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Multidimensional analysis of cannabis volatile constituents: Identification of 5,5-dimethyl-1-vinylbicyclo[2.1.1]hexane as a volatile marker of hashish, the resin of Cannabis sativa L.
Abstract
The volatile constituents of drug samples derived from Cannabis sativa L. were investigated by means of headspace solid phase microextraction (HS-SPME) and gas chromatography techniques (GC-MS, GC×GC-MS). Samples of cannabis herb and hashish showed clear differences in their volatile chemical profiles, mostly resulting from photo-oxidation processes occurring during the transformation of fresh cannabis herb into hashish. Most unexpectedly, we could demonstrate hashish samples as containing remarkable amounts of a rare and unusual monoterpene - 5,5-dimethyl-1-vinylbicyclo[2.1.1]hexane - among the volatile compounds detected in their headspaces. We gave evidence for the formation of this compound from the light induced rearrangement of β-myrcene during the manufacture of hashish. In view of its high abundance among volatile constituents of cannabis resin and its scarce occurrence in other natural volatile extracts, we propose to rename this specific monoterpene hashishene.
Copyright © 2014 Elsevier B.V. All rights reserved.
... When administered orally, a single dose of βmyrcene has been shown to extend the duration of pentobarbital-induced sleep when administered 60 min before a barbiturate [98]. Additionally, β-myrcene undergoes photooxidation to form "hashishene", a compound notable for its high concentration in hashish [99]. ...
... When administered orally, a single dose of β-myrcene has been shown to extend the duration of pentobarbital-induced sleep when administered 60 min before a barbiturate [98]. Additionally, β-myrcene undergoes photo-oxidation to form "hashishene", a compound notable for its high concentration in hashish [99]. Linalool (20) Anxiolytic and antidepressant Linalool, a major compound of lavender essential oil, is traditionally used and has been approved by the EMA as an herbal medicinal product for alleviating mild symptoms of mental stress and exhaustion and aiding sleep [74]. ...
... When administered orally, a single d myrcene has been shown to extend the durati pentobarbital-induced sleep when administered 60 m barbiturate [98]. Additionally, β-myrcene undergo oxidation to form "hashishene", a compound notable concentration in hashish [99]. ...
This study explores the complementary or synergistic effects of medicinal cannabis constituents, particularly terpenes, concerning their therapeutic potential, known as the entourage effect. A systematic review of the literature on cannabis “entourage effects” was conducted using the PRISMA model. Two research questions directed the review: (1) What are the physiological effects of terpenes and terpenoids found in cannabis? (2) What are the proven “entourage effects” of terpenes in cannabis? The initial approach involved an exploratory search in electronic databases using predefined keywords and Boolean phrases across PubMed/MEDLINE, Web of Science, and EBSCO databases using Medical Subject Headings (MeSH). Analysis of published studies shows no evidence of neuroprotective or anti-aggregatory effects of α-pinene and β-pinene against β-amyloid-mediated toxicity; however, modest lipid peroxidation inhibition by α-pinene, β pinene, and terpinolene may contribute to the multifaceted neuroprotection properties of these C. sativa L. prevalent monoterpenes and the triterpene friedelin. Myrcene demonstrated anti-inflammatory proprieties topically; however, in combination with CBD, it did not show significant additional differences. Exploratory evidence suggests various therapeutic benefits of terpenes, such as myrcene for relaxation; linalool as a sleep aid and to relieve exhaustion and mental stress; D-limonene as an analgesic; caryophyllene for cold tolerance and analgesia; valencene for cartilage protection; borneol for antinociceptive and anticonvulsant potential; and eucalyptol for muscle pain. While exploratory research suggests terpenes as influencers in the therapeutic benefits of cannabinoids, the potential for synergistic or additive enhancement of cannabinoid efficacy by terpenes remains unproven. Further clinical trials are needed to confirm any terpenes “entourage effects.”
... The use of GC × GC in detecting a higher number of compounds was also demonstrated by Marchini and colleagues. They were able to detect three times more compounds in the extracts of cannabis herb and hashish as compared to the conventional GC-MS [26]. ...
... We also detected 32 more compounds in our samples compared to a GC × GC study on cannabis and hashish by Marchini and colleagues [26]. However, the number of volatile compounds in cannabis and cannabis derivatives depends on various factors, such as extraction method, chromatographic method, and type of cannabis species and strain used, as well as the nature of the cannabis flowers used, i.e., fresh or dried. ...
... However, the number of volatile compounds in cannabis and cannabis derivatives depends on various factors, such as extraction method, chromatographic method, and type of cannabis species and strain used, as well as the nature of the cannabis flowers used, i.e., fresh or dried. Marchini and colleagues found 16 more monoterpenoids than in our study as they used hashish, which contains a higher number of oxygenated terpenes than cannabis herb [26]. ...
Cannabis contains a wide range of terpenes and terpenoids that are mainly responsible for their distinctive aroma and flavor. These compounds have also demonstrated therapeutic effects either alone and/or as synergistic compounds with other terpenes, terpenoids, and/or cannabinoids. Several studies have attempted to fully characterize terpenes and terpenoids in cannabis; however, most of these studies used one-dimensional gas chromatography, which often results in the co-elution of the compounds. In the present study, we analyzed terpenes and terpenoids in the dried flowers of six cannabis strains using a two-dimensional gas chromatograph time-of-flight mass spectrometer (GC × GC-TOFMS). A total of 146 terpenes and terpenoids were detected across all six cannabis strains with an enhanced separation of 16 terpenes and terpenoids in the second dimension. Additionally, we achieved enhanced separation of four terpenes and terpenoids from a standard mixture in the second dimension. Chemical differences were observed in the number and relative abundance of monoterpenes, monoterpenoids, sesquiterpenes, and sesquiterpenoids in all six strains. We were also able to identify four new terpenoids in cannabis, which are reported here for the first time.
... 6 Among the vast number of terpenoids in Cannabis, only approximately 20 to 30 terpenoids have been reported and used for chemotyping of Cannabis cultivars. [7][8][9][10][11][12][13][14][15] Hundreds of additional terpenoids and other volatiles have been recently identified in Cannabis, [16][17][18] but their absolute concentrations have never been determined using a validated analytical method. ...
... 28 Also, several HS-SPME-GC-MS methods have been recently reported for comprehensive profiling of terpenoids in Cannabis. [16][17][18] HS-SPME, however, is considered less robust and precise than SHS, as terpenoid concentrations injected may be affected by the nature of the fiber selected for the extraction. 17 Also, SPME methods are often applied for improving signal intensity in diluted samples. ...
... Due to their unsaturated nature, terpenoids are highly prone to photo-oxidation. 17,18 Much like in triglyceride oils and other polyunsaturated molecules, photooxidation leads to the formation of allylic hydroperoxides. 17,18 These are highly reactive species, that may undergo further decomposition into oxygenated products including alcohols, ketones, and aldehydes. ...
The therapeutic effect of Cannabis largely depends on the content of its pharmacologically active secondary metabolites, mainly phytocannabinoids, flavonoids and terpenoids. Recent studies suggest of therapeutic effects of specific terpenoids, as well as synergistic effects with other active compounds in the plant. Although Cannabis contains an overwhelming milieu of terpenoids, only a limited number are currently reported and used for metabolic analysis of Cannabis chemovars. In this study, we report the development and validation of a method for simultaneous quantification of 93 terpenoids in Cannabis air-dried-inflorescences and extracts. This method employs the full evaporation technique via a static headspace sampler, followed by gas chromatography–mass spectrometry (SHS-GC–MS/MS). In the validation process, spiked terpenoids were quantified with acceptable repeatability, reproducibility, sensitivity and accuracy. Three medical Cannabis chemovars were used to study the effect of sample preparation and extraction methods on terpenoid profiles. This method was further ap-plied for studying the terpenoid profiles of sixteen different chemovars acquired at different dates. Our results demonstrate that sample preparation methods may significantly impact the chemical fingerprint compared to the non-treated Cannabis. This emphasizes the importance of performing SHS extraction in order to study the natural terpenoid contents of che-movars. We also concluded that most inflorescences expressed relatively unique terpenoid profiles for the most pronounced terpenoids, even when sampled at different dates, although absolute concentrations may vary due to aging. The suggested method offer an ideal tool for terpenoid profiling of Cannabis and set the scene for more comprehensive works in the fu-ture.
... As regards the other compounds present in hemp, terpenes are responsible for the characteristic aroma of the plant. The main volatiles detected in the aerial parts of the plant include both monoand sesquiterpenes [14,15,17,[27][28][29], with β-myrcene and β-caryophyllene as the most representative compounds (Figure 3), respectively. As regards monoterpenes, β-myrcene is known to possess anti-inflammatory, analgesic, and anxiolytic properties [17]. ...
... As regards the other compounds present in hemp, terpenes are responsible for the characteristic aroma of the plant. The main volatiles detected in the aerial parts of the plant include both monoand sesquiterpenes [14,15,17,[27][28][29], with β-myrcene and β-caryophyllene as the most representative compounds (Figure 3), respectively. As regards monoterpenes, β-myrcene is known to possess antiinflammatory, analgesic, and anxiolytic properties [17]. ...
... HS-SPME is being increasingly selected as the technique of choice in the ambit of the extraction of volatile compounds in the field of natural product research [14,15,27,28]. In this study, a new HS-SPME procedure was optimised in order to achieve a reliable characterisation of the volatiles from hemp inflorescences. ...
Cannabis sativa L. is a dioecious plant belonging to the Cannabaceae family. The main phytochemicals that are found in this plant are represented by cannabinoids, flavones, and terpenes. Some biological activities of cannabinoids are known to be enhanced by the presence of terpenes and flavonoids in the extracts, due to a synergistic action. In the light of all the above, the present study was aimed at the multi-component analysis of the bioactive compounds present in fibre-type C. sativa (hemp) inflorescences of different varieties by means of innovative HPLC and GC methods. In particular, the profiling of non-psychoactive cannabinoids was carried out by means of HPLC-UV/DAD, ESI-MS, and MS². The content of prenylated flavones in hemp extracts, including cannflavins A and B, was also evaluated by HPLC. The study on Cannabis volatile compounds was performed by developing a new method based on headspace solid-phase microextraction (HS-SPME) coupled with GC-MS and GC-FID. Cannabidiolic acid (CBDA) and cannabidiol (CBD) were found to be the most abundant cannabinoids in the hemp samples analysed, while β-myrcene and β-caryophyllene were the major terpenes. As regards flavonoids, cannflavin A was observed to be the main compound in almost all the samples. The methods developed in this work are suitable for the comprehensive chemical analysis of both hemp plant material and related pharmaceutical or nutraceutical products in order to ensure their quality, efficacy, and safety.
... Monitoring illicit substances such as illegal drugs, explosives and chemical warfare agents (CWAs) is of particular importance for public security in strategic locations, but also for the identification of their origin/history which can be used as evidence in court [4]. The low detection limits demanded usually result in targeted approaches [30], which would neglect derivatives, transformation products or relevant contaminants. Due to its high separation By applying Fisher ratios on the data set to create two-dimensional Fisher ratio plots, 221 peaks of interest were found to have the maximum potential for class differentiation. ...
... capacity, GC Â GC-MS allows for both targeted and untargeted analysis and is therefore of particular forensic interest for the identification, characterization and classification of material containing illegal substances [4]. Recently, GC Â GC-MS has been applied to the study of nerve agents [31], explosives [32,33] and illicit drugs such as cannabis [30,34], heroin [35], and steroids [36,37]. To efficiently separate security-relevant trace components from matrix interferences in GC Â GC, the trend is towards a normal phase column set. ...
... Applying a design of experiments (DoE) optimized SPME-GC Â GC-MS method, Marchini et al. [30] differentiated the volatile chemical profiles of cannabis derived products. Compared to 1 Dprofiles, compound identification was greatly improved. ...
Comprehensive two-dimensional gas chromatography (GC×GC) has become accepted as one of the most powerful separation techniques in several application areas. In forensic investigations, however, it has not yet been entirely established due to limitations regarding standardized methodology, data interpretation and consistency of results. Nevertheless, GC×GC allows for target analysis, compound class analysis and chemical fingerprinting of samples and is therefore increasingly applied in forensic analytics. In this review, recent and significant advances in GC×GC for application to forensic studies including human scent, arson investigations, security-relevant substances and environmental forensics are discussed. The discussion includes a brief overview of the latest trends and evolutions with regard to the various forensic applications and data evaluation as well as limitations. This leads to the conclusion that the full potential of the comprehensive data sets can only be achieved by implementing standardized analysis and data processing methods.
... Samples 5, 12 and 14 contained a distinct number of various terpenes, although in far lower concentration compared to sample Oil_6. Apparently, these formulations were obtained by an extraction process able to preserve naturally occurring terpenes profile from initial Cannabis sativa plants, as their terpene profile is in accordance with those already published in literature [12,[21][22][23][24]35]. Similarly, Bedrolite oil extract (Oil_1) contains various terpene structures, reflecting the initial plant profiling. ...
... This remains unclear considering that the two pinenes are quite balanced within the different Cannabis varieties representing around the 10% of the terpenes group and not exceeding 15-20% 20. The occurrence of α-terpinolene in all samples (except Oil_2 and Oil_13), might be important as this compound was suggested as a genetic marker for distinguishing two important gene pools for breeding low-THC varieties [35,36]. Sample Oil_14 was particularly rich in trans-caryophyllene followed by α-humulene. ...
... This remains unclear considering that the two pinenes are quite balanced within the different Cannabis varieties representing around the 10% of the terpenes group and not exceeding 15-20% [20]. The occurrence of α-terpinolene in all samples (except Oil_2 and Oil_13), might be important as this compound was suggested as a genetic marker for distinguishing two important gene pools for breeding low-THC varieties [35,36]. Sample Oil_14 was particularly rich in trans-caryophyllene followed by α-humulene. ...
Cannabidiol (CBD)-based oil preparations are becoming extremely popular, as CBD has been shown to have beneficial effects on human health. CBD-based oil preparations are not unambiguously regulated under the European legislation, as CBD is not considered as a controlled substance. This means that companies can produce and distribute CBD products derived from non-psychoactive hemp varieties, providing an easy access to this extremely advantageous cannabinoid. This leaves consumers with no legal quality guarantees. The objective of this project was to assess the quality of 14 CBD oils commercially available in European countries. An in-depth chemical profiling of cannabinoids, terpenes and oxidation products was conducted by means of GC-MS and HPLC-Q-Exactive-Orbitrap-MS in order to improve knowledge regarding the characteristics of CBD oils. Nine out of the 14 samples studied had concentrations that differed notably from the declared amount, while the remaining five preserved CBD within optimal limits. Our results highlighted a wide variability in cannabinoids profile that justifies the need for strict and standardized regulations. In addition, the terpenes fingerprint may serve as an indicator of the quality of hemp varieties, while the lipid oxidation products profile could contribute in evaluation of the stability of the oil used as milieu for CBD rich extracts.
... Scarce data are available about the application of HS in the analysis of terpenes and in general of the volatile profile from medical cannabis varieties. Currently, Marchini et al. applied SPME to characterise the terpene profile from hashish, the exudate resin of C. sativa L., seeking for potential volatile markers to distinguish the resin from plant material [17]. ...
... In addition, terpenes were extracted using methanol as extraction solvent and then quantified using a calibration curve constructed on the basis of one terpene only. This approach is usually limitative, as the polarity of the solvent could dramatically influence the obtained terpene profile, leading to the underestimation of the complex mixture of secondary metabolites emitted by plants [17]. ...
... Most of these oxygenated compounds corresponded to secondary photo-oxidation's products of the initial terpenes. In presence of light and singlet oxygen, terpenes are known to undergo photo-oxidation leading to the formation of allylic hydroperoxides [17]. Since hydroperoxides are generally unstable, they will generate the corresponding alcohols that may be further oxidized to the corresponding aldehydes and ketones [17], e.g. the degradation of limonene that under photo-oxidation leads to the formation of trans-and cismetha-2,8-dien-1-ol and trans-and cis-carveol. ...
There are at least 554 identified compounds in C. sativa L., among them 113 phytocannabinoids and 120 terpenes. Phytocomplex composition differences between the pharmaceutical properties of different medical cannabis chemotype have been attributed to strict interactions, defined as 'entourage effect', between cannabinoids and terpenes as a result of synergic action. The chemical complexity of its bioactive constituents highlight the need for standardised and well-defined analytical approaches able to characterise the plant chemotype, the herbal drug quality as well as to monitor the quality of pharmaceutical cannabis extracts and preparations. Hence, in the first part of this study an analytical procedures involving the combination of headspace-solid-phase microextraction (HS-SPME) coupled to GC-MS and High Resolution Mass-Spectrometry LC-HRMS (Orbitrap®) were set up, validated and applied for the in-depth profiling and fingerprinting of cannabinoids and terpenes in two authorised medical grade varieties of Cannabis sativa L. inflorescences (Bedrocan® and Bediol®) and in obtained macerated oils. To better understand the trend of all volatile compounds and cannabinoids during oil storage a new procedure for cannabis macerated oil preparation without any thermal step was tested and compared with the existing conventional methods to assess the potentially detrimental effect of heating on overall product quality.
... 37 In the case of cannabis, a photolytic degradation of -myrcene to hashishene was observed. 40 Furthermore, the enantiomeric ratio can be altered, which is of significance for, for example, authenticity control and aging, due to enantioselective terpene synthesis in plants. 29,[41][42][43] Changes in the enantiomeric ratio of terpenes can affect their pharmacological activity. ...
... The formation of hashishene from myrcene, as suggested by Marchini et al., was not confirmed in control samples. 40 Mehdizadeh et al. conducted a study on the stability of Cuminum cyminium L. under different storage temperatures. Similar to our results, they also observed a moderate decrease of αand β-pinene and a rapid decline of myrcene at room temperature. ...
Objective
Cannabis sativa L. is renowned for its medicinal and recreational uses. With the increasing global legalization of C. sativa L.‐based products for medicinal purposes, there is a growing need for well‐characterized products. While the stability of cannabinoids such as tetrahydrocannabinol and cannabidiol is well understood, information on the chemical and enantiomeric stability of terpenes remains scarce. This is despite the fact that terpenes are also thought to have pharmacological activity and may contribute to the overall effect of C. sativa L.
Methods
To address these challenges, four analytical methods based on chiral, polar, and apolar gas chromatographic separation combined with either MS or FID detection were developed and validated. These methods successfully separated and quantified a total of 29 terpenes, including 13 enantiomers and 5 diastereomers specific to C. sativa L. Furthermore, terpenes and authentic C. sativa L. flowers and extracts were subjected to UV and heat treatments to observe potential degradation reactions over time.
Results
Each terpene generates a unique pattern of degradation products resulting in a diverse array of oxidation and cyclization products. P‐cymene was identified as a major product of terpene aging. Notably, no enantiomeric conversion was detected, suggesting that the formation of (−)‐α‐pinene in cannabis extracts, for example, originates from other terpenes.
Conclusion
Terpenes have different degradation rates, even though they are structurally similar. In addition, cultivar‐ and growth‐condition‐specific enantiomeric ratios were observed in C. sativa L., confirming that enantiomer production is species‐specific and has to be considered for therapeutical applications.
... 11,24 To overcome the inaccuracies of the indica/sativa binary classification and better categorize cannabis varieties based on their psychoactive and aroma characteristics, terpenes have emerged as a prominent focus of research. 17,19,25,26 Terpenes, diverse organic compounds found in various plants, including cannabis, contribute to the distinct aromas and flavors of different varieties. 27 More importantly, terpenes are believed to play a significant role in the psychoactive and medicinal properties of cannabis, with specific terpenes potentially correlating with distinct psychoactive effects. ...
... While previous studies have detailed a number of these compounds, few have described them in the context of how they affect the aromatic properties of specific varieties. 18,19 Many modern cannabis varieties are often described as exotic, which we define as varieties that are unusually sweet or savory. The former often have aroma descriptors such as sweet or fruity, while the latter include chemical or savory. ...
Cannabis sativa L. produces a wide variety of volatile secondary metabolites that contribute to its unique aroma. The major volatile constituents include monoterpenes, sesquiterpenes, and their oxygenated derivates. In particular, the compounds ß-myrcene, D-(+)-limonene, ß-caryophyllene, and terpinolene are often found in greatest amounts, which has led to their use in chemotaxonomic classification schemes and legal Cannabis sativa L. product labeling. While these compounds contribute to the characteristic aroma of Cannabis sativa L. and may help differentiate varieties on a broad level, their importance in producing specific aromas is not well understood. Here, we show that across Cannabis sativa L. varieties with divergent aromas, terpene expression remains remarkably similar, indicating their benign contribution to these unique, specific scents. Instead, we found that many minor, nonterpenoid compounds correlate strongly with nonprototypical sweet or savory aromas produced by Cannabis sativa L. Coupling sensory studies to our chemical analysis, we derive correlations between groups of compounds, or in some cases, individual compounds, that produce many of these diverse scents. In particular, we identified a new class of volatile sulfur compounds (VSCs) containing the 3-mercaptohexyl functional group responsible for the distinct citrus aromas in certain varieties and skatole (3-methylindole) as the key source of the chemical aroma in others. Our results provide not only a rich understanding of the chemistry of Cannabis sativa L. but also highlight how the importance of terpenes in the context of the aroma of Cannabis sativa L. has been overemphasized.
... Over 200 aroma compounds have previously been reported in cannabis, highlighting the complexity of its odor. [9][10][11]22 The compounds with the highest concentrations typically include the terpenoids β-myrcene, αand β-pinene, D-limonene, βcaryophyllene, terpinolene, and humulene, which can individually contribute upward of 50% of the aroma concentration. [9][10][11]13,23 The types and relative concentrations of these compounds contribute significantly to the scent of cannabis, which is becoming increasingly diverse as cultivars are crossbred. ...
... [9][10][11]22 The compounds with the highest concentrations typically include the terpenoids β-myrcene, αand β-pinene, D-limonene, βcaryophyllene, terpinolene, and humulene, which can individually contribute upward of 50% of the aroma concentration. [9][10][11]13,23 The types and relative concentrations of these compounds contribute significantly to the scent of cannabis, which is becoming increasingly diverse as cultivars are crossbred. For instance, OG Kush, a cannabis indica cultivar, possesses a strong, pungent, fuel-like aroma that arises from high concentrations of β-myrcene and β-caryophyllene. ...
Cannabis sativa L. produces over 200 known secondary metabolites that contribute to its distinctive aroma. Studies on compounds traditionally associated with the scent of this plant have focused on those within the terpenoid class. These isoprene-derived compounds are ubiquitous in nature and are the major source of many plant odors. Nonetheless, there is little evidence that they provide the characteristic “skunk-like” aroma of cannabis. To uncover the chemical origins of this scent, we measured the aromatic properties of cannabis flowers and concentrated extracts using comprehensive two-dimensional gas chromatography equipped with time-of-flight mass spectrometry, flame ionization detection, and sulfur chemiluminescence. We discovered a new family of volatile sulfur compounds (VSCs) containing the prenyl (3-methylbut-2-en-1-yl) functional group that is responsible for this scent. In particular, the compound 3-methyl-2-butene-1-thiol was identified as the primary odorant. We then conducted an indoor greenhouse experiment to monitor the evolution of these compounds during the plant’s lifecycle and throughout the curing process. We found that the concentrations of these compounds increase substantially during the last weeks of the flowering stage, reach a maximum during curing, and then drop after just one week of storage. These results shed light on the chemical origins of the characteristic aroma of cannabis and how volatile sulfur compound production evolves during plant growth. Furthermore, the chemical similarity between this new family of VSCs and those found in garlic (allium sativum) suggests an opportunity to also investigate their potential health benefits.
... Like gamma irradiation, UV is not without potential drawbacks as it has been demonstrated that UV light can convert and isomerize terpenes. One example of this is β-myrcene, which can be converted by UV light into hashishene (Figure 6.6), which is not naturally produced by the cannabis plant (Marchini et al. 2014). More work is needed to determine if specific wavelengths of UV light favor this reaction as this has not been studied in the context of remediation. ...
... UV light-induced rearrangement of beta myrcene into hashishene(Marchini et al. 2014). Further research is needed to determine if UV-based remediation techniques can result in chemical transformations that significantly alter the overall terpene profile. ...
The microbial testing of cannabis and cannabis products presents a unique set of challenges. Unlike food testing, cannabis testing has various routes of administration to take into account. Cannabis flowers express high levels of antimicrobial cannabinoids and terpenoids and thus represent a different matrix than traditional foods. It is currently estimated that 50% of cannabis is consumed via vaporizing or smoking oils and flowers while the other half is consumed as Marijuana Infused Products or MIPs which encompass a wide variety of matrices.
In a testing landscape that consistently focuses heavily on chemical analysis, the microbiological testing of cannabis is often overlooked. However, it is truly one of the most important analyses in the context of product safety as the accidental ingestion or inhalation of these contaminants can cause severe illnesses, infections, or worse, death. The present chapter explores the microbial contaminants of interest in cannabis, current testing methodologies, and the challenges that testing laboratories face in this continuously evolving domain. Different perspectives for ensuring product safety are presented in the context of current regulations and their varying approaches. Tactics for the remediation of contaminated product and preventative strategies used by cultivators are also discussed in the context of the existing incongruent patchwork of regulatory framework. Microbial testing acceptance criteria, methods, and recommendations from various standards organizations are presented and efforts towards the standardization and development of reference methods are highlighted.
... Felina 32. Additionally, Marchinini et al. [24] detected abundant quantities of these derivatives both in herb inflorescences and hashish samples using a comprehensive two-dimensional GC system coupled to mass spectrometer (GC × GC-MS). Table 1. ...
... Felina 32. Additionally, Marchinini et al. [24] detected abundant quantities of these derivatives both in herb inflorescences and hashish samples using a comprehensive twodimensional GC system coupled to mass spectrometer (GC × GC-MS). Table 1. ...
A microwave distillation method was optimized for the extraction and isolation of cannabis essential oil from fresh and dried hemp inflorescences. The developed method enabled us to obtain a distilled product rich in terpenes and terpenoid compounds, responsible of the typical and unique smell of the cannabis plant. The distillate from different hemp cultivars, including Kompolti, Futura 75, Carmagnola, Felina 32 and Finola were characterized by using a gas chromatograph equipped with both mass spectrometer and flame ionization detectors. In a single chromatographic run, the identity and absolute amounts of distilled compounds were determined. Peak assignment was established using a reliable approach based on the usage of two identification parameters, named reverse match, and linear retention index filter. Absolute quantification (mg g⁻¹) of the analytes was performed using an internal standard method applying the flame ionization detector (FID) response factors according to each chemical family. An enantio-GC-MS method was also developed in order to evaluate the enantiomeric distribution of chiral compounds, an analytical approach commonly utilized for establishing the authenticity of suspicious samples.
... Measurements of BVOC concentrations in headspace and (illicit) grow rooms have detected and identified many hundreds of BVOCs, often in very low concentrations, of which monoand sesquiterpenes are dominant. These species include: α-pinene, βpinene, β-myrcene, limonene, hashishene, caryophyllene, and humulene (Martyny et al., 2013;Marchini et al., 2014). Hood et al. (Hood et al., 1973) analysed the air above Cannabis spp. ...
... plants and found that the monoterpenes α-pinene, β-pinene, β-myrcene and d-limonene accounted for over 85% of the detected VOCs emitted, with acetone and methanol contributing a further 10%. Marchini et al. (2014) reported the composition of headspace, but not the concentration of each species. Martyny et al. (2013) reported total monoterpene (consisting of αpinene, β-pinene, β-myrcene and d-limonene) concentrations of 50-100 ppbv in the grow rooms of illicit cultivation facilities, suggesting high emissions from growing Cannabis spp.. Due to Cannabis spp. ...
The legal commercialization of Cannabis for recreational and medical use in certain US states has effectively created a new and nearly unregulated cultivation industry. Within the city limits of Denver, Colorado, there are now more than 600 registered Cannabis spp. cultivation facilities (CCFs) for recreational and medical uses, each containing thousands of plants. Ambient measurements collected inside growing operations pre-legalization have found concentrations as high as 50–100 ppbv of terpenes; a group of highly reactive biogenic volatile organic compounds (BVOCs) and known precursors for the formation of ozone and particulate matter (PM). Due to its illicit nature there has been insufficient experimental data produced to determine Cannabis spp. emission rates. This study used, for the first time, an enclosure chamber and live Cannabis spp. plants during a 90-day growing period consisting of four different strains of Cannabis spp.: Critical Mass, Lemon Wheel, Elephant Purple, and Rockstar Kush. These measurements enabled characterization of terpenes and estimates of emission capacity (EC, μgC g⁻¹ hr⁻¹) at standard conditions. During peak growth, the percentages of individual BVOC emissions were dominated by β-myrcene (18–60%), eucalyptol (17–38%), and d-limonene (3–10%) for all strains. Our results showed large variability in the rate and composition of terpene emissions across different strains. For the Critical Mass and Lemon Wheel, the dominant terpenoid was eucalyptol (32% and 38%), and it was β-myrcene (60% and 45%) for the Elephant Purple and Rockstar Kush. Critical Mass produced the highest terpene emission capacity (8.7 μgC g⁻¹ hr⁻¹) and Rockstar Kush the lowest (4.9 μgC g⁻¹ hr⁻¹). With 600 CCFs in Denver, and assuming 10,000 plants per CCF, an emission capacity of 8.7 μgC g⁻¹ hr⁻¹ would more than double the existing rate of BVOC emissions to 520 metric ton year⁻¹. Using Maximum Incremental Reactivity (MIR) values the total ozone formation potential from all these emitted species could produce 2100 metric tons year⁻¹ of ozone, and based on published secondary organic aerosols yields 131 metric tons year⁻¹ of PM. It is likely that the ECs calculated here are lower than those achieved in CCFs where growing conditions are optimized for rapid growth and higher biomass yields. Further studies including a greater number of the 620 available Cannabis spp. strains and a wider range of treatments are needed to generate a representative dataset. Such a dataset could then better enable assessments of the potential impacts of this new industry on indoor and regional air quality.
... When administered orally, a single dose of βmyrcene has been shown to extend the duration of pentobarbital-induced sleep when administered 60 minutes before the barbiturate (Freitas et al., 1993). Additionally, β-myrcene undergoes photo-oxidation to form "hashishene", a compound notable for its high concentration in hashish (Marchini et al., 2014). ...
This study explores the complementary or synergistic effects of medicinal cannabis constituents, particularly terpenes, concerning their therapeutic potential, known as the entourage effect. A systematic review of the literature on cannabis entourage effects was conducted using the PRISMA model. Two research questions conducted the review: (1) What are the Physiological Effects of Terpenes and Terpenoids found in Cannabis? (2) What are the proven Entourage Effects of Terpenes in Cannabis? The initial approach involved an exploratory search in electronic databases using predefined keywords and Boolean phrases across PubMed/MEDLINE, Web of Science, and EBSCO databases, using Medical Subject Headings (MeSH). Analysis of published studies shows no evidence of neuroprotective or anti-aggregatory effects of α-pinene and β-pinene against β-amyloid-mediated toxicity, however, modest lipid peroxidation inhibition by α-pinene, β pinene, and terpinolene may contribute to the multifaceted neuroprotection properties of these C. sativa-prevalent monoterpenes and their triterpene friedelin. Myrcene demonstrated anti-inflammatory proprieties topically, however, in combination with CBD did not show significant additional differences. Exploratory evidence suggests various therapeutic benefits of terpenes, such as myrcene for relaxing; linalool as sleep aid, exhaustion relief and mental stress; D-limonene as an analgesic; caryophyllene for cold tolerance and analgesia; valencene for cartilage protection, borneol for antinociceptive and anticonvulsant potential; and eucalyptol for muscle pain. While exploratory research suggests terpenes as influencers in the therapeutic benefits of cannabinoids, the potential for synergistic or additive enhancement of cannabinoid efficacy by terpenes remains unproven. Further clinical trials are needed to confirm these constituents' individual and combined effects.
... The main illicit cannabis-based products are marijuana (herbal cannabis) and hashish (cannabis resin). The former is obtained mainly from the plant flowering tops together with some leaves [3], while the latter is produced through a relatively long cottage industry process consisting of successively drying, sieving, and finally pressing, or by means of butan hashish oil (BHO) extraction [4]. The illegal market represents one of the main concerns for European authorities, considering that in 2021, the EU Member States declared 202,000 seizures of hashish (equivalent to 816 tons) and 240,000 seizures of marijuana, amounting to 256 tons [1]. ...
... The spontaneous rearrangement of these oxidized products produces alcohols that are often further oxidized to their respective aldehydes and ketones. For example, limonene degrades to transand cis-metha-2,8-dien-1-ol and transand cis-carveol during photooxidation [51,52]. These cannabis oils are activated by heating, which decarboxylates acidic cannabinoids to produce neutral cannabinoids. ...
With the increasing accessibility of cannabis (Cannabis sativa L., also known as marijuana and hemp), its products are being developed as extracts for both recreational and therapeutic use. This has led to increased scrutiny by regulatory bodies, who aim to understand and regulate the complex chemistry of these products to ensure their safety and efficacy. Regulators use targeted analyses to track the concentration of key bioactive metabolites and potentially harmful contaminants, such as metals and other impurities. However, the metabolic complexity of cannabis metabolic pathways requires a more comprehensive approach. A non-targeted metabolomic analysis of cannabis products is necessary to generate data that can be used to determine their authenticity and efficacy. An authentomics approach, which involves combining the non-targeted analysis of new samples with big data comparisons to authenticated historic datasets, provides a robust method for verifying the quality of cannabis products. To meet International Organization for Standardization (ISO) standards, it is necessary to implement the authentomics platform technology and build an integrated database of cannabis analytical results. This study is the first to review the topic of the authentomics of cannabis and its potential to meet ISO standards.
... Nowadays, a wide variety of HS-SPME applications are reported, covering di-verse areas such as industry, pharmacy, food, etc. Among them, this method has gained a special importance in the analysis of terpenes and terpenoids in different Cannabis products [14][15][16][17][18] . Unlike cannabinoids, which are practically hemp exclusive, terpenes can be found in a wide variety of aromatic plants at diverse concentration levels and are responsible for its aroma and flavor. ...
Terpenes and terpenoids are the principal responsible for the aroma of Cannabis, playing an important role in the interaction with the environment. Analytical determination of these compounds can be done by headspace coupled to solid phase micro-extraction (HS-SPME) and then injected in a gas chromatograph. In the present study, we determined distribution constants between gas and polydimetylsiloxane (PDMS), a conventional SPME liquid phase, at three temperatures between 303.15 and 343.15 K for major Cannabis terpenes and terpenoids employing a method based in gas chromatography using four capillary columns for monoterpenes and five columns for sesquiterpenes. In addition, van't Hoff regressions (logKfg vs T-1) were obtained in order to estimate logKfg at 298.15 K aiming to compare with bibliographic values (experimental or estimated ones). An excellent agreement was found between them. The method, based on chromatographic theory is robust and relatively simple. It is expected that the herein obtained data could be useful for selecting SPME fiber type and dimensions, estimating extraction efficiencies, as well as to develop prediction models and validate them.
... The characteristic aroma of the plant is given by the terpenes present in it. In particular, the compounds characterizing hemp are mono-and sesquiterpenes, among which β-myrcene and β-caryophyllene (Andre et al., 2016;Bertoli et al., 2010;Calvi et al., 2018;Da Porto et al., 2014;Marchini et al., 2014;Ross & Elsohly, 1996) stand out as those most present. β-myrcene is known for its antiinflammatory, analgesic and anxiolytic properties (Andre et al., 2016), while β-caryophyllene, as well as antiinflammatory, is also capable of carrying out a protective action against gastric cells. ...
Hemp seed represents an import feedstock for the future of food. Hemp is the queen plant of sustainability and brings copious yields with low demands, in terms of water, energy, and land. Moreover, is a plant renetting the soil and fits perfect in crops rotations. The food market sector of hemp seed derivatives is in a constant raise and aims to hit 5 billion of USD in the next five years. This source is so rich in nutrients and health-related compounds that just a supplementation in the final meal improves the quality of the final product. In this chapter, we present the more interesting bioactives compounds present in hemp seeds, which result abundantly in a derived food product, and could be bio-available after intestinal digestion by the host, thus generating a beneficial health-related effect.
... If the word " volatile compounds" was added together with the word " hemp" the number of documents decreased significantly to a total of 20 documents. Pellati et al., 2018); (2) industrial hemp inflorescences using supercritical CO 2 extraction and on-line fractionation (Da Porto et al., 2014); (3) oil obtained by the cold pressing of C. sativa seeds (Rovellini et al., 2013); (4) hemp seeds to check the volatile compounds profiles in order to establish markers of lipid oxidations in different types of oils (Gaca et al., 2021); (5) fresh and dried hemp flowers essential oils of C. sativa to verify how selected drying methods influence their profile (Kwásnica et al., 2020); (6) (10) fresh and dried herb, and of the resin of C. sativa in order to search for potential candidates or markers intended to trigger remote detection of hashish (Marchini et al., 2014); (11) industrial hemp residue with the aim of optimizing the extraction of cannabinoids from industrial hemp threshing residue using supercritical carbon dioxide extraction in pilot scales (Vági et al., 2019); (12) marijuana using the capillary microextractor as a sampling and preconcentration device with the objective of demonstrating that marijuana will have a distinct chemical profile, or collection of volatiles, from related plants and other products that could emit similar compounds (McPartland & Pruitt, 1999); and of (13) distillates obtained from industrial hemp hurds by thermal processing, aiming to optimize the use of hemp hurds as high-value products by utilizing slow pyrolysis for biochar and further processing for distillates (Salami et al., 2020). ...
This chapter is concerned with the sensory analysis, aroma compounds, and world regulations applied to Cannabis sativa (C. sativa). The chapter includes: (1) definitions and concepts related to sensory analysis; (2) the biosynthesis of the volatiles and the main chemical families found in this crop; (3) instrumental analysis; (4) scientific production related to C. sativa sensory quality and aroma compounds; (5) EU, USA and world regulations on its use and consumption; and (6) relevant findings in terms of sensory analysis and aroma compounds of C. sativa. Sensory analysis is the scientific discipline used to measure analyze and interpret the human reaction to products characteristics. Despite de developments already done in the instrumental analysis regarding food quality determination, the flavor sensations perceived by humans can be measured only by sensory tests. Within C. sativa, this scientific discipline can provide information about how its sensory characteristics are related to perceived quality and consumer liking, as hemp is widely used in food and cosmetics. The sensory analysis is always combined with analytical instruments to evaluate the aroma compounds. Among all these tools, for C. sativa, instrumental color, gas- and liquid- chromatography are the most used. The first one, to evaluate the color, the second one to determine terpenoids (the main chemical family of aroma compounds of hemp), and the third one is mainly used for the cannabinoids determination (compounds highly valued for their pharmacologically effect and worldwide regulated due to their psychoactive effect on humans). Finally, both sensory and instrumental analysis are required for the elucidation of C. sativa composition, as well as for their use as ingredients in food or other goods. Because instrumental analysis can help to determine the aromatic profile specific to each cultivar, while sensory analysis helps to detect which of those cultivars are more accepted in terms of flavor by the consumers. Last, both techniques might help in the optimization of hemp processing.
... Further, extraction processes such as hydrodistillation could dramatically influence the thermolabile or water-sensitive constituents. In particular, headspace sampling technique is the most suitable to describe the volatile profile of C. sativa samples [43]. Additionally, thanks to the above-described bioinformatics approach, it was possible to extrapolate important information about the investigated C. sativa inflorescences. ...
The chemical profile of the Cannabis sativa L. female inflorescences is rather complex being characterized by a large number of molecules belonging to different chemical classes. Considering the numerous applications in various fields, including the medical and pharmaceutical sectors, that have seen a large use of Cannabis genus in recent years, a precise characterization of the matrices is essential. In this regard, the application of adequate and suitable sampling and analysis techniques becomes important in order to provide an identification of the metabolites characterizing the profile of the sample under examination. The goal of this work is to provide additional information on the chemical composition of the inflorescences of five C. sativa different cultivars grown in Emilia Romagna (Italy) through the application of sophisticated analysis techniques such as Solid-Phase Microextraction-Gas Chromatography-Mass Spectrometry and Ultra-Performance Liquid Chromatography-Mass Spectrometry (SPME-GC-MS and UPLC-MS). The obtained data highlighted the presence of a high number of volatile and non-volatile compounds, thus allowing a comparative evaluation of the different samples. Furthermore, an in-depth statistical survey by Principal Components Analysis (PCA) and HeatMap, Hierarchical luster Analysis (HCA) and Partial Least Squares Discriminant Analysis (PLS-DA-VIP), was conducted to consider any correlations between the investigated cultivars. The findings of this study may help to provide more information on the C. sativa inflorescences useful for potential applications of their metabolites in scientific research.
... Moreover, the oxidation caused by UV light or heat during storage or transformation can determine the formation of oxygen-containing products as terpenoids and ketones [22,23]. In fact, secondary photo-oxidation of the terpenes can determine the generation of unstable allylic hydroperoxides whose reduction and subsequent oxidation can lead to the formation of alcohols and then aldehydes or ketones [24]. These compounds are studied for their potential toxicity, inherent to the promotion of oxidative stress [25,26]. ...
The purpose of this analytical study was to develop an advanced formulation of medical Cannabis oil (MCO) comparing the chemical profile of different extracts obtained with two existing methods (SIFAP and CALVI) and one original upgraded (CERFIT) method. Preparation methods were applied with varying solvent, temperature, and duration of the decarboxylation and extraction steps. HPLC-MS/MS TSQ and GC/FID-HS analyses were performed to investigate cannabinoid and terpene contents in the three oil extracts. Cannabinoids profile remained comparable between the formulations. CERFIT extracts exhibited a superior quantity of total terpene hydrocarbon forms (e.g., limonene and α-pinene) with no degradation occurrence (i.e., oxidized terpenes not quantifiable). Thus, this new method optimized the phytochemical profile of the MCO presenting a value opportunity to obtain a standardized high-level therapeutic product.
... Such findings have been corroborated in a more recent stud which fresh varieties expressed higher monoterpene content while dried sam demonstrated lower concentrations after a loss of the lower molecular w compounds with lower boiling points [12]. While the differences between fresh dried preparation terpenoid concentrations observed here are less prominent than i study of Ross et al. [8], this can be attributed to the advanced drying technology reg applied herein, employing low ambient temperature (vide infra, Methods Section 4. 6 The question remains whether such efforts to harness potential synergy o cannabis entourage are necessary or desirable for the cannabis consumer, irrespecti whether such use be medical or adult. Two recent studies have demonstrated a la interaction among common terpenoids on CB1 receptors [13,14] but without accoun for other mechanisms of action that might produce such synergy, whether by boo ...
Despite its use by humans for thousands of years, the technology of cannabis usage and extraction is still evolving. Given that the primary pharmacological compounds of interest are cannabinoid and terpenoids found in greatest abundance in capitate glandular trichomes of unfertilized female inflorescences, it is surprising that older techniques of hashish making have received less technological advancement. The purpose of this study was to employ organically grown cannabis and to isolate pure trichomes from freshly picked flowers via exposure to vapor from solid CO2, commonly known as “dry ice”, followed by their isolation via sifting through a 150 µ screens while maintaining the cold chain. Biochemical analysis was undertaken on fresh flower, frozen-sifted flower by-products, treated trichomes (Kryo-Kief™), dried flower, dried sifted flower by-product and dried kief. The dry ice process successfully concentrated cannabinoid content as high as 60.7%, with corresponding concentration and preservation of monoterpenoids encountered in fresh flower that are usually lost during the conventional cannabis drying and curing process. The resulting dried sifted flower by-product after dry ice processing remains a usable commodity. This approach may be of interest to pharmaceutical companies and supplement producers pursuing cannabis-based medicine development with an eye toward full synergy of ingredients harnessing the entourage effect.
... Soxhlet extraction is an old yet effective method for plant oil extraction [48]. It has been used to extract organics from over a hundred types of biomass, including Sunn hempseeds [49], marijuana cigarette [50], hashish [51], coffee [52], vanillin [53], rice bran [54], walnut kernel [55], fungal biomass [56], algal oil [57,58], orange juice [59], etc. It operates based on a continuous cycle of solvent and leaving the extracts in the extract chamber and contacting the biomass in the batch process. ...
Cannabis plant has long been execrated by law in different nations due to the psychoactive properties of only a few cannabinoids. Recent scientific advances coupled with growing public awareness of cannabinoids as a medical commodity drove legislation change and brought about a historic transition where the demand rose over ten-fold in less than five years. On the other hand, the technology required for cannabis processing and the extraction of the most valuable chemical compounds from the cannabis flower remains the bottleneck of processing technology. This paper sheds light on the downstream processing steps and principles involved in producing cannabinoids from Cannabis sativa L. (Hemp) biomass. By categorizing the extraction technology into seed and trichome, we examined and critiqued different pretreatment methods and technological options available for large-scale extraction in both categories. Solvent extraction methods being the main focus, the critical decision-making parameters in each stage, and the applicable current technologies in the field, were discussed. We further examined the factors affecting the cannabinoid transformation that changes the medical functionality of the final cannabinoid products. Based on the current trends, the extraction technologies are continuously being revised and enhanced, yet they still fail to keep up with market demands.
... (hashishene). Hashishene is formed by photo-oxidation of myrcene and was given its common name from hashish, resins of Cannabis sativa [41]. As in hashish, a similar formation of hashishene by solar radiation is conceivable during the harvest of Boswellia resins. ...
Boswellic acids, and particularly 11-keto-boswellic acids, triterpenoids derived from the genus Boswellia (Burseraceae), are known for their anti-inflammatory and potential antitumor efficacy. Although boswellic acids generally occur as α-isomers (oleanane type) and β-isomers (ursane type), 11-keto-boswellic acid (KBA) was found only as the β-isomer, β-KBA. Here, the existence and natural occurrence of the respective α-isomer, 11-keto-α-boswellic acid (α-KBA), is demonstrated for the first time. Initially, α-KBA was synthesized and characterized by high-resolution mass spectrometry (HR-MS) and nuclear magnetic resonance (NMR) spectroscopy, and a highly selective, sensitive, and accurate high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) method was developed by Design of Experiments (DoE) using a pentafluorophenyl stationary phase. This method allowed the selective quantification of individual 11-keto-boswellic acids and provided evidence for α-KBA in Boswellia spp. oleogum resins. The contents of α-KBA as well as further boswellic acids and the composition of essential oils were used to chemotaxonomically classify 41 Boswellia oleogum resins from 9 different species. Moreover, α-KBA exhibited cytotoxicity against three treatment-resistant triple-negative breast cancer (TNBC) cell lines in vitro and also induced apoptosis in MDA-MB-231 xenografts in vivo. The respective β-isomer and the acetylated form demonstrate higher cytotoxic efficacies against TNBC cells. This provides further insights into the structure-activity relationship of boswellic acids and could support future developments of potential anti-inflammatory and antitumor drugs.
... The diversity of the CsTPS genes complicates studies of gene regulation at various levels. Terpene composition can vary between cultivars, tissue types, trichome morphotypes and across development, whilst nonenzymatic modifications such as the oxidation of b-myrcene cause variation independent of genomic and transcriptomic regulation (Marchini et al., 2014;Aizpurua-Olaizola et al., 2016;Allen et al., 2019;Livingston et al., 2019;Booth et al., 2020). Consequently, future transcriptional studies would need to consider geneenvironment interactions, as well as organ, tissue and cell-type specificity. ...
Cannabis (Cannabis sativa L.) is one of the oldest cultivated plants purported to have unique medicinal properties. However, scientific research of cannabis has been restricted by the Single Convention on Narcotic Drugs of 1961, an international treaty that prohibits the production and supply of narcotic drugs except under license. Legislation governing cannabis cultivation for research, medicinal and even recreational purposes has been relaxed recently in certain jurisdictions. As a result, there is now potential to accelerate cultivar development of this multi‐use and potentially medically useful plant species by application of modern genomics technologies. Whilst genomics has been pivotal to our understanding of the basic biology and molecular mechanisms controlling key traits in several crop species, much work is needed for cannabis. In this review we provide a comprehensive summary of key cannabis genomics resources and their applications. We also discuss prospective applications of existing and emerging genomics technologies for accelerating the genetic improvement of cannabis.
... Conventional studies have led to protocols optimized for the analysis of a few targeted analytes or chemical classes. Instead, GC×GC has shown its advantages in the multiclass and high-resolution analysis of a variety of analytes [105,106]. In a recent approach, a methodology involving the use of SBSE was employed for the multiclass metabolite profiling of cannabis inflorescences [107]. ...
In this review, we consider and discuss the affinity and complementarity between a generic sample preparation technique and the comprehensive two‐dimensional gas chromatography process. From the initial technical development focus (e.g., on the GC×GC and solid‐phase microextraction techniques), the trend is inevitably shifting toward more applied challenges, and therefore, the preparation of the sample should be carefully considered in any GC×GC separation for an overreaching research. We highlight recent biomedical, food, and plant applications (2016–July 2020), and specifically those in which the combination of tailored sample preparation methods and GC×GC–MS has proven to be beneficial in the challenging aspects of non‐targeted analysis. Specifically on the sample preparation, we report on gas‐phase, solid‐phase, and liquid‐phase extractions, and derivatization procedures that have been used to extract and prepare volatile and semi‐volatile metabolites for the successive GC×GC analysis. Moreover, we also present a milestone section reporting the early works that pioneered the combination of sample preparation techniques with GC×GC for non‐targeted analysis.
... Also, terpene profiles can significantly change as a result of differential gene expression of CsTPS family during plant growth and development or in response to environmental conditions (Booth and Bohlmann, 2019;Kovalchuk et al., 2020). Although the oxygen functionality of simple terpene alcohols such as bisabolol and linalool may be caused by the enzymatic activity of CsTPS, other terpene derivatives such as caryophyllene oxide, β-elemene may result from non-enzymatic factors due to UV-or thermal-induced rearrangements or oxidation during storage or processing (Booth et al., 2017;Marchini et al., 2014). These non-enzymatic modifications can lead to more variations in terpene profiles that are independent of the cannabis genome and biochemistry. ...
Cannabis sativa L. is a high-value crop with a multi-billion dollar international market, yet due to the long history of prohibition, there is a significant lack of research on the plant and biotechnological techniques are in their infancy. Developing and applying modern techniques to Cannabis will help overcome some species-specific challenges to increase productivity and improve our knowledge about this plant. With regulatory environments relaxing in many parts of the world, there has been a significant increase in biotechnological research with this species. The current manuscript reviews the advances in Cannabis biotechnology, including molecular markers, microRNA, omics-based methods, and functional genes related to the terpene and cannabinoid biosynthesis as well as fiber quality. The foremost aim of this study is to a comprehensive review of the available literature to guide future cannabis studies in the field of genetic engineering and biotechnology.
... Most phytocannabinoids are unique to Cannabis plants, whereas terpenoids are widespread in the plant kingdom. According to the most recent literature, over 150 phytocannabinoids and 200 terpenoids have been identified in Cannabis plants (Russo, 2011;ElSohly and Gul, 2014;Marchini et al., 2014;Rice and Koziel, 2015;Hanuš et al., 2016;Berman et al., 2018;Shapira et al., 2019;Nguyen et al., 2020). ...
The therapeutic use of medical Cannabis is growing, and so is the need for standardized and therapeutically stable Cannabis products for patients. The therapeutic effects of Cannabis largely depend on the content of its pharmacologically active secondary metabolites and their interactions, mainly terpenoids and phytocannabinoids. Once harvested and during storage, these natural compounds may decarboxylate, oxidize, isomerize, react photochemically, evaporate and more. Despite its widespread and increasing use, however, data on the stability of most of the plant’s terpenoids and phytocannabinoids during storage is scarce. In this study, we therefore aimed to determine postharvest optimal storage conditions for preserving the composition of naturally biosynthesized secondary metabolites in Cannabis inflorescences and Cannabis extracts. To this end, Cannabis inflorescences (whole versus ground samples) and Cannabis extracts (dissolved in different solvents) from (-)-Δ⁹-trans-tetrahydrocannabinol- or cannabidiol-rich chemovars, were stored in the dark at various temperatures (25, 4, −30 and −80°C), and their phytocannabinoid and terpenoid profiles were analyzed over the course of 1 year. We found that in both Cannabis inflorescences and extracts, a storage temperature of 25°C led to the largest changes in the concentrations of the natural phytocannabinoids over time, making this the most unfavorable temperature compared with all others examined here. Olive oil was found to be the best vehicle for preserving the natural phytocannabinoid composition of the extracts. Terpenoid concentrations were found to decrease rapidly under all storage conditions, but temperatures lower than −20°C and grinding of the inflorescences were the least favorable conditions. Overall, our conclusions point that storage of whole inflorescences and extracts dissolved in olive oil, at 4°C, were the optimal postharvest conditions for Cannabis.
... Comprehensive 2D GC is a powerful technique used for the resolution of complex samples, with only a few reports describing its application to the static headspace of cannabis inflorescences and resins or extracted via supercritical fluids and Soxhlet. 34,35 Typical advantages of the multidimensional techniques are the superior selectivity and signal enhancement compared to a conventional separation. 36,37 In this regard, Figure S1 shows the increased resolution and selectivity provided by the 2D separation on the cannabinoids region, and Figure S2 shows the signal enhancement provided by the modulation effect on the sesquiterpene region. ...
The present research reports on the development of a methodology to unravel the complex phytochemistry of cannabis. Specifically, cannabis inflorescences were considered and stir bar sorptive extraction (SBSE) was used for the preconcentration of the metabolites. Analytes were thermally desorbed into a comprehensive two-dimensional (2D) gas chromatography (GC × GC) system coupled with low- and high-resolution mass spectrometry (MS). Particular attention was devoted to the optimization of the extraction conditions, to extend the analytes' coverage, and the chromatographic separation, to obtain a robust data set for further untargeted analysis. Monoterpenes, sesquiterpenes, hydrocarbons, cannabinoids, other terpenoids, and fatty acids were considered to optimize the extraction conditions. The response of selected ions for each chemical class, delimited in specific 2D chromatographic regions, enabled an accurate and fast evaluation of the extraction variables (i.e., time, temperature, solvent, salt addition), which were then selected to have a wide analyte selection and good reproducibility. Under optimized SBSE conditions, eight different cannabis inflorescences and a quality control sample were analyzed and processed following an untargeted and unsupervised approach. Principal component analysis on all detected metabolites revealed chemical differences among the sample types which could be associated with the plant subspecies. With the same SBSE-GC × GC-MS methodology, a quantitative targeted analysis was performed on three common cannabinoids, namely, Δ9-tetrahydrocannabinol, cannabidiol, and cannabinol. The method was validated, giving correlation factors over 0.98 and <20% reproducibility (relative standard deviation). The high-resolution MS acquisition allowed for high-confidence identification and post-targeted analysis, confirming the presence of two pesticides, a plasticizer, and a cannabidiol degradation product in some of the samples.
... β-elemene (1-methyl-1-vinyl-2,4-diisopropenyl-cyclohexane) is a derivative terpenoid found in Cannabis sativa, which may arise due to oxidation or due to thermal-or UV-induced rearrangements during processing or storage [85,178,179]. However, β-elemene is present not only in Cannabis sativa but also from Curcuma rhizome, and it is commonly used in traditional Chinese medicine due to its anticancer properties with no reported severe side effects [180]. ...
Medicinal use of Cannabis sativa L. has an extensive history and it was essential in the discovery of phytocannabinoids, including the Cannabis major psychoactive compound—Δ9-tetrahydrocannabinol (Δ9-THC)—as well as the G-protein-coupled cannabinoid receptors (CBR), named cannabinoid receptor type-1 (CB1R) and cannabinoid receptor type-2 (CB2R), both part of the now known endocannabinoid system (ECS). Cannabinoids is a vast term that defines several compounds that have been characterized in three categories: (i) endogenous, (ii) synthetic, and (iii) phytocannabinoids, and are able to modulate the CBR and ECS. Particularly, phytocannabinoids are natural terpenoids or phenolic compounds derived from Cannabis sativa. However, these terpenoids and phenolic compounds can also be derived from other plants (non-cannabinoids) and still induce cannabinoid-like properties. Cannabimimetic ligands, beyond the Cannabis plant, can act as CBR agonists or antagonists, or ECS enzyme inhibitors, besides being able of playing a role in immune-mediated inflammatory and infectious diseases, neuroinflammatory, neurological, and neurodegenerative diseases, as well as in cancer, and autoimmunity by itself. In this review, we summarize and critically highlight past, present, and future progress on the understanding of the role of cannabinoid-like molecules, mainly terpenes, as prospective therapeutics for different pathological conditions.
... For example, P1 samples were particularly abundant in β-farnesene, while in plant 3 (both P and M samples) it was absent. All samples contained longicyclene, the sesquiterpene rarely identified in C. sativa inflorescences, with the exception of one drug-type chemovar [31]. ...
The phytochemical profiling of hemp inflorescences of clonal plants growing in different conditions related to altitude was investigated. Four strains of industrial hemp (Cannabis sativa L., family Cannabaceae) of Kompolti variety were selected and cloned to provide genetically uniform material for analyses of secondary metabolites (terpenes, cannabinoids, and flavonoids) at two different elevations: mountain (Alagna Valsesia 1200 m ASL) and plains (Vercelli Province 130 m ASL). Environmental conditions influenced by elevation have proven to be important factors inducing variations in hemp inflorescences’ secondary metabolite composition. In fact, all plants grown at altitude exhibited a higher total amount of terpenes when compared with plains counterparts, with β-Myrcene, trans-Caryophyllene and α-Humulene as the main contributors. A metabolomic, un-targeted approach performed by HPLC-Q-Exactive-Orbitrap®-MS platform with subsequent data processing performed by Compound Discoverer™ software, was crucial for the appropriate recognition of many metabolites, clearly distinguishing mountain from plains specimens. Cannabidiolic acid CBDA was the most abundant phytocannabinoid, with significantly higher concentrations in the mountain samples. The metabolic pathway of CBGA (considered as the progenitor/precursor of all cannabinoids) was also activated towards the production of CBCA, which occurs in considerably 3 times higher quantities than in the clones grown at high altitude. Isoprenoid flavones (Cannaflavins A, B, and C) were correspondingly upregulated in mountain samples, while apigenin turned out to be more abundant in plains samples. The possibility to use hemp inflorescences in pharmaceutical/nutraceutical applications opens new challenges to understand how hemp crops respond in terms of secondary metabolite production in various environments. In this regard, our results with the applied analytical strategy may constitute an effective way of phytochemical profiling hemp inflorescences.
... (Rice and Koziel, 2015a, b). The principle (trace) components are reported to be αand β-pinene, β-myrcene, dlimonene, cis-ocimene, β-caryophyllene, β-farnesene, and α-humulene (Hood et al., 1973;Turner et al., 1980;Hillig, 2004;Fischedick et al., 2010;Martyny et al., 2013;Marchini et al., 2014;Rice and Koziel, 2015a). The precise mix of chemical species, however, was strongly dependent on strain and the growing conditions (Fischedick et al., 2010). ...
The legal commercialization of cannabis for recreational and medical use has effectively created a new and almost unregulated cultivation industry. In 2018, within the Denver County limits, there were more than 600 registered cannabis cultivation facilities (CCFs) for recreational and medical use, mostly housed in commercial warehouses. Measurements have found concentrations of highly reactive terpenes from the headspace above cannabis plants that, when released in the atmosphere, could impact air quality. Here we developed the first emission inventory for cannabis emissions of terpenes. The range of possible emissions from these facilities was 66–657 t yr-1 of terpenes across the state of Colorado; half of the emissions are from Denver County. Our estimates are based on the best available information and highlight the critical data gaps needed to reduce uncertainties. These realizations of inventories were then used with a regulatory air quality model, developed by the state of Colorado to predict regional ozone impacts. It was found that most of the predicted changes occur in the vicinity of CCFs concentrated in Denver. An increase of 362 t yr-1 in terpene emissions in Denver County resulted in increases of up to 0.34 ppb in hourly ozone concentrations during the morning and 0.67 ppb at night. Model predictions indicate that in Denver County every 1000 t yr-1 increase in terpenes results in 1 ppb increase in daytime hourly ozone concentrations and a maximum daily 8 h average (MDA8) increase of 0.3 ppb. The emission inventories developed here are highly uncertain, but highlight the need for more detailed cannabis and CCF data to fully understand the possible impacts of this new industry on regional air quality.
... Considering the terpenoids fraction, our results are qualitatively comparable with those reported by others (Bertoli et al., 2010;Elzinga et al., 2015;Aizpurua-Olaizola et al., 2016;Benelli et al., 2018, Namdar et al., 2018. The particularity of both chemovars presented in this study is the presence of the strong signal of sesqui-CBG accompanied with very reach sesquiterpenes fraction, indicating that geographic origin accompanied with environmental conditions is one of the important variables that determine the terpene fingerprint (Giorgi et al., 2013b;Marchini et al., 2014;Giupponi et al., 2017). The presumed appearance of fenchyl-cannabidiolate might also be defined as possible discrimination factor that concerns cultivation/ecological conditions. ...
Hemp (Cannabis sativa L.) is a multifunctional crop that is capable of prompt environmental adaptation. In this study, a monoecious cultivar (Futura 75) and a dioecious one (Finola) were tested in a mountain area in Valsaviore (Rhaetian Alps, Italy; elevation: 1,100 m a.s.l.) during the growing season 2018. Phytochemical behavior was evaluated by different analytical approaches: HPLC-high-resolution mass spectrometry, SDS-PAGE LC-MS/MS, HS-SPME GC-MS, and GC-FID in order to obtain complete profile of two varieties cultivated in altitude. CSR functional strategy used for ecological evaluation revealed that both genotypes are mainly competitors, although Finola is more stress tolerator (C:S:R = 57:26:17%) than Futura (C:S:R = 69:15:16%). The Finola inflorescences were characterized by higher quantities of β-ocimene and α-terpinolene, while α- and ß-pinene accompanied by extremely high ß-myrcene were found as predominant in Futura. Both varieties were rich in sesquiterpenes (45 recognized) among which trans-caryophyllene and α-humulene were the most abundant. Total tetrahydrocannabinol level was lower than 0.1%, while the most abundant cannabinoid was cannabidiolic acid (CBDA): 2.3% found in Finola vs. 2.7% revealed for Futura. The level of corresponding neutral form, cannabidiol, varied drastically: 0.27% (Finola) vs. 0.056% (Futura). Finola showed the unique cannabinoid profile with unexpectedly high cannabidivarin, 2-fold higher that corresponding acidic analogue, whereas the particularity of Futura 75 was the occurrence of cannabigerolic acid (CBGA) in the quantities that was double than those exposed for Finola. The seeds from both chemovars proved to be rich in polyunsaturated fatty acids, and Finola showed a higher ratio ω6/ω3. No difference was found in the protein content, and the SDS-PAGE profile was similar. The most abundant protein was edestin, followed by heat shock protein 70, ß-conglycinin, and vicilin. In conclusion, comprehensive phytochemical and ecological study of two fiber-type varieties cultivated in Italian Alps displayed specific, legal, and safe cannabinoids profile, followed by particular terpene composition, polyunsaturated fatty acids content, and favorable protein profile. This postulates that geographical provenience of hemp should be considered in selecting a variety that would be suitable for a specific end-use nutraceutical application.
... GC × GC provides about a 10-fold increase in peak capacity versus one-dimensional GC, and significantly more peaks are produced in a given separation [4]. Hence, GC × GC is well suited for separating complex samples containing hundreds of analytes, and has been adapted for applications in diverse fields such as forensics [5,6], food chemistry [7], metabolomics [8,9], wastewater testing [10], and analysis of diesel fuels and biofuels [11][12][13]. However, for complex sample separations, it becomes difficult to visually ascertain differences between different samples, such as in the comparative analysis of sample classes. ...
Principal component analysis (PCA) is a widely applied chemometric tool for classifying samples using comprehensive two-dimensional (2D) gas chromatography (GC × GC) separation data. Classification via PCA can be improved by 2D binning of the data. A "standard operating procedure (SOP) bin size" is often applied to improve the S/N and to mitigate potential retention time misalignment issues. The SOP bin size is generally selected to be slightly larger than the typical 2D peak dimensions. In this study we examine to what extent a single SOP bin size is optimal for all of the class comparisons that can be made in a single PCA scores plot. For this purpose, a GC × GC-FID dataset comprised of 5 different diesel fuels (i.e., 5 sample classes), each run with 4 replicates using a reverse column configuration (polar 1D column and non-polar 2D column) was utilized. The dataset was collected within about one day, which minimized retention time misalignment in order to allow the study to focus on S/N enhancement concurrent with maintaining the chemical selectivity provided by the GC × GC separations. A total of 110 bin sizes were evaluated. Degree-of-class separation (DCS) was utilized as a quantitative metric to assess the impact of binning in improving separation in the scores plot. The DCS was calculated pair-wise between nearest neighbor sample classes for each of the 5 sample classes in the scores plot (5 sample class pairs). Results indicated the SOP bin size did not provide the highest DCS for any of the 5 fuel pairs. Each fuel pair is found to have its own optimal bin size, suggesting the binning finds the balance between S/N optimization concurrent with leveraging the chemical selectivity information differences in the samples as manifested in their GC × GC separation "patterns". Robustness of the findings in this study were supported by leaving out one fuel at a time and re-running the PCA models.
... Sabinene is a natural liquid monoterpene obtained from essential oils of various plants, including Cannabis. The chemical formula of sabinene is C 10 H 6 and it has a bicyclic structure [9]. Several studies have reported the biological functions of sabinene, including its antifungal [10] and anti-inflammatory properties [11]. ...
Background: Streptococcus mutans is one of the most important cariogenic bacteria associated with dental caries. Sabinene is a major component of several herbal essential oils. However, the anti-cariogenic effects of sabinene and the underlying mechanism remain to be elucidated.
Objectives: We investigated the inhibitory effects of sabinene on the cariogenic activity and studied the underlying mechanism.
Design: S. mutans were treated with various concentrations of sabinene and the inhibitory effects were evaluated based on the bacterial growth, acid production and biofilm formation. Real-time polymerase chain reaction (PCR) was performed for several virulence factors.
Results: The growth and adherence of S. mutans were inhibited by sabinene. Consistent with the inhibitory effects on bacterial adhesion, gbpB level significantly decreased. Acid production and biofilm formation was also inhibited. In line with the inhibitory effects of sabinene on biofilm formation and pH tolerance, real-time PCR results showed the down regulation in the expression levels of gtfB, gtfC, gtfD, vicR, brpA, and relA. Moreover, high concentrations of sabinene exhibited bactericidal activity.
Conclusion: Together our results suggest that sabinene serves as a useful component in the inhibition of the cariogenic activity of S. mutans, indicative of its possible applications in the development of oral healthcare products.
... The GC data of the hemp EOs analysed in this work are in accordance with those previously described in the literature [5,[14][15][16][17][18][19][20][21][25][26][27][28][29]. Table 1. ...
Volatile terpenes represent the largest group of Cannabis sativa L. components and they are responsible for its aromatic properties. Even if many studies on C. sativa have been focused on cannabinoids, which are terpenophenolics, little research has been carried out on its volatile terpenic compounds. In the light of all the above, the present work was aimed at the chemical characterization of seventeen essential oils from different fibre-type varieties of C. sativa (industrial hemp or hemp) by means of GC-MS and GC-FID techniques. In total, 71 compounds were identified, and the semi-quantitative analysis revealed that α- and β-pinene, β-myrcene and β-caryophyllene are the major components in all the essential oils analysed. In addition, a GC-MS method was developed here for the first time, and it was applied to quantify cannabinoids in the essential oils. The antibacterial activity of hemp essential oils against some pathogenic and spoilage microorganisms isolated from food and food processing environment was also determined. The inhibitory effects of the essential oils were evaluated by both the agar well diffusion assay and the minimum inhibitory concentration (MIC) evaluation. By using the agar diffusion method and considering the zone of inhibition, it was possible to preliminarily verify the inhibitory activity on most of the examined strains. The results showed a good antibacterial activity of six hemp essential oils against the Gram-positive bacteria, thus suggesting that hemp essential oil can inhibit or reduce bacterial proliferation and it can be a valid support to reduce microorganism contamination, especially in the food processing field.
... Other terpene derivatives detected in cannabis may arise non-enzymatically due to oxidation or due to thermal-or UV-induced rearrangements during processing or storage, such as caryophyllene oxide, βelemene, or derivatives of myrcene [8,12]. These non-enzymatic modifications may add a level of variation that is independent of the plant genome and biochemistry. ...
Cannabis sativa (cannabis) produces a resin that is valued for its psychoactive and medicinal properties. Despite being the foundation of a multi-billion dollar global industry, scientific knowledge and research on cannabis is lagging behind compared to other high-value crops. This is largely due to legal restrictions that have prevented many researchers from studying cannabis, its products, and their effects in humans. Cannabis resin contains hundreds of different terpene and cannabinoid metabolites. Many of these metabolites have not been conclusively identified. Our understanding of the genomic and biosynthetic systems of these metabolites in cannabis, and the factors that affect their variability, is rudimentary. As a consequence, there is concern about lack of consistency with regard to the terpene and cannabinoid composition of different cannabis ‘strains’. Likewise, claims of some of the medicinal properties attributed to cannabis metabolites would benefit from thorough scientific validation.
... A standardised protocol for oily preparations is therefore also required, but until now has not been formulated. In this context, cannabis extraction was performed using olive oil and a standardised medicinal cannabis "flos" (according to pharmaceutical standards) [31,34,39,44,45]. ...
... GC-FID and GC/MS are the most frequently used methods for the analysis of volatile terpenes [26][27][28][29][30]. Other gas chromatography techniques such as HS GC/FID, HS GC/MS, two-dimensional (GC × GC/qMS), HS solid-phase microextraction (HS-SPME), GC/ MS, and GC × GC/MS were also used for the analysis of cannabis and hashish samples [31][32][33]. Romano and Hazekamp compared five different methods for cannabis oil preparation (using naphtha, petroleum ether, ethanol, and two types of olive oil). They performed the analysis of both terpenes and cannabinoids using GC-FID and HPLC-UV, respectively [34]. ...
Terpenes are the major components of the essential oils present in various Cannabis sativa L. varieties. These compounds are responsible for the distinctive aromas and flavors. Besides the quantification of the cannabinoids, determination of the terpenes in C. sativa strains could be of importance for the plant selection process. At the University of Mississippi, a GC-MS method has been developed and validated for the quantification of terpenes in cannabis plant material, viz., α-pinene, β-pinene, β-myrcene, limonene, terpinolene, linalool, α-terpineol, β-caryophyllene, α-humulene, and caryophyllene oxide. The method was optimized and fully validated according to AOAC (Association of Official Analytical Chemists) guidelines against reference standards of selected terpenes. Samples were prepared by extraction of the plant material with ethyl acetate containing n-tridecane solution (100 µg/mL) as the internal standard. The concentration-response relationship for all analyzed terpenes using the developed method was linear with r2 values > 0.99. The average recoveries for all terpenes in spiked indoor cultivated samples were between 95.0 – 105.7%, with the exception of terpinolene (67 – 70%). The measured repeatability and intermediate precisions (% relative standard deviation) in all varieties ranged from 0.32 to 8.47%. The limit of detection and limit of quantitation for all targeted terpenes were determined to be 0.25 and 0.75 µg/mL, respectively. The proposed method is highly selective, reliable, and accurate and has been applied to the simultaneous determination of these major terpenes in the C. sativa biomass produced by our facility at the University of Mississippi as well as in confiscated marijuana samples.
... As of now, the majority of the published studies dealt with the evaluation of the cannabinoid contents of seized hashish samples [6,7], but a wider analysis of the chemical profile of the samples might be even more interesting to assess the chemical links between different seizures. The evaluation of a larger set of compounds as a tool to discriminate between different sources has been used for the raw plant material; the VOC emission profiles of different Cannabis sativa L. samples have been analyzed, proving that it is a suitable method for the comparison of samples of cannabis seized in different places [8,9]. The present study addresses the need of a fast, solventless, easy and reliable analysis method, namely the VOC analysis, for a cannabis manufactured form; the VOC profiles of 48 hashish samples confiscated by Guardia di Finanza (an Italian law enforcement agency) have been analyzed by means of headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). ...
Purpose
The statistical evaluation of the chemical profile of seized hashish samples is a valuable tool to aid the estimation of the route through which the material has reached the dealers’ market.
Methods
In this study, the complete volatile organic compound (VOC) emission profiles of 48 seized hashish samples have been analyzed by means of headspace solid-phase microextraction/gas chromatography–mass spectrometry and evaluated with chemometric tools; multivariate statistical analyses, both hierarchical cluster analysis and principal component analysis (PCA) methods have been performed on the results to assess the existence of possible patterns throughout the samples.
Results
The total VOC emission profiles sharply distributed the samples in clusters based on their batches of origin; this trend was also clearly shown in the PCA plot, in which samples coming from the same seizure were grouped together. The Δ⁹-tetrahydrocannabinol (THC) content analysis did not show a relevant trend in terms of lot of origin of the samples.
Conclusions
The evaluation of the VOCs released into the headspace traced a much more complete chemical profiling of the samples, as compared to the analysis of cannabinoids only, or the THC titration. The multivariate statistical analyses were very useful to estimate the origin of the seized material.
Water availability is a key factor affecting both morphological development and secondary metabolite production in Cannabis sativa L. This study evaluated the effects of water stress applied during the vegetative and flowering stages on plant performance, cannabinoid concentration, and terpene composition in two Chemotype III (cannabidiol-dominant) varieties. Plants were subjected to moderate and severe water stress, and responses were assessed through biomass measurements, GC-MS analyses, and multivariate statistics. Water stress significantly influenced biomass allocation, with increased dry biomass but reduced harvest index, particularly under flowering-stage stress. Cannabidiol (CBD) content declined with increasing stress, while tetrahydrocannabinol (THC) levels increased under vegetative stress, indicating a stress-induced shift in cannabinoid biosynthesis. Cannabinol (CBN) levels also increased, suggesting enhanced THC degradation. Terpene composition was predominantly genotype-driven. PCA-MANOVA showed significant effects of variety, stress level, and their interaction, yet only minor volatiles were modulated by stress, while the most abundant terpenes remained stable across treatments, preserving the varietal aroma profile. These results underline the importance of genetic background and irrigation timing in determining cannabis yield and quality. Optimized water management is essential to ensure phytochemical consistency and sustainable production, especially in high-value medicinal and aromatic applications.
In 2019, an air emissions field sampling study was conducted by the Colorado Department of Public Health and Environment's Air Pollution Control Division (APCD) at four commercial cannabis cultivation facilities. Measurements of ambient biogenic volatile organic compounds (VOC) concentrations were collected from various growing stages of cannabis (vegetative and flowering) and during post-harvest activities (drying and trimming). This data was then used to determine room-specific biogenic VOC emission rates for three of the facilities from the vegetative stage of the life cycle through post-harvest activities. This study shows that the magnitude of biogenic VOC emissions within a cannabis cultivation facility varies widely with the highest emission rates of up to 7.18E-1 kg/hr found during mechanical trimming and up to 2.33E-1 kg/hr in the drying rooms. These were up to an order of magnitude higher than emission rates found in the cultivation rooms. For example, Facility A vegetative room had an emissions rate of 1.46E-2 kg/hr. Normalized by the amount of biomass present, the drying rooms had the highest VOC emissions rates, with a maximum rate of 1.6E-3 kg/hr/kg biomass. The flowering room rates were found to be up to 3.25E-4 kg/hr/kg biomass and drying rooms up to 1.16E-3 kg/hr/kg biomass. When normalized by plant count, emission rates in the flower rooms ranged from 8.11E-6 to 3.62E-4 kg/hr/plant. The dominant monoterpenes from sampling were β-myrcene, terpinolene, and D-limonene. These data suggest that the variability in emission rates across cannabis production will create a challenge in establishing a generalized emission factor for all facilities. Across the industry, cannabis cultivation conditions and strategies can vary widely impacting the amount and type of VOC emissions. Minimizing uncertainties for VOC emission from cannabis facilities requires site specific information on air exchange rates, plant counts, cannabis strains, biomass, and if hand or mechanical processing is used.Implications This study found that the magnitude of biogenic VOC emissions within a cannabis cultivation varies widely throughout rooms found in the facility, with the highest emissions found during post-harvest activities (i.e. trimming) and the lowest rates in the vegetative room. These data suggest that the large emission sources of VOCs are found post-harvest and emission inventories based solely on cultivation emissions will underestimate total biogenic VOC emissions from indoor cannabis cultivation facilities. The dominant measured terpenes throughout all facilities from cultivation to post harvest were: β-myrcene, terpinolene, and D-limonene.
Absolute content of terpenes in inflorescences of two strains of Cannabis sativa L., CAT 1 and CAT 3, has been determined. Twenty terpenes commonly present in these samples were quantified by solid phase microextraction combined with gas chromatography and flame ionization detection (SMPE/GC-FID). High amounts of β-myrcene, α-pinene, β-pinene, limonene, (E)-β-ocimene, β-caryophyllene, α-humulene, (E)-nerolidol, and linalool, were found in both strains. Lower concentrations (< 20 µg·g⁻¹) of other terpenes were also determined. Only (E)-β-ocimene was detected at 50 µg·g⁻¹ in CAT 3 whereas it was below the LOD in CAT 1. Concentrations of other compounds for which standards were not available, were estimated based on a response factor obtained from the calibration curves of compounds with similar chemical structures. Fingerprints of both CAT strains were obtained and the identities of most volatile compounds were assigned using gas chromatography coupled to mass spectrometer detector (GC-MS).
Additionally, an assessment of variability of terpenes was achieved by analyzing ten plants of each strain grown under controlled conditions and harvested at the same time. This variability was about 20 %, considering terpenes at concentration above 20 µg·g ⁻¹.
In 2019, an air emissions field sampling study was conducted by the Colorado Department of Public Health and Environment (CDPHE) Air Pollution Control Division (APCD) at three commercial cannabis cultivation facilities. The goal of the study was to quantify biogenic-terpene volatile organic compound (VOC) emissions from growing cannabis at cultivation facility exhaust points to estimate a VOC emission rate by a top-down approach. The resulting VOC emission rates were then used in combination with 2019 commercial cannabis cultivation facility biomass production volumes (harvest weight) and cultivation locations from the Colorado Department of Revenue's Marijuana Enforcement Division (MED) to model the potential ozone and PM2.5 formation impacts of the cannabis industry in the Denver Metro North Front Range (DM/NFR) Ozone Nonattainment Area (NAA).Despite cannabis cultivation facilities' high nuisance odors, this study found the biogenic VOC emissions rate from the sampled indoor facilities to be low (2.13 lbs to 11.12 lbs of VOC/ton of cannabis harvested), even at large production facilities. The dominant terpenes from this sampling study present in most samples were β-caryophyllene, D-limonene, terpinolene, α-pinene, β-pinene and β-myrcene respectively by concentration. Interestingly, the cannabis emissions exhaust profile lacked isoprene, a terpene commonly emitted from other plants that is highly reactive and has great potential to contribute to ozone formation [Sharkey et al 2007, Wang et al. 2019]. The low biogenic VOC emissions rate and the lack of isoprene from the cannabis cultivation facilities sampled resulted in a very low to negligible impact on both ozone formation (0.005% - 0.009% increase in ozone from cannabis cultivation) and PM2.5 formation (largest maximum 24-hour PM2.5 difference of 0.009 µg/m3) in the DM/NFR NAA.
Marijuana, or Cannabis sativa L., is a common psychoactive plant used for both recreational and medicinal purposes. In many countries, cannabis-based medicines have been legalized under certain conditions because of their immense prospects in medicinal applications. With a comprehensive insight into the prospects and challenges associated with the pharmacological use and global trade of C. sativa, this mini-review focuses on the medicinal importance of the plant and its legal status worldwide; the pharmacological compounds and its therapeutic potential along with the underlying public health concerns and future perspective are herein discussed. The existence of major compounds including Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol, cannabinol, and cannabichromene contributes to the medicinal effects of the cannabis plant. These compounds are also involved in the treatment of various types of cancer, epilepsy, and Parkinson's disease displaying several mechanisms of action. Cannabis sativa is a plant with significant pharmacological potential. However, several aspects of the plant need an in-depth understanding of the drug mechanism and its interaction with other drugs. Only after addressing these health concerns, legalization of cannabis could be utilized to its full potential as a future medicine.
The global Cannabis Sativa market, including essential oils, foods, personal-care products, and medical formulations has gained much attention over the last years due to the favorable regulatory framework. Undoubtedly, the enormous interest about cannabis cultivation mainly derives from the well-known pharmacological properties of cannabinoids and terpenes biosynthesized by the plants. In this review, the most recently used analytical methodologies for detecting both cannabinoids and terpenes are described. Well-established and innovative extraction protocols, and chromatographic separations, such as GC and HPLC, are reviewed highlighting their respective advantages and drawbacks. Lastly, GC × GC techniques are also reported for accurate identification and quantification of terpenes in complex cannabis matrices.
In this work is proposed a critical review of the scientific literature about the extraction of products of industrial interest from Cannabis sativa L., like cannabinoids, essential oil and seed oil, using supercritical and subcritical CO2. Traditional techniques are also reviewed and critically discussed to evidence the advantages of CO2 processing. The extracts can be also used in pharmaceutical and biomedical formulations, in form of co-precipitates and capsules, to improve active compounds bioavailability and performance. Further studies can be required, mainly based on the analysis of mass transfer resistances during extraction and on solubility data of the compounds to be extracted, to improve process selectivity and the purity of the extracts obtained.
Background: Marijuana blunts, which are tobacco cigar wrappers filled with marijuana, are commonly smoked in the US as a means of cannabis use. The use of marijuana blunts presents toxicity concerns because the smoke contains both marijuana-related and tobacco-related chemicals. Thus, it is important to understand the chemical composition of mainstream smoke (MSS) from marijuana blunts. This study demonstrates the ability to detect and identify chemical constituents exclusively associated with blunt MSS in contrast to tobacco cigar MSS (designated as ‘new exposures’) through non-targeted chemical analysis.
Methods: Samples collected separately from blunt MSS and tobacco cigar MSS were analyzed using two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC-TOFMS).
Results and Discussion: Two new exposures, which likely represent only a subset of all new exposures, were identified by evaluating the data from thousands of detected signals and then confirming selected compound identities in analyses using authentic chemical standards. The two confirmed new exposures, mellein and 2-phenyl-2-oxazoline, are not cannabinoids and, to the best of our knowledge, have not been previously reported in association with cannabis, tobacco, or smoke of any kind. In addition, we detected and quantified three phenols (2-, 3-, and 4-ethylphenol) in blunt MSS. Given the toxicity of phenols, quantifying the levels of other phenols could be pursued in future research on blunt MSS.
Conclusion: This study shows the power and utility of GC × GC-TOFMS as a methodology for non-targeted chemical analysis to identify new chemical exposures in blunt MSS and to provide data to guide further investigations of blunt MSS.
The oil of the seeds, petroleum ether and methanol extracts of the whole plant of Cannabis sativa belonging to the family Cannabinaceae were screened for their antimicrobial activity against two Gram positive organisms (Bacillus subtilis, Staphylococcus aureus), two Gram negative organisms (Escherichia coli, Pseudomonas aeruginosa) and two fungi namely Aspergillus niger and Candida albicans using the cup plate agar diffusion method. The oil of the seeds of Cannabis sativa exerted pronounced antibacterial activity (21-28 mm) against Bacillus subtilis and Staphylococcus aureus, moderate activity (15 mm) against Escherichia coli and high activity (16 mm) against Pseudomonas aeruginosa and inactive against the two fungi tested. The petroleum ether extract of the whole plant exhibited pronounced antibacterial activity (23-28 mm) against both Bacillus subtilis and Staphylococcus aureus organisms, high activity (16 mm) against Escherichia coli and inactive against Pseudomonas aeruginosa and both fungi. The methanol extract of the whole plant showed also pronounced antibacterial activity (29 mm) against Bacillus subtilis, low activity (12 mm) against Staphylococcus aureus and high activity (16-18 mm) against both Gram negative organisms, inactive against Aspergillus niger and low activity (13 mm) against Candida albicans. The minimum inhibitory concentrations of Cannabis sativa methanol extracts of the seeds and the whole plant against the standard organisms were determined using the agar plate dilution method. The standard organisms were tested against reference antibacterial and antifungal drugs and the results were compared with the activity of the extracts.
We review the literature on comprehensive two-dimensional gas chromatography (GC × GC), emphasizing developments in the period 2003–2005. The review opens with a general introduction, the principles of the technique and the set-up of GC × GC systems. It also discusses theoretical aspects, trends in instrumentation, column combinations, and detection techniques – notably mass spectrometric detection. We devote attention to a wide variety of applications and to analytical performance.Part II discusses the key aspects of modulation and the various modes of detection.
We report on a research aimed at evaluating the capacity of a simple, low-cost, portable electronic nose system based on commercially available metal oxide gas sensors to classify different types of drugs. Five drugs, namely cannabis buds, cannabis plants, hashish, snuff tobacco and tobacco leaves were employed. A dedicated real-time data acquisition system based on dynamic headspace sampling, a microcontroller and a laptop computer have been designed and constructed for this application. To demonstrate its discrimination capability, unsupervised and supervised classification models have been built and validated. Principal Component Analysis (PCA) of volatile profiles revealed five distinct groups corresponding to the five different drugs analyzed. This was further confirmed by a multivariate analysis of variance (MANOVA) test. Support Vectors Machines (SVMs) were applied to build a classifier and reached a 98.5% success rate in the recognition of the different drugs analyzed. This work demonstrates for the first time that the electronic nose technology could be successfully applied to the identification of illegal drugs.
We review the literature on comprehensive two-dimensional gas chromatography (GC × GC), emphasizing developments in the period 2003–2005. The review opens with a general introduction, the principles of the technique and the set-up of GC × GC systems. It also discusses theoretical aspects, trends in instrumentation, column combinations, and detection techniques – notably mass spectrometric detection. We devote attention to a wide variety of applications and to analytical performance.
We review the literature on comprehensive two-dimensional gas chromatography (CC x GC), emphasizing developments in the period 2003-2005. The review opens with a general introduction, the principles of the technique and the set-up of CC x CC systems. It also discusses theoretical aspects, trends in instrumentation, column combinations, and detection techniques notably mass spectrometric detection. We devote attention to a wide variety of applications and to analytical performance. (c) 2006 Elsevier Ltd. All rights reserved.
We review the literature on comprehensive two-dimensional gas chromatography (GC x GC), emphasizing developments in the period 2003-2005. The review opens with a general introduction the principles of the technique and the set-up of GC x GC systems. It also discusses theoretical aspects, trends in instrumentation, column combinations, and detection techniques notably mass spectrometric detection. We devote attention to a wide variety of applications and to analytical performance. (c) 2006 Elsevier Ltd. All rights reserved.
Current commercially available mimics contain varying amounts of either the actual explosive/drug or the chemical compound of suspected interest by biological detectors. As a result, there is significant interest in determining the dominant chemical odor signatures of the mimics, often referred to as pseudos, particularly when compared to the genuine contraband material. This dissertation discusses results obtained from the analysis of drug and explosive headspace related to the odor profiles as recognized by trained detection canines. Analysis was performed through the use of headspace solid phase microextraction in conjunction with gas chromatography mass spectrometry (HS-SPME-GC-MS). Upon determination of specific odors, field trials were held using a combination of the target odors with COMPS. Piperonal was shown to be a dominant odor compound in the headspace of some ecstasy samples and a recognizable odor mimic by trained detection canines. It was also shown that detection canines could be imprinted on piperonal COMPS and correctly identify ecstasy samples at a threshold level of approximately 100ng/s. Isosafrole and/or MDP-2-POH show potential as training aid mimics for non-piperonal based MDMA. Acetic acid was shown to be dominant in the headspace of heroin samples and verified as a dominant odor in commercial vinegar samples; however, no common, secondary compound was detected in the headspace of either. Because of the similarities detected within respective explosive classes, several compounds were chosen for explosive mimics. A single based smokeless powder with a detectable level of 2,4-dinitrotoluene, a double based smokeless powder with a detectable level of nitroglycerine, 2-ethyl-1-hexanol, DMNB, ethyl centralite and diphenylamine were shown to be accurate mimics for TNT-based explosives, NG-based explosives, plastic explosives, tagged explosives, and smokeless powders, respectively. The combination of these six odors represents a comprehensive explosive odor kit with positive results for imprint on detection canines. As a proof of concept, the chemical compound PFTBA showed promise as a possible universal, non-target odor compound for comparison and calibration of detection canines and instrumentation. In a comparison study of shape versus vibration odor theory, the detection of d-methyl benzoate and methyl benzoate was explored using canine detectors. While results did not overwhelmingly substantiate either theory, shape odor theory provides a better explanation of the canine and human subject responses.
A sample of hashish was extracted consecutively with petroleum ether, benzene, and methanol. When tested intravenously in
monkeys only the petroleum-ether fraction was active. This material was further fractionated. The only active compound isolated
was Δ1-tetrahydrocannabinol. Cannabinol, cannabidiol, cannabichromene, cannabigerol, and cannabicyclol when administered together
with Δ1-tetrahydrocannabinol do not cause a change in the activity of the latter, under the experimental conditions used. These results
provide evidence that, except for Δ1-tetrahydrocannabinol, no other major, psychotomimetically active compounds are present in hashish.
A comparison of real versus simulated contraband volatile organic compound (VOCs) for reliable detector dog training utilizing (solid-phase microextraction) SPME-GC-MSwas done with the number of field studies. Detector dog teams are trained to detect most commonly used narcotics as cocaine, and chemical explosives. The GC-MS analysis was performed on an Agilent 6890 series with HP 5973 mass spectrometer, where the samples were placed in a line format. The study uses the data sheet as the positive control and returned a 100% alert response from the canines, exhibiting the dissipation rate for each of the compounds tested. The results of the tests were found with commercially available products as the Sigma Pseudoscents and NESTT, and showed that these do not create the same volatile odor compounds as the contraband parent compounds. The overall results present improvement in the performance and standardization of biological and instrumental standoff detection of targets.
The detection and identification of volatile compounds is essential to the successful undertaking of numerous forensic analyses. Biological olfactory systems possess the extraordinary ability to not only detect many thousands of distinct volatile compounds (odors) but also to discriminate between them. Whole-organism biological sensors, such as detection canines, have been employed in forensic science as volatile compound detectors for many years. A variety of insects including bees, wasps, and moths, which have also been shown to detect volatile compounds of forensic significance, have been investigated for their potential application in field-based detection systems. While the fundamental aim for many developers of portable instruments is to replicate the remarkable ability of biological olfactory systems, such analytical equipment is yet to possess the detection and discriminatory powers achieved by biological sensors. Recent literature reveals an increasing interest in olfactory receptors - the biological components that impart olfactory ability - for detecting volatile compounds associated with forensically significant substances such as explosives and illicit drugs. This paper reviews the literature regarding the current, and potential future, use of biological organisms as sensors for forensic science applications.
The separation and identification of some new substances in the essential oil of cannabis sativa [Longifolen (I), Humulenepoxid-I (II) and - II (III), Caryophyllenol-I (IV), m-Mentha-1.8(9)-dien-5-ol (V)] are described.
Both cationic and anionic clays, with sensitizers anchored onto their interlayer by ionic linkage, are found to be useful ‘microreactors’ to achieve triplet state reactivity of myrcene in an experimentally simple, inexpensive and clean reaction. These clay-bound sensitizers are characterized by X-ray and their advantages are highlighted.
The volatile composition of ten fibre hemp (Cannabis sativa L.) varieties was investigated during two successive growing seasons under temperate climatic conditions in Central Italy.The freshly plant inflorescences were hydrodistilled and the essential oils (EOs) were characterized by GC–MS. In addition, the composition of the aroma emitted spontaneously from the freshly plant inflorescences were analysed by SPME-GC–MS. The EO yields of eight dioecious (Carmagnola, C.S., Red Petiole, Pop 1, Pop 2, Pop 3, Pop 4, Pop 5) and two monoecious (Codimono and Felina 34) cultivars ranged from 0.11 to 0.25% (w/w) and showed a significant production of α-pinene (3–20%), β-pinene (1–8%), E-ocimene (1–10%), myrcene (8–45%) and terpinolene (0.12–22%).The monoterpene composition was useful to distinguish the monoecious cultivars from the dioecious ones. β-Caryophyllene (7–28%), α-humulene (3–12%), and caryophyllene oxide (2–6%) were the main sesquiterpenes. Tetrahydrocannabinol (THC) was present in traces in the EOs of only two dioecious cultivars cultivated in 2005. Cannabinol (CBN) was not detected in the essential oils, while the no-hallucinogenous cannabidiol (CBD) was found as typical volatile constituent in several analysed cultivars. These findings were also confirmed by the headspace GC–MS analysis carried out on the same samples. The analysed EOs obtained from fibre hemp varieties cultivated in Central Italy were characterized by an interesting and specific terpene composition with a legal and safe cannabinoid content. They were obtained from freshly plant inflorescences, which usually represent a waste material from C. sativa L. fibre varieties. The present study strengths the hypothesis to grow hemp as a multi-use crop through a complete utilization of the plant material using inflorescences to produce essential oils as natural flavour and fragrance additives.
The essential oil obtained by hydrodistillation of freshly harvested Indian Cannabis sativa L. was found to contain the following constituents that have not previously been reported: α-pinene, camphene, β-pinene, α-terpinene, β-phellandrene, γ-terpinene, linalool, trans-linalool oxide, sabinene hydrate, α-bergamotene, terpinene-4-ol, β-farnesene, α-terpineol, α-selinene, curcumene, and caryophyllene oxide. The presence of trace amounts of two alcohols and of an α,β-unsaturated ketone, for which gas chromatographic and spectral characteristics are recorded, was also detected.
Headspace Solid-phase micro extraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) has been used to isolate and identify the volatile compounds from the leaves of Cannabis sativa growing in Kashmir. The analysis led to the identification of 17 volatile components constituting 94.8 % of the total identified components. The chemical composition of the SPME extract from the leaves of C. sativa comprised mainly of sesquiterpene hydrocarbons (64.3%), monoterpene hydrocarbons (18.4%) and alcohols (10.3%). The major components identified in the HS-SPME extract were transcaryophyllene (36.9%), a-humulene (16.2%), a-pinene (10.7%), 3-hexen-1-ol-acetate (6.2%) and ?-pinene (4.2%). The current study is the first report involving rapid analysis of volatile components of C. sativa by HS-SPME.
When irradiated in the presence of photosensitizes, myrcene cyclizes to give 5,5-dimethyl-1-vinyl-bicyclo[2.1.1]hexane (1). 3-Methylene-1,5-hexadiene, 3-methylene-1,6-heptadiene, and 6-methyl-3-methylene-1,5-heptadiene cyclize under similar conditions to the corresponding bicyclo[2.1.1]hexanes. The geometric isomers of 3-methylene-1,5-heptadiene give identical mixtures of 5-methyl-2-methylenebicyclo[2,1,1]hexanes. The result indicates that cyclization involves formation of long-lived diradical intermediates. Preferential cyclization by way of intermediates containing five-membered rings is noted and discussed.
The introduction and development of comprehensive two-dimensional gas chromatography offers greatly enhanced resolution and identification of organic analytes in complex mixtures compared to any one-dimensional separation technique. Initially promoted by the need to resolve highly complex petroleum samples, the technique's enormous separation power and enhanced ability to gather information has rapidly attracted the attention of analysts from all scientific fields. In this Minireview, we highlight the fundamental theory, recent advances, and future trends in the instrumentation and application of comprehensive two-dimensional column separation.
R )—(+)‐Limonene was photooxidized in the presence of Rose Bengal as catalyst. After TLC isolation, the hydroperoxides formed were separated directly by HRGC and analyzed by MS (El; Cl). Each hydroperoxide isomer was then isolated by HPLC for structure determination which after reduction of the HOO group with sodium borohydride was performed by ¹ H‐NMR and ¹³ C‐NMR.
Six hydroperoxide isomers formed by oxidation of the endocyclic double bond were identified. The compounds eluted from the HRGC column in the following order (proportions are given in brackets) I (40.1%) (1 S , 4 R )‐ p ‐mentha‐2, 8‐diene 1‐hydroperoxide; II (5.8%) (1 R , 4 R )‐ p ‐mentha‐2, 8‐diene 1‐hydroperoxide; III (20.6%) (2 R , 4 R )‐ p ‐mentha‐[1(7), 8]‐diene 2‐hydroperoxide; IV (8.5%) (2 R , 4 R )‐ p ‐mentha‐6, 8‐diene 2‐hydroperoxide; V (4%) (2 S , 4 R )‐ p ‐mentha‐6, 8‐diene 2‐hydroperoxide; and VI (21.0%) (2 S , 4 R )‐ p ‐mentha‐[1(7), 8]‐diene 2‐hydroperoxide. Direct HRGC separation of the limonene hydroperoxides offers, inter alia , the possibility of determining their flavor qualities by HRGC/effluent sniffing.
The essential oils of five different cultivars of Cannabis sativa contained as main compounds α-pinene, myrcene, trans-β-ocimene, α-terpinolene, trans-caryophyllene and α-humulene. The content of α-terpinolene divided the cultivars in two distinct groups, an Eastern European group of cultivars of approximately 8% and a French group of cultivars of around 16%. Therefore, this compound might be suitable as a genetic marker for the two breeding centres for the fibre types of Cannabis sativa. The content of trans-caryophyllene was up to 19%. However, the content of caryophyllene oxide did not exceed 2%. The antimicrobial activity of the essential oil of Cannabis sativa can be regarded as modest. Nevertheless, cultivar differences were visible. Δ-9-tetrahydrocannabinol (THC) could not be detected in any of the essential oils and the amount of other cannabinoids was very poor. Copyright © 2001 John Wiley & Sons, Ltd.
The field of predictive microbiology is rapidly widening and ranges from bioreactor engineering to the modeling of foodstuff
degradation and contamination. In this chapter, we point out how important the choice of experimental conditions is. We also
briefly describe some tools (methodology of experimental research) that can be used to design optimal experimental strategies
with respect to the study's aim. Emphasis is given on the optimization of microbial metabolite production, with an example
showing the screening of factors and another illustrating the use of the response surface methodology.
Key WordsPredictive microbiology–methodology–experimental research–optimal design–response surface methodology–screening of factors–experimentation plan
To determine whether the terpenoid composition of the essential oil of Cannabis is useful for chemotaxonomic discrimination, extracts of pistillate inflorescences of 162 greenhouse-grown plants of diverse origin were analyzed by gas chromatography. Peak area ratios of 48 compounds were subjected to multivariate analysis and the results interpreted with respect to geographic origin and taxonomic affiliation. A canonical analysis in which the plants were pre-assigned to C. sativa or C. indica based on previous genetic, morphological, and chemotaxonomic studies resulted in 91% correct assignment of the plants to their pre-assigned species. A scatterplot on the first two principal component axes shows that plants of accessions from Afghanistan assigned to the wide-leaflet drug biotype (an infraspecific taxon of unspecified rank) of C. indica group apart from the other putative taxa. The essential oil of these plants usually had relatively high ratios of guaiol, isomers of eudesmol, and other unidentified compounds. Plants assigned to the narrow-leaflet drug biotype of C. indica tended to have relatively high ratios of trans-β-farnesene. Cultivars of the two drug biotypes may exhibit distinctive medicinal properties due to significant differences in terpenoid composition.
The photohydroperoxidation of the stereoisomers of three terpenes ((+) and (−) α-pinene, (+) and (−) β-pinene and (+) and (−) limonene) has been performed by using photocatalysts, such as zinc oxide, or sensitizers, such as anthracene or rose Bengal, supported on cross-linked polystyrene. Hydroperoxides accumulated alone in the first stages of sensitized oxidation but were always associated with alcoholic and carbonyl products in the case of ZnO. Secondary products obtained for longer exposure times in sensitized oxidations were identified by gas chromatography/mass spectrometry and mechanisms for their formation, deriving from the photolysis of parent hydroperoxides, were suggested.
The present study focused on inhibitory activity of freshly extracted essential oils from three legal (THC<0.2% w/v) hemp varieties (Carmagnola, Fibranova and Futura) on microbial growth. The effect of different sowing times on oil composition and biological activity was also evaluated. Essential oils were distilled and then characterized through the gas chromatography and gas chromatography-mass spectrometry. Thereafter, the oils were compared to standard reagents on a broad range inhibition of microbial growth via minimum inhibitory concentration (MIC) assay. Microbial strains were divided into three groups: i) Gram (+) bacteria, which regard to food-borne pathogens or gastrointestinal bacteria, ii) Gram (-) bacteria and iii) yeasts, both being involved in plant interactions. The results showed that essential oils of industrial hemp can significantly inhibit the microbial growth, to an extent depending on variety and sowing time. It can be concluded that essential oils of industrial hemp, especially those of Futura, may have interesting applications to control spoilage and food-borne pathogens and phytopathogens microorganisms.
The sweet odor of marihuana is of interest for instrumental monitoring of illicit drug traffic and for applications in forensic work. Headspace volatiles of marihuana and hashish of different origin are examined by gas chromatography, and relative compositions of 24 samples are compared. No correlation between volatile make up and geography was found, but the profiling procedures are shown to be of use in the forensic problem of relating samples to a common source.
In previous reports the presence of cannabinoids in the distilled essential oil of Cannabis sativa L. was proved, besides the presence of mono– and sesquiterpene hydrocarbons.
In this paper the localization of the cannabinoids in the glandular hairs of the leaves and with that the possible biogenetic relation with the components of the essential oil are demonstrated by microscopic examination after colouring tests and gaschromatographic analysis of the isolated contents of individual glandular hairs. Quantitative data about the relation between essential oil and cannabinoids are obtained by comparing the extracts without and after preceding steam distillation.
On acount of the origin of the seed (birdseed), special attention was paid to the botanical description of the plant material and to the counting of chromosomes.
THE sweet odour of Cannabis sativa, L. is of special interest as a possible means for the detection of illicit marijuana traffic. US Customs dogs, trained to alert to the odour of marijuana and hashish, are successfully used to find concealed contraband. An electronic ``sniffer'' that employs a portable quadrupole mass spectrometer to detect volatile vapours of drugs, including marijuana, has also been demonstrated1. Although the chemical composition of the essential oil of marijuana has been thoroughly investigated2-4, little is known about the specific composition of the emitted aroma constituents. The composition of this headspace, as it contributes directly to the odour, is more significant than that of the oil for the characterization of marijuana aroma5. Aroma profiles of fruits and vegetables have been obtained by direct gas chromatographic analysis of the headspace components6. We used this technique, which permits rapid determination of labile constituents under mild analytical conditions, to analyse marijuana aroma, and we report our preliminary results here.
The composition of the steam-distilled volatile oil of fresh and air-dried, indoor-grown marijuana was studied by GC/FID and GC/MS. In all, 68 components were detected of which 57 were fully identified. Drying of the plant material had no effect on the qualitative composition of the oil and did not affect the ability of individuals familiar with marijuana smell to recognize the odor.
Comprehensive two-dimensional gas chromatography (GC x GC) is a novel technique which is rapidly gaining importance for the analysis of complex samples. In the present review, attention is devoted to the principle and advantages, and main characteristics such as modulation, column combinations, detector requirements and data processing, of the technique. Specifically, GC x GC of a variety of real-life samples is discussed to demonstrate the applicability of the technique, with emphasis on the usefulness of the ordered-structure principle and on the analyte-identification power provided by a combination with time-of-flight mass spectrometric detection.
The cannabis plant (Cannabis sativa L.) and products thereof (such as marijuana, hashish and hash oil) have a long history of use both as a medicinal agent and intoxicant. Over the last few years there have been an active debate regarding the medicinal aspects of cannabis. Currently cannabis products are classified as Schedule I drugs under the Drug Enforcement Administration (DEA) Controlled Substances act, which means that the drug is only available for human use as an investigational drug. In addition to the social aspects of the use of the drug and its abuse potential, the issue of approving it as a medicine is further complicated by the complexity of the chemical make up of the plant. This manuscript discusses the chemical constituents of the plant with particular emphasis on the cannabinoids as the class of compounds responsible for the drug's psychological properties.
Although comprehensive two-dimensional gas chromatography (GC x GC) has been on the scene for more than 15 years, it is still generally considered a relatively novel technique and is yet far from being fully established. The revolutionary aspect of GC x GC, with respect to classical multidimensional chromatography, is that the entire sample is subjected to two distinct analytical separations. The resulting enhanced separating capacity makes this approach a prime choice when GC analysts are challenged with highly complex mixtures. The combination of a third mass spectrometric dimension to a GC x GC system generates the most powerful analytical tool today for volatile and semi-volatile analytes. The present review is focused on the rather brief, but not scant, history of comprehensive two-dimensional GC-MS: the first experiments were carried out at the end of the 1990s and, since then, the methodology has been increasingly studied and applied. Almost all GC x GC-MS applications have been carried out by using either a time-of-flight or quadrupole mass analyzer; significant experiments relative to a variety of research fields, as well as advantages and disadvantages of the MS systems employed, are discussed. The principles, practical and theoretical aspects, and the most significant developments of GC x GC are also described.
The Atlas of Spectral Data of Sesquiterpene Hydrocarbons
- D Joulain
- W Konig
D. Joulain, W. Konig, The Atlas of Spectral Data of Sesquiterpene Hydrocarbons, EB-Verlag, Hamburg, 1998.
Comprehensive Chromatography In Combination With Mass Spec-trometry
- L Mondello
L. Mondello, Comprehensive Chromatography In Combination With Mass Spec-trometry, John Wiley & Sons, Inc., 2011.
Composition of genuine indian Cannabis sativa
- E Stahl
- R Kunde
The essential oil of marihuana. Composition of genuine indian Cannabis sativa, Can. J. Chem.
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