Extraktprodukte entweder als Arzneimittel oder als sogenanntes Novel Food behandelt, die beide vor dem Inverkehrbringen zugelassen werden müssen. Nicht-traditionelle Hanf-Extraktprodukte weisen zudem häufig so hohe THC-Konzentrationen auf, dass die Produkte als gesundheitsschädlich beurteilt werden müssen. Summary In 1996, the prohibition of the cultivation of plants of the species Cannabis sativa L. (so-called fibre hemp) with minor content of the psychoactive Δ9-tetrahydrocannabinol (THC) was lifted. Nowadays, a wide variety of hemp food products is offered on the market. As a help for evaluation of such products, this review article provides the official food control with information on all aspects of hemp as foodstuff. An introduction to the botany of the hemp plant and the current law situation in Germany and the European Union is presented. In particular, the forensic-toxicological aspects regarding the influence of hemp food on drug tests are described. Furthermore, an overview of the analytic techniques used to verify compliance with the guidance values is given. Finally, suggestions for the food regulatory and food chemical evaluation of hemp food products are made. New aspects in this update concern so-called cannabidiol (CBD) oils and their novel food status. Since 1998, a decrease in the THC concentrations for several product groups has been observed. The prescribed use of certified hemp seed by the EU and the increase of controls on manufactur-ers have obviously led to a significant decline of THC concentrations in hemp food products. The maximum THC content in currently purchasable traditional hemp food products is ten- to a hundred-fold lower than those found in the studies of the 1990s. It is of note that earlier GC studies always consider the sum of THC and THC-acids. In the last years, LC-MS methods provide information about the isolated content of THC in hemp products. Nevertheless, food products with inacceptable THC contents were still observed since then, which led to a series of public warnings in the EU Rapid Alert System for Food and Feed (RASFF). Therefore, ongoing quality control is needed to maintain low THC levels. This includes both the use of low THC varieties and proper seed cleaning. Recent interest surrounds CBD, which is purported for various health properties. While natural contents in the above-mentioned food products are tolerated, pure CBD extract products are either treated as medicines or as so-called novel food, which both need to be approved before being placed on the market. In addition, nontraditional hemp extract products often exhibited extreme THC concentrations, which must be evaluated as hazardous to health.
... Bosy et al. [27] reported 6 hemp seed oil products from USA and Canada with a THC content ranging from 11.5 to 117.5 μg/g. Lachenmeier et al. [28] reported 30 hemp food products from Germany including tea, chocolate, seeds, flour, snack bars, nibbles, pastilles, drinks, oil, and shampoo with a THC content ranging from 0.01 to 15.53 μg/ g, a CBD content ranging from 0.03 to 62.59 μg/g, and a CBN content ranging from 0.01 to 4.19 μg/g. Stolker et al. [23] reported 2 cannabis products from the Netherlands with a THC content of 1400 and 3100 μg/g and a CBD content of 20 and 10 μg/g, respectively. ...
... [39] Interestingly, the results of investigation of hemp products were not consistent. Even if the cannabinoids content in hemp products was extremely low, some studies [23,24,40] have demonstrated higher than 1:1 of CBD-to-THC ratios with a majority; whereas some studies, [25,28] which were similar to our findings, reported lower than 1:1 of CBD-to-THC ratios with a majority. Although it is hard to distinguish the strains of the prepared concentrated powder products in the present study, a study evidenced that industrial hemp is genetically more similar to C. indica type, whereas marijuana is more similar to C. sativa strains. ...
Article
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Hemp nuts are mature cannabis seeds obtained after hulling and stir-frying that are commonly used in traditional Chinese medicine for treating functional constipation. In this work, we screened hemp nut products, classified them, and verified the legality of consuming them. A total of 18 products were purchased from Taiwan, China and Canada. Validated high-performance liquid chromatography with tandem mass spectrometry methods were developed for analyzing the cannabinoid (i.e., Δ(9) -tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol) content of the products and the concentration of urinary 11-nor-9-carboxy-THC. Chemometric techniques, namely hierarchical clustering analysis (HCA) and principal component analysis (PCA), were applied for rapidly classifying 11 concentrated powder products in Taiwan. A pilot human study comprising single and multiple administrations of a product with 1.5 µg/g of THC was conducted to examine the urinary 11-nor-9-carboxy-THC concentration. Through optimization of 3(2) full factorial design, using 60% isopropanol as the extraction solvent exhibited the highest yield of cannabinoids and was applied as the optimal condition in further analysis. The results of HCA and PCA on quality evaluation were in well agreement; however, the tested products possessed distinct CBD-to-THC ratios which ranged widely from 0.1:1 to 46.8:1. Particularly, the products with CBD-to-THC ratios higher than 1:1 were the majority in Taiwan. Our data suggested that all the tested hemp nut products met the Taiwan restriction criteria of 10 µg/g of THC. We propose a usual consumption amount of hemp nut products in Taiwan would unlikely to violate the cut-off point of 15 ng/mL of urinary 11-nor-9-carboxy-THC. This article is protected by copyright. All rights reserved.
... Beside oral fluid and hair samples for which plenty of SPME applications have been reported; SPME has also been applied for the determination of cannabinoids in other relevant matrices of forensic interest such as food, water and cannabis samples. In a study performed by Lachenmeier et al. [30], cannabinoids (Δ 9 -THC, CBD and CBN) have been extracted from hemp food products by HS-SPME using 100 μm PDMS followed by on-fiber derivatization with MSTFA. The whole sample preparation procedure was fully automated where alkaline hydrolysis, HS-SPME and on-fiber derivatization were performed using a programmable autosampler. ...
... The authors compared HS-SPME procedure with LLE using 9:1 n-hexane: ethyl acetate. Comparative chromatograms of HS-SPME and LLE clearly proved the superiority of HS-SPME over LLE; since several interfering peaks at the retention times of analytes were observed in chromatogram of LLE which were absent in HS-SPME chromatogram [30]. ...
Article
Analysis of cannabinoids in biological and other matrices is of paramount importance in forensic toxicology, since they are the most widely abused drugs over the globe. Generally, extraction of cannabinoids from biological matrices is achieved by liquid-liquid extraction (LLE) and solid-phase extraction (SPE). However, great attention has now been paid towards modern microextraction techniques in order to improve the quality and sensitivity of analytical methods. Microextraction techniques are environmentally benign, rapid, sensitive, inexpensive, offer high extraction efficiency and enrichment factors. This review provides an overview of microextraction techniques applied for the determination of cannabinoids and their metabolites in oral fluid, hair, urine and other matrices of forensic importance. After a complete revision of microextraction techniques for cannabinoids, they have been classified into two categories: (i) solid based e.g. solid-phase microextraction (SPME) etc., and (ii) solvent based e.g. dispersive liquid-liquid microextraction (DLLME).
... Dados do Século XXI mostram que em análises do óleo das sementes nos Estados Unidos 117 ppm (L/mL) era  9 -THC, na Alemanha 214 ppm e na Suíça 3568 ppm [23] . O elevado teor suíço deve-se ao processamento de sementes de plantas com alto teor de canabinóides [24] . Neste país todas as variedades do gênero podem ser cultivadas legalmente, embora o limite de  9 -THC nos alimentos seja regulamentado [25] . ...
... Ancak kenevirin kullanıldığı gıdalarda uyuşturucu etkisi ilk paragrafta bahsettiğimiz gibi merak edilmekte bu alanda gıdalarda uyuşturucu özellik gösteren temel bileşenin analizleri çok rahat bir şekilde yapılmaktadır. Çalışmalar endüstriyel kenevirde uyuşturucu bileşeninin tespit edilemediği veya çok düşük konsantrasyonlarda bulunduğu ve bu miktarın uyuşturucu özellik göstermediğini ispatlamıştır (13,14). ...
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K enevir birçok farklı alanda kullanılan önemli bir bitkidir. Hem erkek hem de dişi yapısı olan, yıllık dioik bir türdür. Hint keneviri (Cannabis indica) ve endüstriyel kenevir olarak adlandırılan (Cannabis sativa L.) iki türü öne çıkmaktadır. Narko-tik maddeler için üretilen ve dişi çiçek salkımlarının reçinesi olan tür indica'dır. Sativa ise güçlü lifler için üretilen ve çok az reçine oluşturan en yaygın türdür. Günümüzde yaşanan kuraklıklar ve savaşlar nedeni ile buğday stoklarında ve üretiminde sıkıntılar baş göstermeye başlamıştır. Bu nedenle buğday unu yerine unlu mamullerin yapımında kullanılmak üzere alternatif ürünlerin arayışına gidilmektedir. Bu alan da ihtiyacı gidermek için kenevirin kullanımı da araştırılmaktadır. Kenevir denilince ilk akla gelen uyuşturucu olarak tüketil-mesi nedeni ile kuşku ile bakılmaktadır. "Acaba böyle bir etkiye sahip olan kenevir ile biz uyuşturucu bağımlısı mı yapılmak isteniyoruz?" sorusu zihinleri meşgul etmektedir. Bu soru doğal olarak herkesin merak ettiği bir konudur. Fakat kenevirin uyuşturucu olarak kullanıldığı tür, tıbbi kenevir olarak bilinen türüdür. Kenevirin, endüstriyel kenevir olarak adlandırılan türü birçok farklı sanayi alanında da kullanıldığı için endüstriyel kenevir olarak isimlendirilmekte ve unlu mamullerin yapımında bu tür kullanılmaktadır. Bu ne-denle endüstriyel kenevirden elde edilen un ile üretilen mamullerde uyuşturucu etkisinin oluşmadığını bilimsel çalışmaların sonuçları ve gönül rahatlığı ile söyleyebiliriz. Haşhaş da uyuşturucu olarak etkisi bilinen ama aynı zamanda birçok unlu mamullerde kullanılan bir bitki türü olduğu için bu yönü ile kenevirde haşhaş gibi kullanılabilmektedir. Böylece gelecekte ön görülen gıda stoklarındaki azalmada kenevirin alternatif bir ürün olabileceği görülmektedir. Bu alanda özellikle ekmek, kurabiye ve pasta gibi unlu mamullerde kullanıl-ması hem ekonomik hem de sosyal yönden topluma önemli derecede katkı sağlayacaktır (1). Kenevir, uyuşturucu özelliğinden dolayı uzun yıllar yasaklanmasına rağmen dünya genelinde birçok ülkede son zamanlarda yapılan düzenlemeler ile üretimi teşvik edilmektedir. Ülkemizde de geçmişte yapılan yasaklamalar yerini kontrollü üretime bırakmış ve 2016 yılında yapılan düzenleme ile üretimi daha da teşvik edilmiştir. Bu amaçla kenevir ile ilgili bir yönetmelik hazırlanmıştır. Bu çıkarılan yönetmelik ile kenevirin uyuşturucu olarak kullanılması engellenmekte ve endüstriyel kenevirin ise belirli alanlarda kullanılması için üretimi teşvik edilmekte, üretimin yapılacağı usul ve esaslar temel hatları ile ortaya konmaktadır (2). Kenevirin gıda kaynağı olarak kullanımı yüzyıllar öncesine dayanmaktadır. Özellikle Avrupa'da besleyici gıda olarak kullanılmaktadır. Gıda sektöründe doğrudan ve dolaylı
... Many articles related to CBD and other cannabinoids determination in plants [8], oils [9], or other matrices, such as biological fluids, hair, or food products can be found in the analytical literature based on the use of different sample preparation strategies [10] and chromatographic techniques [11]. Gas chromatography coupled to simple mass spectrometry (GC-MS) was used for the analysis of hair [12][13][14][15][16], oral fluid [17,18], and hemp food [19] samples. Gas chromatography with tandem mass spectrometry (GC-MS/MS) was also used for hair sample analysis [20][21][22][23]. ...
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Cannabidiol is a phytocannabinoid with proven pharmacological properties that is also used in the cosmetic industry for its sebostatic and antioxidant activities, being considered a new anti-aging ally. An analytical method is proposed for the determination of CBD in cosmetic products by liquid chromatography with tandem mass spectrometry, after leaching the CBD from the cosmetic matrix with ethanol. Low instrumental limits of detection (0.22 ng mL−1) and quantification (0.74 ng mL−1) allow the determination of CBD at trace levels without needing preconcentration, whereas the wide linearity of the method allows the determination of CBD in more concentrated samples without high dilution. The method was successfully applied to the analysis of six cosmetic products and a raw material. The proposed method is suitable for the quality control of cosmetic products containing CBD, being able to quickly and easily determine this compound, ensuring that its concentration in the finished product is the desired one.
... Latin America is the world leader in the promotion and approval of regulations enabling the medicinal use of C. sativa (Aguilar et al. 2018). Lachenmeier et al. (2004) recall that in the EU, the allowed Δ 9 -THC content was set at 0.5% in 1984. In France, Hungary, and former Soviet republics, limits on Δ 9 -THC content were gradually made stricter due to the growing drug abuse, reaching 0.3% in 2001 (Lachenmeier and Walch 2005). ...
Article
Influence of agroclimatic conditions on active substances content in hemp cultivated in the South-East Baltic region Abstract: Due to legal regulations prohibiting cultivation of cannabis (Cannabis indica Lam.) in many countries, industrial hemp (Cannabis sativa L.) remains the main source of active substances with potential application in the pharmaceutical industry. To assess the possibility of using the varieties of industrial hemp for this goal, and their adaptation to the habitat conditions and the agricultural technology appropriate for them, we investigated three monoecious varieties of hemp (‘Futura 75,’ ‘KC Dora’ and ‘Tygra’) at different sowing densities (60 germinating seeds·1 m² or 180 germinating seeds·1 m²) and nitrogen fertilization levels (0, 30, 60, 90 kg·ha⁻¹). In none of the tested hemp varieties registered as fibrous did the concentration of Δ⁹-THC exceed 0.2%, satisfying the requirements of European legislation for industrial hemp varieties. The tested varieties did not differ significantly in the cannabidiol (CBD) and Δ⁹- tetrahydrocannabinol (Δ⁹ -THC) content in the dry matter of inflorescences. Agronomic factors such as sowing density or nitrogen fertilization did not modify the content of CBD and Δ⁹-THC. This result is very helpful for farmers, because it allows them to select hemp varieties of dual-purpose production (stems and inflorescences or stems and seeds) adapted to the South-East Baltic environment.
... The German Federal Institute for Risk Assessment (BfR) even suggests a slightly higher amount of 2 g / 200 mL infusion (10 g/L) and suggests that the actual acute consumption quantities for herbal tea as an analogon for hemp tea are in the order of 1.3 litres (P95) 19 . Regarding the question of carry-over of THC into the infusion, the BfR has recently reviewed the evidence including the study cited by EIHA 20 and another study by our group 21 . The BfR concluded: "The BfR is of the opinion that the assumption of 100% carryover is justified, as experimental data on the carryover point to high fluctuations" 19 . ...
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An interesting and valuable discussion has arisen from our recent article (Lachenmeier et al., 2020) and we are pleased to have the opportunity to expand on the various points we made. Equally important, we wish to correct several important misunderstandings that were made by Kruse and Beitzke (2020) on behalf of the European Industrial Hemp Association (EIHA) that possibly contributed to their concerns about the validity of our data, toxicological assessment and conclusions regarding regulatory status of cannabidiol (CBD) products. First and foremost, our study did only assess the risk of psychotropic Δ ⁹ -tetrahydrocannabinol (THC) without inclusion of non-psychotropic Δ ⁹ -tetrahydrocannabinolic acid (THCA). Secondly, as this article will discuss in more detail, there is ample evidence for side effects of CBD products, not only in paediatric patients, but also in adult users of over-the-counter CBD products (including inadvertent “high” effects). Thirdly, the exposure and risk assessment was conducted using up-to-date guidelines according to the European Food Safety Authority (EFSA) and the German Federal Institute for Risk Assessment (BfR). And finally, the current legal situation in the European Union, without approval of any hemp extract-containing product according to the Novel Food regulation, actually allows blanket statements that all such products are illegal on the market, and this indeed would imply a general ban on the use and marketing of such products as food or food ingredients until such an approval has been granted. We hope that this reassures the F1000Research readership regarding the validity of our results and conclusions. We are pleased, though, that the EIHA has acknowledged the fact that there are non-compliant CBD products available, but according to our data these are a substantial fraction of the market.
... The hemp fibre had a considerable history in terms of providing high tensile strength, especially in the use for roping, and in being part of a large productive system [7]. ...
Article
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As result of the increasing environmental awareness, the interest of bio-composite materials is growing rapidly in the last years in order to use them in various engineering application fields. For some of these applications, drilling is often required to facility the assembly of the parts and to make final products. However, drilling can induce a number of problems such as delamination, pull-out and strength reduction depending on the used cutting tools and cutting process parameters that can negatively affect the final product properties. Therefore, in order to reduce these problems on hemp/epoxy composites the aim of this study is to evaluate the effect of both the drilling parameters and tools on the drilling forces and delamination factors.
... Mit einer Verringerung der THC-Gehalte in Hanflebensmitteln durch sorgfältigere Behandlung zur Vermeidung von Kontaminationen ging eine Verringerung der THC-Metabolit-Gehalte im Urin der Konsumenten einher. Aus einer Studie von 2003 mit sechs Testpersonen konnten nach einem Konsum von sechs Tassen Hanftee (je 0,2 L) innerhalb von zwei Stunden mittels einer immunchemischen Standardscreening-Methode keine THC-Metabolite im Urin nachgewiesen werden [48]. Bestätigt wurde dies auch durch Ergebnisse anderer Arbeitsgruppen, wonach aus einem übermäßigen Konsum aktuell im Handel erhältlicher hanfsamenbasierter Lebensmittel keine positiven Urinanalysen resultieren [5,[43][44][45][46][47][49][50][51]. ...
Article
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Kurzfassung: Hanfhaltige Lebensmittel liegen voll im Trend. Großer Popularität erfreuen sich derzeit neuartige Nahrungsergänzungsmittel mit Cannabisextrakt, für die vor allem mit dem nicht-psychotropen Cannabinoid Cannabidiol (CBD) und dessen vermeintlich positiven Wir-kungen auf die Gesundheit geworben wird. Seit dem Aufkommen dieser CBD-Produkte werden durch die Lebensmittelüberwachungsbehörden extrem überhöhte ∆ 9-Tetrahydrocannabinol (THC)-Gehalte berichtet, die von klassischen Hanflebensmitteln nie erreicht wurden. Die An-wesenheit von THC in hanfbasierten Lebensmitteln hat neben der Problematik möglicher psy-chotroper Effekte auch Bedenken aufgeworfen, dass bei Drogentests positive Ergebnisse erhal-ten werden. Cannabis-positive Ergebnisse bei Blut-bzw. Urinuntersuchungen wurden bislang als ein Hinweis auf die Aufnahme von Cannabis, in der Form von Haschisch oder Marihuana, interpretiert und können daher unangenehme Folgen für den Betroffenen haben. In ersten Stu-dien nach Aufkommen der Hanflebensmittel wurden positive Ergebnisse bei forensisch-toxi-kologischen Tests auf Haschisch oder Marihuana nach dem Konsum von Hanfsamenöl und anderen Hanfsamenlebensmitteln beschrieben. Die verfügbare Literatur zeigt jedoch eine große Spanne, welche oralen THC-Gehalte gesichert zu einem positiven Drogentest führen würden. In der Regel ist ein negativer Befund bei einer Dosis von weniger als 0,1 mg/Tag wahrschein-lich. In eigenen Untersuchungen von CBD-Produkten (n=28) wird bei bestimmungsgemäßer Verwendung bei 43% der Produkte bereits eine THC-Dosis von mehr als 1 mg aufgenommen. Durch das Aufkommen zumeist illegal vertriebener CBD-Produkte mit teilweise sehr hohen THC-Gehalten wurde das Risiko eines positiven Drogentests somit wieder erhöht.
... For example, the German target value to exclude any risk of THC in foods would be 150 µg/kg (0.000015%) [22][23][24], and only such a level would also exclude any risk of a positive drug test with certainty. Sensitive methods, such as combinations of gas or liquid chromatography with tandem mass spectrometry (MS/MS), are necessary for adequate quality control [13,25]. Methods with unspecific and insensitive detectors, such as flame-ionization detection (FID) are typically inadequate. ...
Article
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Cannabidiol (CBD) is a non-psychoactive cannabinoid, widely marketed to athletes for claimed effects such as decreased anxiety, fear memory extinction, anti-inflammatory properties, relief of pain and for post-exercise recovery. The World Anti-Doping Agency (WADA) has excluded CBD from its list of prohibited substances. Nevertheless, caution is currently advised for athletes intending to use the compound—except CBD, all other cannabinoids are still on the prohibited list. CBD products, specifically non-medicinal, so-called full-spectrum cannabis extracts, may contain significant levels of these substances, but also contaminations of tetrahydrocannabinol (THC) (>2.5 mg/day in >30% of products on the German market) potentially leading to positive doping tests. Labelled claims about CBD content and absence of THC are often false and misleading. Contaminations with the psychoactive THC can result in adverse effects on cognition and, in general, the safety profile of CBD with respect to its toxicity is a controversial topic of discussion. For these reasons, we would currently advise against the use of over-the-counter CBD products, especially those from dubious internet sources without quality control.
... The hemp fibre had an important history in terms of providing high tensile strength, especially in the use for roping, and in being part of a large productive system [27]. After a few decades of oblivion, due to drug production-related issues, the availability of varieties with low tetrahydro-cannabinoids (THCs) content allowed the hemp production to resume [28]. Therefore, it is necessary to raise up the profile of the use of this fibre towards more engineered components, also considering that the hemp plant is available as removable resource, can easily be grown around the world and has the ability to extract heavy metals from the soil makes. ...
Article
Lightweight composite materials are frequently used for transportation, or the interiors of furniture and boxes. Wear of the surfaces of these materials is a potential health risk affecting the respiratory system or skin. The latter can frequently occur due to human touch of uncovered synthetic fibres after wear causing dermatitis, or inflammation of the skin. Therefore, composite materials made of natural fibres as reinforcement are an interesting alternative to synthetic fibres, because they are usually less dangerous to human health. Therefore, the goal of this research is to highlight the wear resistance of hemp fibres and compare it with glass and carbon fibre composites. In this work, hemp, glass and carbon fibres in form of woven fabrics were impregnated with epoxy resin through vacuum infusion process. In order to compare the tribological behaviour of the manufactured composites, a detailed experimental campaign, including tribological tests, microgeometrical measurements and indentation tests, was carried out. The tribological behaviour was studied through the pin-on-disk tests under different conditions that mainly differ in the applied load and both the composite and the single un-impregnated fabrics were tested. The results demonstrate good wear behaviour of the laminates reinforced by hemp fibres emphasising a better wear resistance at prolonged time and under high load conditions.
... Jain and Singh (2016) [273] have made and excellent and extensive review on these miniaturized procedures applied to cannabinoids determination. Among the most common techniques are solid-phase microextraction (SPME) [316][317][318][319][320][321][322] solid-phase dynamic extraction (SPDE) [323,324], microextraction by packed sorbents (MEPS) [325,326], and dispersive liquid-liquid microextraction (DLLME) [327] among others. These techniques focus on reducing the time of sample preparation and the amounts of organic solvents. ...
Article
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Although the medicinal properties of Cannabis species have been known for centuries, the interest on its main active secondary metabolites as therapeutic alternatives for several pathologies has grown in recent years. This potential use has been a revolution worldwide concerning public health, production, use and sale of cannabis, and has led inclusively to legislation changes in some countries. The scientific advances and concerns of the scientific community have allowed a better understanding of cannabis derivatives as pharmacological options in several conditions, such as appetite stimulation, pain treatment, skin pathologies, anticonvulsant therapy, neurodegenerative diseases, and infectious diseases. However, there is some controversy regarding the legal and ethical implications of their use and routes of administration, also concerning the adverse health consequences and deaths attributed to marijuana consumption, and these represent some of the complexities associated with the use of these compounds as therapeutic drugs. This review comprehends the main secondary metabolites of Cannabis, approaching their therapeutic potential and applications, as well as their potential risks, in order to differentiate the consumption as recreational drugs. There will be also a focus on the analytical methodologies for their analysis, in order to aid health professionals and toxicologists in cases where these compounds are present.
... A number of works in the literature report the determination of THC and CBD concentration in hemp seed oil (Bosy and Cole, 2000;Leizer et al., 2000;Lachenmeier et al., 2004), but, to the best of our knowledge, there is no study regarding the evaluation of the comprehensive cannabinoid profile in this cannabis product. ...
Article
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Hemp seed oil is well known for its nutraceutical, cosmetic and pharmaceutical properties due to a perfectly balanced content of omega 3 and omega 6 polyunsaturated fatty acids. Its importance for human health is reflected by the success on the market of organic goods in recent years. However, it is of utmost importance to consider that its healthy properties are strictly related to its chemical composition, which varies depending not only on the manufacturing method, but also on the hemp variety employed. In the present work, we analyzed the chemical profile of ten commercially available organic hemp seed oils. Their cannabinoid profile was evaluated by a liquid chromatography method coupled to high-resolution mass spectrometry. Besides tetrahydrocannabinol and cannabidiol, other 30 cannabinoids were identified for the first time in hemp seed oil. The results obtained were processed according to an untargeted metabolomics approach. The multivariate statistical analysis showed highly significant differences in the chemical composition and, in particular, in the cannabinoid content of the hemp oils under investigation.
... The analysis can be performed by several methods, but the most widely employed are gas chromatography coupled to flame ionization detector (GC-FID) or mass spectrometry detector (GC-MS) and high performance liquid chromatography coupled to UV (HPLC-UV) or mass spectrometry detector (HPLC-MS) [17]. In particular, very few works have been published on the analysis of cannabinoids in hemp food products, most of which carried out by GC-MS [18][19][20][21]. The present work focuses on the development and validation of a rapid and sensitive HPLC-UV for the identification and quantification of the main cannabinoids, namely CBDA, THCA, CBD, THC, CBG, CBN and CBDV, in commercially available hemp seed oils. ...
Article
Hemp seed oil from Cannabis sativa L. is a very rich natural source of important nutrients, not only polyunsaturated fatty acids and proteins, but also terpenes and cannabinoids, which contribute to the overall beneficial effects of the oil. Hence, it is important to have an analytical method for the determination of these components in commercial samples. At the same time, it is also important to assess the safety of the product in terms of amount of any psychoactive cannabinoid present therein. This work presents the development and validation of a highly sensitive, selective and rapid HPLC-UV method for the qualitative and quantitative determination of the main cannabinoids, namely cannabidiolic acid (CBDA), tetrahydrocannabinolic acid (THCA), cannabidiol (CBD), tetrahydrocannabinol (THC), cannabinol (CBN), cannabigerol (CBG) and cannabidivarin (CBDV), present in 13 commercial hemp seed oils. Moreover, since decomposition of cannabinoid acids generally occurs with light, air and heat, decarboxylation studies of the most abundant acid (CBDA) were carried out in both open and closed reactor and the kinetics parameters were evaluated at different temperatures in order to evaluate the stability of hemp seed oil in different storage conditions.
... They concluded that the best preparation method for medical cannabis oil was with olive oil, since it is cheap, nontoxic, and nonflammable. The analysis of silylated cannabinoids (Δ9-THC-TMS, CBD-2TMS, CBN-TMS, and CBC-TMS) was performed on extracts from different matrices by Madea and coworkers [62,63]. They developed a fully automated method for the analysis of cannabinoids in hair samples and hemp food, using HS solid-phase dynamic extraction coupled with GC-MS (Fig. 4). ...
Article
Cannabis has garnered a great deal of new attention in the past couple of years in the United States due to the increasing instances of its legalization for recreational use and indications for medicinal benefit. Despite a growing number of laboratories focused on cannabis analysis, the separation science literature pertaining to the determination of cannabis natural products is still in its infancy despite the plant having been utilized by humans for nearly 30 000 years and it being now the most widely used drug world-wide. This is largely attributable to the restrictions associated with cannabis as it is characterized as a Schedule 1 drug in the United States. Presented here are reviewed analytical methods for the determination of cannabinoids (primarily) and terpenes (secondarily), the primary natural products of interest in cannabis plants. Focus is placed foremost on analyses from plant extracts and the various instrumentation and techniques that are used, but some coverage is also given to analysis of cannabinoid metabolites found in biological fluids. The goal of this work is to provide a collection of relevant separation science information, upon which the field of cannabis analysis can continue to grow.
... However, it should be noted, that the highest yields were obtained at maximal τ value; therefore, optimisation in this case means determination of optimal parameters in the selected range of variables. For comparison, previously reported CBD content in commercially available hemp seed oils was 4-236 mg/kg and hemp-leaf containing herbal teas 26-60 mg/kg (Lachenmeier, Kroener, Musshoff, & Madea, 2004;Petrović, Debeljak, Kezić, & Džidara, 2015). ...
Article
C. sativa threshing residues were biorefined by consecutive supercritical carbon dioxide (SFE-CO2) pressurised liquid (PLE) and enzyme-assisted extractions (EAE). SFE-CO2 at optimised parameters yielded 8.3g/100g of lipophilic fraction containing 0.2 and 2.2g of cannabidiol and cannabidiolic acid per 100g of threshing residues, respectively. The recovery of cannabinoids from plant material was >93%. PLE gave 4.3 and 18.9g/100g of flavonoid-containing polar extracts, while EAE added 20.2% (w/w) of water-soluble constituents and increased the release of mono- and disaccharides by up to 94%. Antioxidant capacity of non-polar and polar fractions was in the range of 1.3-23.5mg gallic acid equivalents/g DW and 0.6-205.2mg Trolox equivalents/g DW, with the highest activities of PLE-EtOH/H2O extract. The combined SFE-CO2, PLE and EAE reduced antioxidant capacity of starting plant material by 90-99%, showing that suggested multistep fractionation procedure is efficient in the recovery of a major part of the antioxidatively active constituents from hemp threshing residues.
... In addition, these fibers are resistant to bacteria, mold, and ultraviolet rays, and they can be used in healthcare applications (Mohanty et al. 2002;Keller 2003;Khan et al. 2014). With the low content of tetrahydrocannabinol (THC < 0.3%) that has been achieved by breeding of hemp varieties (Lachenmeier et al. 2004), hemp has gradually come to the forefront of textile applications. The hemp fiber can be blended and spun with cotton, silk, wool, and other chemical fibers, and it can also be spun into mono-fibers. ...
Article
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This bio-chemical study focuses on obtaining high-quality hemp fiber. The effects of the structures and properties of hemp fibers in different treatment periods were studied. Moreover, the changes of the surface morphology, chemical composition, and breaking tenacity of hemp fibers were researched by scanning-electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), fluorescence microscopy, and fiber tensile testing. The results showed that by virtue of the enzyme scouring process, alkali refining process, and bleaching process, the pectin, lignin, and hemicellulose and other impurities were removed. Through the single factor experiment, the optimal process conditions for the bio-chemical combination of the degumming process were obtained. These conditions included 10 g of dried hemp fibers, 15% (v/v) pectinase solution, a temperature of 50 °C, a duration of 120 min, pH 8.0 (phosphate buffer), a liquor ratio (w/v) of 1:10, and 0.0625 mol/L NaOH. In these conditions, the residual gum content and breaking tenacity were 4.8% and 49.8 cN/tex, respectively, indicating that the treated hemp fibers met the requirements of the spinning process.
... For this purpose, we chose solid phase microextraction [12] coupled to headspace sampling, which is based on the adsorption of the volatile analytes by the coating of a suitable fiber and their direct injection into a GC/MS system. This method shows several advantages respect to liquid-liquid extraction (LLE), because it is more selective, less time consuming, it does not require the use of solvents and it is particularly suitable for volatile analytes [13]. Different parameters of the analytical method (fiber, coating thickness, sampling and exposition temperatures, sample preparation) were evaluated to optimize it for the determination of the volatile components of hashish, in particular, the parameters were studied taking into account azulene and caryophyllene, two significant representatives of the monoterpene and sesquiterpene classes present in hashish (Figure 1). ...
Article
Solid phase microextraction coupled to headspace sampling and GC/MS technique was applied to the characterization of the volatile components of several Cannabis preparations (hashish). Different parameters of the analytical method (fiber, coating thickness, sampling and exposition temperatures, sample preparation) were evaluated to optimize the characterization of the volatile components. a-Pinene, ß-myrcene, limonene, 4-carene, trans-3(10) caren-2-ol, 4,7,7-trimethylbicyclo [4.1.0] heptan-3-ol, caryophyllene, ß-humulene, azulene, gurjunene, ledene and caryophyllene oxide were identified among the volatile components of all hashish preparations. Moreover, a suitable internal standard (nonane) was chosen, the reproducibility and linearity of the method were evaluated in order to carry out the quantitative determination of caryphyllene, the most abundant volatile terpene. Its quantity ranged from 800 to 3000 µg/g.
... Hemp (Cannabis sativa) is a plant able to synthetize more than 60 cannabinoids being the main active component the D9tetrahidrocannabinol (THC), followed by the cannabidiol and the cannabinol (Lachenmeier, Kroener, Musshoff, & Madea, 2004). The hemp varieties allowed for cultivation in Europe have less than 0.2% THC, which is mostly present as D9-tetrahydrocannabinol acid (THC-A) a non-psychotropic constituent that account for 90% of total cannabinols in fiber-type cannabis plant (EFSA, 2011;Grotenhermen, 2003;Huestis, 2007;Takeda et al., 2012). ...
Article
A method for determining cannabinoids, Δ9-tetrahidrocannabinol (THC), 11-nor-9-carboxy-Δ⁹-THC (THC-COOH) and 11-hidroxy-Δ⁹-THC (THC-OH) in milk, liver and hemp seeds based on liquid chromatography tandem mass spectrometry has been optimized and validated. Analytes were extracted with methanol and the extracts cleaned-up by solid-phase extraction using Oasis HLB (60 mg). The developed method was validated according to the Commission Decision 2002/657/EC. The decision limit (CCα) and detection capability (CCβ) ranged from 3.10 – 10.5 ng g⁻¹ and 3.52 – 11.5 ng g⁻¹, the recoveries were 76 – 118% and matrix effect ranged from -17.8% to 19.9% in the three matrices studied. The method was applied to food samples obtaining positive results for THC in hemp seeds (average 0.82 μg g⁻¹) and three brands of junior formula milk at concentrations from 4.76 to 56.11 ng g⁻¹. The developed method was suitable achieving identification and quantification of cannabinoids in food matrices.
... Niobic acid made from the niobic oxide showed very good water-tolerant characteristics and good performance in canalization of many reactions in water (Guo et al., 2003;Lachenmeier, Kroener, Musshoff, & Madea, 2004). In this paper, various mineral acids coating with niobic acid were prepared with different preparation conditions such as impregnation time, calcination temperature and calcination time and different acids (e.g. ...
... The performance of a validated Category A requires just one other technique (from Category A, B or C) to complement the forensic analysis. Chromatographic techniques (B) are usually employed during analyses of illicit drugs (10)(11)(12), and a combination with mass spectrometry (A) constitutes a valuable tool to analyze traces and their metabolites (13,14). If a Category A test is not available at the time of analysis, the analyst must use three different validated techniques and two B tests accompanied by a third complementary test that can belong to Category B or C (7,15). ...
Article
The development of methodologies using inexpensive, fast, and reliable instrumention is desirable in illicit drug analysis. The purpose of this study was based on cyclic voltammetry technique to differentiate the electrochemical behavior of ∆9-THC, the psychoactive substance in marijuana, and five different extract plants to yield false positive results after analysis protocol for cannabinoids using thin-layer chromatography and Fast Blue B salt. After applying a deposition potential of −0.5 V in a glassy carbon working electrode, the results indicated an anodic peak current at 0.0 V versus Ag/AgCl after addition of ∆9-THC solution in the electrochemical cell, and limits of detection and quantification were 1.0 ng mL−1 and 3.5 ng mL−1, respectively. Other interfering plants showed distinct amperometric responses. This methodology was useful to detect ∆9-THC even in the presence of the Fast Blue B salt, which avoided false positive results for all the studied extract plants.
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Kenevir ve endüstri
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The fibrous hemp Cannabis sativa L. called also: industrial hemp and cannabis, is a plant widespread all over the world, grown mainly for its fiber, used for various purposes. Hemp fruits (also known as seeds or nuts) are mainly used to produce of oil for food, cosmetic and as bird feed. The nuts contain about 30–35% of oil with 80 to 90% of essential fatty acids (EFA), of which 55–56% is linoleic acid (LA) from the omega-6 group and 22–25% α-linolenic acid (ALA) from the omega-3 group, as well as amino acids, participating in the formation of albumin and globulin (edestin), vitamins, and minerals. The article describes in detail: the nutritional value of hemp nuts, the possibilities for the use of hemp nuts, discussion about the cannabinoid content of fibrous hemp nuts, and legislative issues concerning the cultivation of industrial hemp. The paper reviews the literature on the most important nutritional properties of the seeds and products made from the processing of hemp seeds, making it a very versatile raw material used among others in the food industry.
Article
Although still illegal in many countries, food products containing cannabis or marijuana extracts have become very popular in recent years. In the present study, an LC-MS method was developed for the quantitative analysis of seven cannabinoids in various solid and liquid cannabis-based goods. The proposed analytical approach demonstrated satisfactory performance characteristics in terms of linearity (R²≥0.995), accuracy (recovery: 70.0-110%), precision (intraday RSD: 0.950-6.03%, interday RSD: 1.02-6.94%), sensitivity (LOD≤2.19 ng/mL, LOQ≤6.59 ng/mL) and carry-over effect (average carryover signals ≤3.90%). Solid-phase extraction (SPE), and ultrasound-assisted extraction (UAE) were utilized for the extraction of the analytes from liquid cannabis edibles (beer and energy drink), while Soxhlet and ultrasound-assisted extraction (UAE) were used for solid products (chocolates, hemp seeds, hemp tea). Infusion and decoction processes were followed for cannabis hemp tea and roasted coffee, respectively. UAE provided higher extraction efficiencies for cannabis-based edibles in solid form, while infused-cannabis beverages were extracted more efficiently using the SPE procedure. Cannabidiol (CBD) and cannabigerol (CBG) were the most detectable cannabinoids in all examined samples. Significantly high levels of cannabinoids were detected in cannabis tea extract prepared by the UAE procedure (total cannabinoids: 5440 μg/g). According to the suppliers, all examined samples were supposed to be free of Δ⁹-tetrahydrocannabinol (Δ⁹-THC). However, five products were found to contain considerable amounts of this compound (0.600-180 μg/g). Only in the case of cannabis beer, cannabis roasted coffee, and cannabis energy drink, Δ⁹-THC was not detected.
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A simple quantitative reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for assay determination of cannabidiol and tetrahydrocannabinol in hemp oil infused products. The RP-HPLC method was developed and optimized for the mobile phase composition, flow rate, column selection and detector wavelength. An isocratic elution of samples were performed on SOLAS 100 Å C18 150 mm × 4.6 mm, 5 μm column with a mobile phase containing 75/25 acetonitrile/water v/v, with a flow rate of 1.5 mL/min by using an ultraviolet–visible (UV/Vis) detector operating at 214 nm. The RP-HPLC method was validated to meet regulatory requirements which covers specificity, accuracy, range, linearity, precision, system suitability and robustness. The validated assay test method was applied successfully to quantify cannabidiol and tetrahydrocannabinol in commercial hemp oil infused products such as tablets, soft gel capsules, plant extract oils, oral drops, tincture, and beverage enhancers. All the test results were found acceptable as per ICH guidelines, and this confirmed the feasibility of this method for its intended use in regular quality control and assay of cannabidiol and tetrahydrocannabinol in hemp oil infused products.
Article
Hemp is one of the most complete plants for industrial and consumer purposes. In pharmaceutical industry, cannabidiol (CBD) extracts are gaining increasing attention due to their therapeutic properties. There is a lack of information about their inorganic constituents, as well as assessment of these elements that are essential and/or potentially toxic towards humans. The inorganic elements quantified in the hemp samples by ICP OES were: Mg (5–8000); Ca (10–1780); P (39–17500) and K (6500–14000) mg kg⁻¹. The microelements obtained were: Mo (0.03–1.1); Ba (0.09–2.4); Sr (0.21–7.8); Cr (0.30–0.62); Ni (0.81–1.0); Na (1.4–11.1); Cu (19.5–24.0); Mn (0.66–152); Zn (51–96) and Fe (111–168) mg kg⁻¹. The elements levels are related to the manufacturing processes of the hemp products, being below the tolerable upper daily intake levels. For the seeds samples, the inorganic constituents levels were: Ca (313–1164); Mg (4498–6734); K (7500–12104) and P (9623–13636) mg kg⁻¹. The microelements levels were: Mo (0.46–1.3); Cr (0.48–1.9); Ba (0.48–11.0); Ni (0.66–3.7); Sr (4.6–22.0); Cu (10.2–14.2); Na (11.1–87.0); Mn (38−83); Zn (62–82) and Fe (92–112) mg kg⁻¹. Hemp seeds also represent an excellent source of trace elements essential to health. The CBD extracts, showed low levels of inorganic constituents, that there was no risk to human health. The nutritional order established for the Cannabis-based products as it follows: hemp protein > shelled seeds > peeled seeds > hemp butter > hemp oil and CBD extracts.
Article
Cannabis sativa L. is an intriguing plant that has been exploited since ancient times for recreational, medical, textile and food purposes. The plant's most promising bioactive constituents discovered so far belong to the terpenoid and cannabinoid classes. These specialised metabolites are highly concentrated in the plant aerial parts and their chemical characterisation is crucial to guarantee the safe and efficient use of the plant material irrespective of which use it is. This study investigates for the first time the use of vacuum assisted HS-SPME as a sample preparation process in an analytical protocol based on HS-SPME combined to fast GC-MS analysis that aims at comprehensively characterising both the terpenoid and cannabinoid profiles of Cannabis inflorescences in a single step. The results proved that vacuum in the HS should be preferred over atmospheric pressure conditions as it ensures the fast recovery of cannabinoid markers at relatively lower sampling temperatures (i.e., 90°C) that do not discriminate the most volatile fraction nor cause the formation of artefacts when the sampling time is minimised.
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A simple quantitative reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for assay determination of cannabidiol and tetrahydrocannabinol in hemp oil infused products. The RP-HPLC method was developed and optimized for the mobile phase composition, flow rate, column selection and detector wavelength. An isocratic elution of samples performed on SOLAS™ 100 Å C18 150 mm x 4.6 mm, 5 µm column with a mobile phase containing 75/25 acetonitrile/water v/v, delivered at a flow rate 1.5 mL/minutes to an Ultraviolet – Visible (UV/Vis) detector using 214 nm. The RP-HPLC method was validated to meet regulatory requirements and covers specificity, accuracy, range, linearity, precision, system suitability and robustness. The validated assay test method was applied successfully to quantify cannabidiol and tetrahydrocannabinol in commercial hemp oil infused products such as tablets, soft gel capsules, plant extract oils, oral drops, tincture, and beverage enhancers. All test results were found acceptable as per ICH guidelines, and this confirmed that the method is fit for its intended use in regular quality control and assay of cannabidiol and tetrahydrocannabinol in hemp oil infused products.
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An interesting and valuable discussion has arisen from our recent article (Lachenmeier et al., 2020) and we are pleased to have the opportunity to expand on the various points we made. Equally important, we wish to correct several important misunderstandings that were made by Kruse and Beitzke (2020) on behalf of the European Industrial Hemp Association (EIHA) that possibly contributed to their concerns about the validity of our data, toxicological assessment and conclusions regarding regulatory status of cannabidiol (CBD) products. First and foremost, our study did only assess the risk of psychotropic Δ ⁹ -tetrahydrocannabinol (THC) without inclusion of non-psychotropic Δ ⁹ -tetrahydrocannabinolic acid (THCA). Secondly, as this article will discuss in more detail, there is ample evidence for adverse effects of CBD products, not only in paediatric patients, but also in adult users of over-the-counter CBD products (including inadvertent “high” effects). Thirdly, the exposure and risk assessment was conducted using up-to-date guidelines according to the European Food Safety Authority (EFSA) and the German Federal Institute for Risk Assessment (BfR). And finally, the current legal situation in the European Union, without approval of any hemp extract-containing product according to the Novel Food regulation, actually allows blanket statements that all such products are illegal on the market, and this indeed would imply a general ban on the use and marketing of such products as food or food ingredients until such an approval has been granted. We hope that this reassures the F1000Research readership regarding the validity of our results and conclusions. We are pleased, though, that the EIHA has acknowledged the fact that there are non-compliant CBD products available, but according to our data these are a substantial fraction of the market.
Article
In the past, a variety of herbs were used in brewing, however only hops (Humulus lupulus) are now widely used as they contribute to the bitterness, flavour and microbiological stability of beer. After the abolition of prohibition of the cultivation of the Cannabis sativa L. species (the closest relative of H. lupulus), there are now beers infused with cannabis extracts made from all parts of the plant. The variety ‘hemp’ is used as it contains a minor concentration of the psychoactive Δ9‐tetrahydrocannabinol compared with marijuana. In this review, H. lupulus and C. sativa are compared and the opportunities and constraints for producing cannabis beers are discussed. © 2021 The Institute of Brewing & Distilling
Article
Herbal teas of fiber-type hemp varieties (Cannabis sativa L.) rich in cannabidiolic acid (CBDA) and cannabidiol (CBD) and poor in Δ9-tetrahydrocannabinol acid (THCA) and Δ9-tetrahydrocannabinol (THC) are very popular today. The conditions for preparing herbal infusions are not well standardized and analysis of the lipophilic cannabinoids in infusions is difficult. Therefore, we analyzed the hemp leaf residues after tea preparation by using a response surface modelling approach to estimate the effects of variations in temperature, water volume and extraction time on the residual content of five cannabinoids (CBDA, CBD, THCA, THC, cannabinol (CBN)) in the hemp leaves after extraction. The quantity of remaining cannabinoids was mainly influenced by temperature in the first order. Volume and extraction time were only exerting minor influences under usual tea preparation processes. At elevated water temperatures CBD and THC values were even higher than in the original drug material presumably due to decarboxylation of CBDA and THCA. Rising temperatures increased extraction of CBDA and CBD, as opposed to THCA and THC. The degradation of THC to CBN was not significant at the conditions of infusion preparation. Analyzing herbal residues after tea brewing is just an approximation to the true values of valuable or unfavorable compounds in tea, overestimating the true values. However, that approach offers a good control for further improvements of herbal tea analysis and gives reliable indications for risk assessment.
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Mead, one of the oldest existing drinks, is a fermented product based on honey, water, and the possible addition of spices and selected yeasts. In this work, various parts (inflorescences, leaves, and steams) of Cannabis sativa L. at different concentrations and Saccharomyces cerevisiae biotype M3/5 were added during mead fermentation. The physicochemical parameters (pH, alcoholic content, sugar content, titratable acidity, and organic acids) of the mead were assessed at the beginning and end of fermentation. Moreover, polyphenols, cannabidiol and volatile organic compounds were identified at the end of fermentation and compared with the control sample prepared without hemp and with only indigenous yeasts. The mead fermented with hemp showed the highest quantity of polyphenols (227 to 256 mg GAE/L) and a level of cannabidiol ranging from 0.26 to 0.49 mg/kg. The volatile organic compounds found were mainly alcohols, esters and terpenes, which were present at higher concentrations in the mead prepared with C. sativa L. than in the control mead and conferred freshness and “hemp aroma” characteristics. Practical Application Inflorescences, leaves, and steams of Cannabis sativa L. were added at different concentrations during mead fermentation. This type of mead showed high quantity of polyphenols (227 to 256 mg GAE/L) and a level of cannabidiol ranging from 0.26 to 0.49 mg/kg which have anxiolytic and neuro‐protective properties. Moreover the volatile organic compounds found (mainly alcohols, esters, and terpenes) conferred freshness and “hemp aroma” characteristics.
Conference Paper
Many false positive and false negative results have been detected in immunoassay analyses of drugs of abuse in urine samples. A method of direct injection of diluted urine into LC/MS/MS was developed and validated for detection and quantitation of Amphetamine, Methamphetamine, MDMA, MDA, Benzoylecgonine, Ecgonine, Norpseudoephedrine, Ephedrine, Tapentadol, Tramadol, O-desmethyltramadol, Tapentadol, Pregabline, Gabapentine and Methadone to avoid the false positive and false negative results in urine samples. Linearity of Amphetamine, Methamphetamine MDMA, MDA, Benzoylecgonine, Ecgonine, Norpseudoephedrine and Ephedrine was (60-2400ng/mL), for Tapentadol, Tramadol, O-desmethyltramadol, and Methadone was (50-1600 ng/mL), and for Pregabline and Gabapentine was (100-4000ng/mL) and r2 ˃ 0.992 for all analysts. A 440 urine samples have been analyzed using both immunoassay technique and LC/MS/MS by direct injection method giving a good comparison to illustrate how this method was specific, accurate, precise, and applicable for forensic urine samples
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The use of Cannabis sativa oils has been growing in the last years as an emerging therapy for non-responsive clinical conditions. Some benefts have been noticed, mainly in neurological diseases such as multiple sclerosis, epilepsy, psychosis, depression, and anxiety [1, 2]. Its applications also include the reduction of adverse efects in cancer therapy and the treatment of chronic pain [3]. Moreover, the pathways related to the cannabis benefts are still being discovered [4]. The therapeutic properties can be correlated to the presence of cannabidiol (CBD), although the pharmacological activity can be infuenced by other phytocannabinoids found in the plant. However, high concentrations of Δ9 -tetrahydrocannabinol (THC) and cannabinol (CBN) can be associated with several side efects
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In this study, a simple and rapid method for screening synthetic cannabinoids in herb mixtures and human blood by gas chromatography-mass spectrometry (GC/MS) with headspace solid-phase microextraction (HS-SPME) was developed. Herb mixtures were taken into an SPME vial without any pretreatment. Blood samples were pre-incubated with cholesterol oxidase and subjected to deproteinization with acetonitrile, and the liquid phase was then transferred into an SPME vial and purged to dryness with nitrogen. These samples were analyzed by HS-SPME-GC/MS. The fiber material used for the SPME was polyacrylate, and a DB-5 ms capillary column was used for the GC. The analysis of the combustion residues of herbal mixtures considered to have been used by a drug abuser revealed that a certain synthetic cannabinoid, so called AB-CHMINACA was included in the sample. Although cholesterol in blood interfered with the measurement of one synthetic cannabinoid (JWH-015), it was possible to eliminate the cholesterol beforehand by oxidation with cholesterol oxidase. This study demonstrates an applicability of a convenient, rapid and highly useful screening method of illegal herbal mixtures and blood specimens.
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CBD treats for dogs CBD (cannabidiol) is a subject of topical interest to many dog owners, fuelled by an increasing supply of CBD-infused treats. These products are claimed to mitigate inflammatory skin and joint problems, anxiety and epileptic seizures (Note 1). CBD is derived from industrial hemp crops low in the psychoactive and addictive THC (tetrahydrocannabinol), which is abundant in other hemp plants, known as marijuana. A couple of complete dog foods contain hemp seed or its oil fraction. CBD dog snacks generally include CBD oils, oily extracts from hemp inflorescences mixed with the seeds' oil or another oil carrier. The CBD content of CBD oils varies markedly, while THC may be absent or substantial. Hemp whole seeds and seed oil are mostly low in CBD and THC, but a single oil may be CBD-rich. Manufacturers must allow for the variable composition of CBD oils in order to supply treats with CBD (and THC) contents that permanently meet their pre-set targets. There are no available data on CBD and anxiety or inflammatory skin disease in dogs. Researching CBD's impact on joint disease and epilepsy is underway (1). As to epilepsy, preliminary affirmation has been posted (2). One well-performed study, published recently, gave conflicting results for the efficacy of CBD in dogs with joint disease. The dogs suffering from arthritis were administered CBD oil by mouth. A 20-kg dog would receive 80 mg pure CBD per day. CBD oils have complex compositions, not least due to the big family of cannabinoid compounds. The naming implies that CBD is considered the chief active principle. Commercial CBD treats carry feeding instructions and may declare the amount of CBD per treat. When taking the mean for six products, a 20-kg dog gets 10 mg CBD per day. This consumption level appears safe, but does not justify health claims, at least for the time being.
Article
Rationale Phytocannabinoids are natural compounds produced by Cannabis spp. Some of them show psychotropic effects on humans and are therefore are used as drugs of abuse. These compounds are present in food and beverages containing ingredients from hemp, and thus can reach consumers. The Italian Ministry of Health planned to evaluate the intake of cannabinoids from food containing hemp ingredients. Thus, we were asked to develop and validate a multi‐residue test method to determine nine phytocannabinoids in beverages and food. Methods Nine natural phytocannabinoids, hereafter called cannabinoids, were cleaned up from food by solid‐liquid extraction, while beverages were simply diluted prior to analysis. The cannabinoids were separated by reversed phase high performance liquid chromatography, and on‐line determination was carried out by tandem mass spectrometry using a 4000 QTRAP mass spectrometer with a TurboIonSpray source, in multiple reaction‐monitoring mode, using both positive and negative ionization. Results Each compound was determined down to 0.25 ng/mL in solvent. In‐house validation was carried out; the mean recoveries ranged from 83.4% to 101.2% in food, and from 84.5% to 104.5% in beverages. The limits of quantification were 20 μg/kg for food and 2 μg/L for beverages. Conclusions A reliable and rapid method for the identification and quantification of the psychotropic Δ⁹‐tetrahydrocannabinol, its non‐psychoactive precursor Δ⁹‐tetrahydrocannabinolic acid A, and seven other cannabinoids was developed and validated, to monitor the content of these substances in food and beverages produced using hemp seeds, flour and oil as ingredients.
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A routine method for determining cannabinoids in Cannabis sativa L. inflorescence, based on Fast gas chromatography coupled to mass spectrometry (Fast GC/MS), was developed and validated. To avoid the decarboxylation of carboxyl group of cannabinoids, different derivatization approaches, i.e. silylation and esterification (diazomethane-mediated), reagents and solvents (pyridine or ethyl acetate), were tested. The methylation significantly increased the signal-to-noise ratio of all carboxylic cannabinoids, except for cannabigerolic acid (CBGA). Since diazomethane is not commercially available, is considered a hazardous reactive and requires 1-day synthesis by specialized chemical staff, silylation was used along the whole validation of a routine method. The method gave a fast (total analysis time < 7.0 min) and satisfactory resolution (R > 1.1), with a good repeatability (intraday < 8.38%; interday < 11.10%) and sensitivity (LOD < 11.20 ng/mL). The Fast GC/MS method suitability for detection of cannabinoids in hemp inflorescences, was tested; a good repeatability (intraday < 9.80%; interday < 8.63%), sensitivity (LOD < 58.89 ng/mg) and robustness (<9.52%) was also obtained. In the analyzed samples, the main cannabinoid was cannabidiolic acid (CBDA, 5.19 ± 0.58 g/100 g), followed by cannabidiol (CBD, 1.56 ± 0.03 g/100 g) and CBGA (0.83 g/100 g). Δ9-tetrahydrocannabivarine (THCV) was present at trace level. Therefore, the developed routine Fast GC/MS method could be a valid alternative for a fast, robust and high sensitive determination of main cannabinoids present in hemp inflorescences.
Article
Headspace solid phase microextraction (HS-SPME), which is a solvent-free extraction technique, was configured with gas chromatography/mass spectrometry (GC/MS) to detect phytocannabinoids from buccal swabs. The HS-SPME extraction procedure, i.e. extraction time, extraction temperature, thermal desorption parameters as well as headspace derivitization, were evaluated to extract major phytocannabinoids from the headsapce of air-dried buccal swab samples. Sub micrograms of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) spikded onto buccal swabs could be extraced and detected by the HS-SPME-GC/MS appraoch. The analytical system can be readily automated without the use of solvent extraction. No interference peaks for phytocannabinoids were found in the total ion chromatograms obtained from the tested buccal swabs using cotton as the substrate. Interference background can also be minimized by using selected ion monitoring. This analytical approach potentially could be adopted to detect marijuana smokers by the identification of residual phytocannabinoids from oral cavities for forensic applications.
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The fatty acid and natural product content of hemp seed oil was analyzed by GC-MS and LC-MS. The presence of linoleic acid (LA) and -linolenic acid (LNA) were confirmed in their previously reported ratio of 3:1 LA:LNA. The presence of -caryophyllene (740 mg/L), myrcene (160 mg/L), -sitosterol (100-148 g/L) and trace amounts of methyl salicylate was observed in the oil which had not been previously reported. Trace amounts of cannabidiol (CBD) were also detected. Bioassays were performed with the oil to determine its effectiveness as an antimicrobial agent. Some bioactivity was observed during the primary screening. (Article copies available for a fee from The Haworth Document Delivery Service: 1-800-342-9678. E-mail address: Website: )
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A commercially available health food product of cold-pressed hemp seed oil was ingested by one volunteer twice a day for 4½ days (135 mL total). Urine specimens collected from the volunteer were subjected to standard workplace urine drug testing procedures, and the following concentrations of 11-nor-Δ9-tetrahydrocannabinol carboxylic acid (9-THCA) were detected: 41 ng/mL 9-THCA at 45 h, 49 ng/mL at 69 h, and 55 ng/mL at 93 h. Ingestion was discontinued after 93 h, and the following concentrations were detected: 68 ng/mL at 108 h, 57 ng/mL at 117 h, 31 ng/ml at 126 h, and 20 ng/ml, at 142 h. The first specimen that tested negative (50 ng/mL initial immunoassay test, 15 ng/mL confirmatory gas chromatographic-mass spectrometric test) was at 146 h, which was 53 h after the last hemp seed oil ingestion. Four subsequent specimens taken to 177 h were also negative. This study indicates that a workplace urine drug test positive for cannabinoids may arise from the consumption of commercially available cold-pressed hemp seed oil.
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Gas chromatographic-mass spectrometric (GC-MS) quantitation of 25 cannabis sed oils determined delta 9-tetrahydrocannabinol (THC) concentrations from 3 to 1500 micrograms/g oil. In a pilot study, the morning urine of six volunteers who had ingested 11 or 22 g of the oil, which contained the highest THC content (1500 micrograms/g), was collected for six days. The urine samples were screened by immunoassay, and the content of 11-nor-9-carboxy-delta 9-THC (THCCOOH) was determined by GC-MS. Urine samples were found cannabis positive for up to six days with THCCOOH-equivalent concentrations up to 243 ng/mL. by the Abuscreen OnLine immunoassay and THCCOOH contents from 5 to 431 ng/mL by the GC-MS method. All subjects reported THC-specific psychotropic effects.
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A hemp oil product (Hemp Liquid Gold) was purchased from a specialty food store. Fifteen milliliters was consumed by seven adult volunteers. Urine samples were taken from the subjects before ingestion and at 8, 24, and 48 h after the dose was taken. All specimens were screened by enzyme immunoassay with SYVA EMIT II THC 20, THC 50, and THC 100 kits. The tetrahydrocannabinol carboxylic acid (THCA) concentration was determined on all samples by gas chromatography-mass spectrometry (GC-MS) (5). A total of 18 postingestion samples were submitted. Fourteen of the samples screened above the 20-ng cutoff, seven were above the 50-ng cutoff, and two screened greater than the 100-ng cutoff. All of the postingestion samples showed the presence of THCA by GC-MS.
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Commercially available snack bars and other foodstuffs prepared from pressed hemp seeds were ingested by volunteers. Urine specimens were collected for 24 h after ingestion of the foodstuffs containing hemp seeds and tested for marijuana using an EMIT immunoassay and gas chromatography-mass spectrometry (GC-MS). Specimens from individuals who ate one hemp seed bar demonstrated little marijuana immunoreactivity, and only one specimen screened positive at a 20-ng/mL cutoff. Specimens from individuals who ate two hemp seed bars showed increased immunoreactivity, and five specimens screened positive at a 20-ng/mL cutoff. A single specimen yielded a quantitative GC-MS value (0.6 ng/mL), but it failed to meet reporting criteria. Several specimens from individuals who ate three cookies made from hemp seed flour and butter screened positive at both 50- and 20-ng/mL cutoffs. Two specimens produced quantitative GC-MS values (0.7 and 3.1 ng/mL), but they failed to meet reporting criteria. Several specimens also tested positive with an FDA-approved on-site marijuana-screening device. Hemp seeds similar to those used in the foodstuffs did not demonstrate the presence of marijuana when tested by GC-MS. In this study, ingestion of hemp seed food products resulted in urine specimens that screened positive for marijuana. No specimens gave a GC-MS quantitative value above the limit of detection for marijuana.
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This paper describes the application of solid-phase microextraction (SPME) to cannabis testing in hair. Fifty milligrams of hair was washed with petroleum ether, hydrolyzed with NaOH, neutralized, deuterated internal standard was added and directly submitted to SPME. The SPME was analyzed by GC-MS. The limit of detection was 0.1 ng/mg for cannabinol (CBN) and delta9-tetrahydrocannabinol (THC) and 0.2 ng/mg for cannabidiol (CBD). THC was detected in a range spanning from 0.1 to 0.7 ng/mg. CBD concentrations ranged from 0.7 to 14.1 ng/mg, and CBN concentrations ranged from 0.4 to 0.7 ng/mg. The effectiveness of different decontamination procedures was also studied on passively contaminated hair. The proposed method is also suitable for the analysis of methadone in hair; cocaine and cocaethylene can be detected in hair with SPME extraction after enzymatic hydrolysis.
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Today, cannabis plants are used in shampoo preparations, in foodstuffs (e.g., oils, noodles, crackers, etc.), and in beverages (e.g., tea). These products often contain < 1% delta9-tetrahydrocannabinol (THC) in order to eliminate psychoactive effects, but some of them can include 1 to 3% of THC. Gas chromatography-mass spectrometry (GC-MS) analysis of Cannabio shampoo revealed the presence of THC (412 ng/mL) and two constituents of cannabis plants, cannabidiol (CBD, 4079 ng/mL) and cannabinol (CBN, 380 ng/mL). In order to verify if normal hygiene practices with Cannabio shampoo can result in positive tests for cannabinoids in hair, three subjects washed their hair with this shampoo once daily for two weeks. After this period, hair specimens were collected. In the three hair specimens, THC, CBD, and CBN were never detected within their limits of detection, 0.05, 0.02, and 0.01 ng/mg, respectively. We concluded that the use of Cannabio shampoo during normal hygiene practices cannot be considered as a source of potential contamination of hair. In a second experiment, drug-free hair specimens (200 mg) were incubated in 10 mL water/Cannabio shampoo (20:1, v/v) for 30 min, 2 h, and 5 h. After incubation, hair strands were washed with water and separated into two portions. One portion was extracted directly; the second was decontaminated with methylene chloride and then extracted. After an incubation period of 30 min, the analysis of hair by GC-MS did not reveal the presence of THC, CBD, and CBN in hair, regardless of whether the hair was decontaminated. After an incubation period of 2 h, specimens tested positive for CBD (0.11 ng/mg without decontamination and 0.10 ng/mg with decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). After an incubation period of 5 h, specimens tested positive for CBD (0.25 ng/mg without decontamination and 0.14 ng/mg after decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). In all cases, THC was never detected. Extensive but unrealistic use of Cannabio shampoo can cause drug-free hair to test positive for CBD and CBN but not for the primary psychoactive drug THC.
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A GC-MS method was performed to determine the total delta9-THC content in both drug- and fiber-type cannabis seeds. Drug-type seeds were found to contain much higher levels of delta9-THC (35.6-124 microg/g) than fiber (hemp) seeds (0-12 microg/g). The majority of delta9-THC was found to be located on the surface of the seeds. Approximately 90% of the total delta9-THC was removed by a simple, quick wash with chloroform. Washed drug-type seeds contained less than 10 microg/g. Separation of the seeds into the kernel and testa showed that the bulk of delta9-THC is located in the testa, mainly on the outside. The kernels of drug- and fiber-type cannabis seeds contained less than 2 and 0.5 microg delta9-THC/g seeds, respectively. Fluctuations in the delta9-THC content of different replicates of the same type of seeds could be the result of the degree of contamination on the outside of the seeds.
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Two types of oils containing hemp that are freely available on the market were shown to contain Tetrahydrocannabinol (THC) and Cannabidiol. In addition, one of the two also contained Cannabigerol and Cannabinol. A few hours after oral ingestion of the oils 11-nor-delta 9-THC-9-Carbonic acid, which is the main metabolite of delta 9-THC could be detected in the urine. After the ingestion of 40 to 90 ml of oil THC-carbonic acid could be found in the urine for up to 80 hours. After the ingestion of 40 ml hempseed oil a THC-serum level of up to 6 ng/ml could be detected in blood samples.
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A GC-MS method for the analysis of Δ9-tetrahydrocannabinol in food was developed. After extraction with n-hexane and saponification, Δ9- tetrahydrocannabinol was determined by gas chromatography-mass spectrometry in the unsaponifiable matter.
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Tetrahyrocannabinol and cannabinol were identified and determined in cannabis seeds sold as feed for birds by thin-layer chromatography, gas chromatography and gas chromatography-mass spectrometry.
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After previous examination of oils containing hemp a series of other foodstuff containing hemp was analysed. All products were freely available on the market. In all products Tetrahydrocannabinol (THC) could be detected. In some it was accompanied by Cannabinol and Cannabidiol. The THC content in urine were at such a level that, assuming normal consumption, they could not be detected in urine tests in all but one case. Particularly the examined hemp lequeur showed no effect whatsoever in terms of psychotropic influence. It did however have an intoxication influence due to the alcohol content.
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Nine strains of Cannabis sativa L. (marijuana) were grown for research by the University of Mississippi. The seeds for these strains were obtained from Iowa, Minnesota, Mexico, Turkey, Italy, France, and Sweden. The cannabinoid content was determined using GLC, and the material was divided into two chemical phenotypes according to cannabinoid content. These phenotype categories are used to differentiate between drug-type and fiber-type Cannabis sativa. In addition, the ( - )-δ9−trans-tetrahydrocannabinol content was determined for both male and female plants, various plant parts, and a Turkish variety during various stages in its growth.
Article
Food analysis is important for the evaluation of the nutritional value and quality of fresh and processed products, and for monitoring food additives and other toxic contaminants. Sample preparation, such as extraction, concentration and isolation of analytes, greatly influences the reliable and accurate analysis of food. Solid-phase microextraction (SPME) is a new sample preparation technique using a fused-silica fiber that is coated on the outside with an appropriate stationary phase. Analyte in the sample is directly extracted to the fiber coating. The SPME technique can be used routinely in combination with gas chromatography (GC), GC–mass spectrometry (GC–MS), high-performance liquid chromatography (HPLC) or LC–MS. Furthermore, another SPME technique known as in-tube SPME has also been developed for combination with LC or LC–MS using an open tubular fused-silica capillary column as an SPME device instead of SPME fiber. These methods using SPME techniques save preparation time, solvent purchase and disposal costs, and can improve the detection limits. This review summarizes the SPME techniques for coupling with various analytical instruments and the applications of these techniques to food analysis.
Article
There are many different species of cannabis plants, but their psychoactive properties mainly depend on the concentration of tetrahydrocannabinol (THC), which may vary according to genetic factors and environmental influences. On the basis of the THC content all cannabis plants are divided into fibre-type and drug-type plants. The fibre-type plant does not exceed 0.4 per cent of THC while the drug-type plant usually contains up to 5 per cent of THC, though higher percentages (up to 10 per cent) have been reported. A study of the characteristics of cannabis seeds and the influence of environmental conditions on the content of THC in cannabis plants grown in northern, southern and insular Italy has shown that the fibre-type plants contain mean values of THC in a range from 0.058 to 0.299 per cent. The content of THC in the drug-type plants grown in Sicily and Tuscany ranged from 0.82 to 1.31 per cent +/- 0.49 per cent. In 1984, the Commission of the European Communities prepared a regulation to prevent diffusion of the drug-type cannabis, providing that raw material could not be imported if its THC content exceeded 0.5 per cent from 1984 to 1987 and after that period the maximum limit would be set up to 0.3 per cent.
Article
Solid-phase microextraction (SPME), with the poly(dimethylsiloxane)-coated silica fiber suspended and equilibrated in the headspace, has been applied to the capillary gas chromatographic (GC) analysis of 33 halogenated volatile contaminants in model aqueous solutions and in foods. With electrolytic conductivity detection, the limits of detection in water ranged from 1.5 micrograms/kg for vinyl chloride to < or = 0.005 microgram/kg for the tri- to hexachlorobenzenes. Headspace SPME-GC shows a much greater response for the less volatile analytes than those of greater volatility, a procedure complementing headspace GC with gas sampling. In model systems or foods, increasing lipid material decreased the headspace extraction. With 50 mg of lipid, the headspace extraction decreased about 50% for analytes with LODs about 0.1 microgram/kg and by > or = 99.5% for the above chlorobenzenes. Standard addition was used to analyze a variety of beverages and dry foods and to determine the analyte partitions.
Article
Solid-phase microextraction (SPME) was used to determine methylcyclopentadienyl manganese tricarbonyl (MMT), a gasoline antiknock additive, in beverages. MMT levels in beverages exposed to gasoline vapours for 1 h ranged from 0.62 to 2.84 ng/ml and continued to increase up to 16 h. A capillary gas chromatograph (GC) was coupled to an atomic absorption spectrometer (AAS) for element specific detection. The limit of detection was 4 pg MMT. Method limits of detection for MMT varied from 0.4 pg to 260 pg/ml depending on the beverage.
Article
We describe 4 patients who suffered gastrointestinal disorders and psychological effects after eating salad prepared with hemp seed oil. The concentration of tetrahydrocannabinol (THC) in this oil far exceeded the recommended tolerance dose. Our observations prompted the Swiss Federal Office of Public Health to publish warnings in the press concerning consumption of this oil. We describe the symptoms of orally ingested THC and point out unresolved problems connected with food containing hemp.
Article
Solid-phase microextraction (SPME) is applied to the determination of cannabidiol, delta 8-tetrahydrocannabinol (delta 8-THC), delta 9-tetrahydrocannabinol (delta 9-THC), and cannabinol in pure water and human saliva. The inherent extraction behavior of the cannabinoids in pure water is evaluated along with optimization of the method in human saliva. The commercially available poly(dimethylsiloxane) (PDMS) SPME fibers were found to be the best class for the cannabinoid analysis. Partition coefficients were found to be extremely large for all of the cannabinoids (log K > 4.0). Equilibrium times for the 7- and 30-micron PDMS fibers were 50 and 240 min, respectively. A shorter extraction time of 10 min with the 30-micron PDMS fiber may be used for multiple extractions from the same vial, thus conserving the sample necessary for analysis and speeding up the total analysis time. Recoveries for the cannabinoids in saliva, relative to pure water, were dramatically improved by a method developed in our laboratory involving addition of glacial acetic acid to the sample vial prior to performing SPME. Using this method, recoveries relative to SPME in pure water ranged from 21 to 47% depending on the cannabinoid. The linear range for spiked saliva samples was established at 5-500 ng/mL (r2 > 0.994) with precisions between 11 and 20% RSD. The ultimate level of detection by SPME for the cannabinoids in saliva was 1.0 ng/mL, with signal-to-noise values of > or = 12. A saliva sample collected 30 min after marijuana smoking was subject to SPME and traditional liquid-liquid extraction analysis. Internal standard quantitation results for delta 9-THC by both methods yielded comparable results, indicating that the SPME method of analysis is highly accurate and precise. The level of delta 9-THC by SPME was found to be 9.54 ng/mL for the saliva sample.
Article
Headspace solid phase microextraction (HS-SPME) has advantages of high purity of the extract, avoidance of organic solvents and simple technical manipulation and can be used in combination with gas chromatography-mass spectrometry (GC-MS) in the hair analysis of a number of drugs. HS-SPME coupled with the hydrolysis of the hair matrix by 4% sodium hydroxide in the presence of excess sodium sulphate and of a suitable internal standard proved to be a convenient one-step method for the measurement of many lipophilic basic drugs such as nicotine, amphetamine derivatives, local anaesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines. Detection limits were between 0.05 and 1.0 ng/mg. From spiked 10-mg hair samples absolute recoveries between 0.04 and 5.7% were found. These recoveries decreased considerably if larger sample amounts were used, perhaps due to increased drug solubility in the aqueous phase or to elevated viscosity in the presence of dissolved hair proteins. Because of the phenolic hydroxyl group a change of pH after alkaline hair digestion (by adding excess orthophosphoric acid) was necessary for the detection of delta 9-tetrahydrocannabinol (delta 9-THC), cannabinol (CBN) and cannabidiol (CBD) by HS-SPME. Nevertheless, the detection limits were such that only CBN could be detected in hair of a consumer. Clomethiazole, a compound hydrolysed in alkali, was measured by HS-SPME after extraction with aqueous buffer. The detection limit was 0.5 ng/mg. Cocaine could not be detected by HS-SPME. The application of HS-SPME to hair samples from several forensic and clinical cases is described.
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
There has been a recent and significant increase in the use and availability of hemp seed oil products. These products are being marketed as a healthy source of essential omega fatty acids when taken orally. Although the health aspects of these oils is open to debate, the probability that oils derived from the hemp seed will contain delta9-tetrahyrdocannabinol (THC) is noteworthy. Recent additions to the literature cite a number of studies illustrating that the ingestion of these products results in urinary levels of the THC metabolite, delta9-tetrahyrdocannabinol carboxylic acid (THCA), well above the administrative cutoff (50 ng/mL) used during random drug screens. Testing performed by our laboratory found that the concentration of THC in hemp oil products has been reduced considerably since the publication of earlier studies. The purpose of this study is to quantitate the THC levels in commercially available hemp oils and to administer those oils tested to THC-free volunteers to determine urine metabolite levels following several 15-g doses. Two extraction protocols were evaluated for removing THC from the oil matrix: a single step liquid-liquid extraction was compared to a two-phase process using both liquid-liquid and solid-phase techniques. Gas chromatography-mass spectrometry was used to determine THC levels in several products: four from Spectrum Essentials (3 bottled oils and 1-g capsules), two from Health from the Sun (1-g capsules and bottled oil) oils, and single samples of both Hempstead and Hempola hemp oils. These hemp oil products contained THC concentrations of 36.0, 36.4, 117.5, 79.5, 48.6, 45.7, 21.0, and 11.5 mg/g, respectively. The Abbott AxSYM FPIA and Roche On-Line KIMS immunoassays were used to screen the urine samples, and GC-MS was used to determine the amount of THC in each oil as well as confirm and quantitate THCA in the urine of study participants immediately before and 6 h after each dose. Peak THCA levels in the participants' urine ranged from 1 to 49 ng/mL. All volunteers were below positive screen and confirmation cutoffs within 48 h after cessation of ingestion.
Article
Foods containing seeds or oil of the hemp plant (Cannabis sativa L.) are increasingly found in retail stores in the U.S. The presence of delta9-tetrahydrocannabinol (THC) in these foods has raised concern over their impact on the results of workplace drug tests for marijuana. Previous studies have shown that eating hemp foods can cause screening and confirmed positive results in urine specimens. This study evaluated the impact of extended daily ingestion of THC via hemp oil on urine levels of its metabolite 11-nor-9-carboxy-delta9-tetrahydrocannabinol (THC-COOH) for four distinct daily THC doses. Doses were representative of THC levels now commonly found in hemp seed products and a range of conceivable daily consumption rates. Fifteen THC-naïve adults ingested, over four successive 10-day periods, single daily THC doses ranging from 0.09 to 0.6 mg. Subjects self-administered THC in 15-mL aliquots (20 mL for the 0.6-mg dose) of four different blends of hemp and canola oils. Urine specimens were collected prior to the first ingestion of oil, on days 9 and 10 of each of the four study periods, and 1 and 3 days after the last ingestion. All specimens were screened for cannabinoids by radioimmunoassay (Immunalysis Direct RIA Kit), confirmed for THC-COOH by gas chromatography-mass spectrometry (GC-MS), and analyzed for creatinine to identify dilute specimens. None of the subjects who ingested daily doses of 0.45 mg of THC screened positive at the 50-ng/mL cutoff. At a daily THC dose of 0.6 mg, one specimen screened positive. The highest THC-COOH level found by GC-MS in any of the specimens was 5.2 ng/mL, well below the 15-ng/mL confirmation cutoff used in federal drug testing programs. A THC intake of 0.6 mg/day is equivalent to the consumption of approximately 125 mL of hemp oil containing 5 microg/g of THC or 300 g of hulled seeds at 2 microg/g. These THC concentrations are now typical in Canadian hemp seed products. Based on our findings, these concentrations appear to be sufficiently low to prevent confirmed positives from the extended and extensive consumption of hemp foods.
Article
This paper describes a fully automated procedure using alkaline hydrolysis and headspace solid-phase microextraction (HS-SPME) followed by on-fiber derivatization and gas chromatographic-mass spectrometric (GC-MS) detection of cannabinoids in human hair samples. Ten milligrams of hair was washed with deionized water, petroleum ether, and dichloromethane. After the addition of deuterated internal standards the sample was hydrolyzed with sodium hydroxide and directly submitted to HS-SPME. After absorption of analytes for an on-fiber derivatization procedure the fiber was directly placed into the headspace of a second vial containing N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) before GC-MS analysis. The limit of detection was 0.05 ng/mg for delta9-tetrahydrocannabinol (THC), 0.08 ng/mg for cannabidiol (CBD), and 0.14 ng/mg for cannabinol (CBN). Absolute recoveries were in the range between 0.3 and 7.5%. Linearity was proved over a range from 0.1 to 20 ng/mg with coefficients of correlation from 0.998 to 0.999. Validation of the whole procedure revealed excellent results. In comparison with conventional methods of hair analysis this automated HS-SPME-GC-MS procedure is substantially faster. It is easy to perform without use of solvents and with minimal sample quantities, but with the same degree of sensitivity and reproducibility. The applicability was demonstrated by the analysis of 25 hair samples from several forensic cases. The following concentration ranges were determined: THC 0.29-2.20 (mean 1.7) ng/mg, CBN 0.55-4.54 (mean 1.2) ng/mg, and CBD 0.53-18.36 (mean 1.3) ng/mg. 11-nor-Delta9-tetrahydrocannabinol-9-carboxylic acid could not be detected with this method.
Article
A new method combination, headspace solid-phase dynamic extraction coupled with gas chromatography/tandem mass spectrometry (HS-SPDE/GC/MS/MS), is introduced to determine drugs of abuse in hair samples. This highly automated procedure utilizes SPDE for pre-concentration and on-coating derivatization as well as GC and triple quadrupole MS/MS for selective and sensitive detection. All these steps, apart from washing and cutting of the hair samples, are performed without manual intervention on a robot-like autosampler.SPDE is a solventless extraction technique related to solid-phase microextraction (SPME). The analytes are absorbed from the sample headspace directly into a hollow needle with an internal coating of polydimethylsiloxane by repeated aspirate/dispense cycles.The HS-SPDE/GC/MS/MS procedure was applied to the analysis of methadone, the trimethylsilyl derivatives of cannabinoids and the trifluoroacetyl derivatives of amphetamines and designer drugs. The method was shown to be sensitive with detection limits between 6 and 52 pg/mg hair matrix and precision between 0.4 and 7.8% by the use of an internal standard technique. Linearity was obtained from 0.1-20 ng/mg with coefficients of correlation between 0.995 and 0.999. Compared with conventional methods of hair analysis, HS-SPDE/GC/MS/MS is easier to use, substantially faster, with the degree of sensitivity and reproducibility demanded in clinical and forensic toxicology. The main advantage of the SPDE technique in relation to SPME is the robustness of the capillary.
Article
This article describes a fully automated procedure for detecting cannabinoids in human hair samples. The procedure uses alkaline hydrolysis and headspace solid-phase dynamic extraction (HS-SPDE), followed by on-coating derivatization and gas chromatography-mass spectrometry (GC-MS). SPDE is a further development of solid-phase microextraction (SPME), based on an inside needle capillary absorption trap. It uses a hollow needle with an internal coating of polydimethylsiloxane as extraction and pre-concentration medium. Ten mg of hair were washed with deionised water, petroleum ether and dichloromethane. After adding deuterated internal standards, the sample was hydrolyzed with sodium hydroxide and directly submitted to HS-SPDE. After absorption of analytes for an on-coating derivatization procedure, the SPDE-needle was directly placed into the headspace of a second vial containing N-methyl-N-trimethylsilyl-trifluoroacetamide before GC-MS analysis. The limit of detection was 0.14 ng/mg for Delta(9)-tetrahydrocannabinol, 0.09 ng/mg for cannabidiol, and 0.12ng/mg for cannabinol. Absolute recoveries were in the range of 0.6 to 8.4%. Linearity was verified over a range from 0.2 to 20 ng/mg, with coefficients of correlation between 0.998 and 0.999. Intra- and inter-day precision were determined at two different concentrations and resulted in ranges between 2.3 and 6.0% (intra-day) and 3.3 and 7.6% (inter-day). Compared with conventional methods of hair analysis, this automated HS-SPDE-GC-MS procedure is substantially faster. It is easy to perform without using solvents and with minimal sample quantities, and it yields the same sensitivity and reproducibility. Compared to SPME, we found a higher extraction rate, coupled with a faster automated operation and greater stability of the device.
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