Hashish Samples Research Articles

Phytocannabinoid profile and potency of cannabis resin (hashish) of northwest Himalayas of India.

Cannabis is one of the most consumed illicit drugs and the potency of cannabis products is of note due to health-related concerns. Hand-rubbed hashish is the ancient technique of extracting psychoactive resin from cannabis plants and is practiced in the Indian Himalayas. This study establishes the cannabinoid profile and potency of hand-rubbed hashish collected from 20 regions of the northwest Himalayas. Fifty-eight hashish samples were analyzed using a validated high-performance liquid chromatography-diode array detection (HPLC-DAD) method. Ten cannabinoids were quantified including acidic (THCA & CBDA), and neutral compounds (CBDV, THCV, CBD, CBG, CBN, Δ9-THC, Δ8-THC, and CBC). The mean concentration (w/w%) of Δ9-THC is 26%; THCA is 15% and THCTotal is 40% is observed in the studied hashish samples. The majority (70%) of the hashish samples were categorized in chemotype I with the THC:CBD:CBN ratio of 91:3:4, and the remaining 30% were categorized under chemotype II with the ratio of 76:15:8. Diverse qualities of hashish are produced in the studied regions as per the seed, plant selection, and skills of manual rubbing, which results in potency variations. The average difference between the least and highest potent hand-rubbed hashish of a region is 27 w/w% (THCTotal). The other studied non-psychoactive cannabinoids have a mean w/w% of <5%, followed by 6% of CBDA. It is concluded that the cultivated and wild cannabis fields in the northwest Himalayas belong to the drug-type cannabis subspecies. Hand-rubbed hashish holds traditional significance and impacts the current policies of legislation.

Trends in Illicit Cannabis Potency based on the Analysis of Law Enforcement Seizures in the Southern Area of Rome.

Cannabis remains the most illicitly produced and consumed substance worldwide, and the average trans-Δ9-tetrahydrocannabinol (THC) content in cannabis products (marijuana, hashish) has increased over time. This paper presents data about THC concentration in cannabis resin samples seized by law enforcement from 2015 to 2022 in the southern area of Rome (Italy). From 2015 to 2022, more than 1000 hashish samples were analyzed; the average THC content was 18.0% and dramatically increased from 13.7% (2015) to 27.1% (2022). The potency of THC in some samples characterized by unusual shape and color was higher than 24% and, in a few cases, higher than 40%. The age group most involved in seizures of cannabis resin concerned males aged between 15 and 36 years old. The spread of this phenomenon increases the risk of adverse health outcomes. Many observational studies compare the increased cannabis potency with the onset of psychosis, depression, anxiety and cannabis use disorders (CUDs), mainly in young adults. THC-potency monitoring provides data that can be helpful to create a network of communication and interaction between universities, and legislative and public health institutions to support education, awareness and surveillance related to cannabis abuse.

Hashish seizures in Algeria over the 2019-2020 period: The rise of potent hashish hybrids.

In Algeria, hashish is by far the most common illicit drug. This study explores Algerian hashish over a two-year period (2019-2020). A total of 2583 hashish samples from 1707 seizures for a total quantity of 108 tons were analyzed using a validated high-performance liquid chromatography-diode array detection method (HPLC-DAD). The yearly arithmetic mean tetrahydrocannabinol (THC) concentration shows relative stability-18.67% in 2019 and 19.03% in 2020 with an overall mean THC content of 18.87% and standard deviation of 10.99%. High-potency hashish (THC content>20%) is by far the most predominant type, representing almost 50% of the total Algerian hashish seizures (mean and median around 29%). The overall mean of cannabidiol (CBD) was 2.45%, and 12% of the total studied seizures were of very low CBD concentration (CBD content<1%). Three distinct hashish chemotypes were identified: Chemotype I described the traditional Moroccan hashish with THC content ranging from 0% to 16%, Chemotype II hashish included most of the seizures and characterized by THC content ranging from 16% to 30%, and Chemotype III was characterized by hashish potency higher than 30% and very low CBD content. The identified chemotypes I, II, and III were characterized in a ternary plot, and the relative contents (THC:CBD:CBN) were about 67%:29%:4%, 88%:9%:3%, and 96%:2%:2%, respectively.

REDGIM COMO APLICATIVO DE SMARTPHONE PARA APLICAÇÕES QUIMIOMÉTRICAS POR MEIO DE ANÁLISE DE IMAGENS: UM USO EM PLS

REDGIM AS A SMARTPHONE APP FOR CHEMOMETRICS APPLICATIONS THROUGH IMAGE ANALYSIS: A USE IN PLS. The study of the spectrum of light has increased its range of action in the field of chemistry, with the help of several advanced equipment and innovative techniques that have become a strong ally of analytical chemistry. However, the simplicity of obtaining results integrated with mobile devices appears as an interesting path to be taken in the field of chemical analysis. This work addresses the dissemination of an application called REDGIM for smartphones on the Android platform capable of working with multivariate image analysis using chemometric methods through the luminous intensities of photos and R (red), G (green) and B (blue) channels. In order to show the potential of REDGIM, we propose two applications using the partial least squares (PLS) regression method: 1) quantification of free chlorine in tap water; and 2) quantification of cannabinol, (CBN, a type of cannabinoid) in marijuana and hashish samples. The free chlorine concentration obtained for two investigated samples was 0.73 and 0.74 mg L–1 and 0.74 and 0.76 mg L–1 by the spectrophotometer and by the application (container: cuvette), respectively. For the quantification of CBN in marijuana and hashish samples, the mean concentration ranged from 6.137 to 8.823 mg mL–1. The results obtained by the application are compatible with results from traditional techniques, such as UV-Vis spectrophotometry, for example. Therefore, REDGIM was proved to be a simple and accessible technological tool for the scientific community, being viable for chemical tests of a multivariate nature.

Characterization of Algerian‐Seized Hashish Over Eight Years (2011–2018). Part II: Chemical Categorization

In Algeria, large quantities of hashish are seized every year. This study aimed to investigate the total content of major cannabinoids in the illicit seized hashish in Algeria over an 8-year period (2011-2018) in order to establish the chemical profile of North African hashish. A total of 3265 hashish samples were analyzed using a validated high-performance liquid chromatography-diode array detection (HPLC-DAD) method, allowing the simultaneous quantification of both the acidic and the neutral forms of Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN). The results revealed a slight upward trend in the mean THC content, from 7.0% in 2011 to 9.4% in 2018, with an overall mean value of 8.4%. The overall means of CBD and CBN content were 3.5% and 0.8%, respectively. The number of high-potency hashish samples gradually increased to reach 6% in 2018. Two distinct hashish chemotypes were identified: the highly populated chemotype II, corresponding to the traditional medium-potency hashish ([THC+CBN]/CBD~2.16), and chemotype I, containing hashish samples of relatively high THC levels and low levels of CBD (ratio~4.90). Both chemotypes I and II were characterized in the ternary plot, and the proportions (THC:CBD:CBN) were about 85%:13%:2% and 60%:35%:5%, respectively.

Quantitative determination of CBD and THC and their acid precursors in confiscated cannabis samples by HPLC-DAD

Analysis of cannabis has gained new importance worldwide, mainly for quality control within the legalized recreational and medical cannabis industry, but also for forensic differentiation between drug-type cannabis and legal products such as fiber hemp and CBD-rich/THC-poor cannabis. We herein present an HPLC-DAD method for quantitative analysis of major neutral and acidic cannabinoids in herbal cannabis and hashish: Δ9-tetrahydrocannabinol (THC), Δ9-tetrahydrocannabinolic acid A (THCA), cannabidiol (CBD), cannabidiolic acid (CBDA), and cannabinol (CBN). Plant material was dried, homogenized and extracted with a mixture of methanol/hexane. Chromatographic separation of the analytes was achieved on a core-shell C8 column using gradient elution with water/acetonitrile containing 0.1% formic acid. The analytical run time was 13 min and analytes were detected at 210 nm. The method is selective, sensitive, accurate, and precise, as confirmed through validation according to ICH and AOAC guidelines. Linearity in herbal cannabis ranged from 0.04 to 4.00% for the neutral cannabinoids, and from 0.40 to 20% for the acids. Linear ranges in hashish samples were 0.13–13.33% and 1.33–66.66%, respectively. The presented method was successfully applied to characterize 110 cannabis samples seized by the Swiss police, demonstrating its applicability for routine cannabis potency testing in the forensic setting.

Cannabis resin in the region of Madrid: Adulteration and contamination

The aim of this study is to analyze the adulteration and contamination of cannabis resin obtained on the streets of Madrid, in order to establish whether it is suitable for human consumption. A total of 90 samples obtained through street vending in the Region of Madrid (CAM) were analyzed. Our results showed a direct relationship between the shape of the samples (acorn or ingot) and the presence of foreign elements, adulterants and microbiological contamination. Foreign elements were found in 64.7% of the ingot-shaped samples and in 30.2% of the acorn-shaped samples (p < 0.01); 25% of the samples were deliberately adulterated, 66.7% of which had an ingot shape. With regard to microbiological contamination, 93% of acorns were contaminated by E. coli, compared to 29.4% of ingots (p < 0.0001). In addition, all samples with fecal odor were acorns and were contaminated by E. coli. Ten per cent of the samples were contaminated by Aspergillus; of these, 66.7% had the shape of an acorn. Overall, our results showed that most (88.3%) of the hashish samples were not suitable for consumption. This percentage was significantly higher (p < 0.0001) in acorn than in ingot samples (100% vs. 58.8%). Hence, illegal street vending of hashish constitutes a public health issue.

VOCs as fingerprints for the chemical profiling of hashish samples analyzed by HS-SPME/GC–MS and multivariate statistical tools

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. 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. 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. 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.

The main cannabinoids content in hashish samples seized in Israel and Czech Republic

Cannabis, both herbal and resin, has been the most popular illicit drug in Israel in recent years. Until 2005, the main sources of cannabis resin (known also as hashish) to the Israeli drug market were Lebanon and India. Hashish from these sources can be distinguished by its external appearance. The aim of this study was to determine if there is any difference in the quality of the hashish from each source. For this purpose, we quantified the main cannabinoids, cannabidiol (CBD), Δ9-tetrahydrocannabinol (Δ9-THC), and cannabinol (CBN) of hashish from different police seizures of known origins, Lebanon, India and Morocco, that had been submitted to the Analytical Chemistry Laboratory of the Division of Identification and Forensic Science at the Israeli National Police Headquarters and subsequently to the Hebrew University for quantitative analysis. The results, based on many different seizures, showed that the average CBD content of hashish from Lebanon varied from 5.69% to 12.79% (an average 8.98 ± 0.59%), THC of hashish from Lebanon varied from 0.93% to 4.20% (an average of 2.38 ± 0.27 %), CBD of hashish from Morocco varied from 1.52% to 5.14% (an average of 3.72 ± 0.19%), THC of hashish from Morocco varied from 5.08% to 13.41% (an average of 9.21 ± 0.40%), CBD of hashish from India varied from 0.78% to 13.13% (an average of 4.59 ± 1.07%), and THC of hashish from India varied from 0.53% to 16.45% (an average of 6.35 ± 1.50%), At the same time, several other cannabinoids present in the samples in lower amounts were identified (cannabidivarol, CBDV; cannabicitran; Δ9-tetrahydrocannabivarol, Δ9-THCV; cannabivarol, CBV; cannabicyclol, CBL; cannabichromene, CBC; cannabielsoin, CBE; Δ8-THC; and cannabigerol, CBG). The samples, predominantly from Lebanon, Morocco, and India, were evaluated for chemical phenotype (drug type and fiber type) to determine the geographical origin of these samples.Indian hashish, seized by Czech Republic authorities, was analyzed qualitatively for many cannabinoids and terpenes. In this sample three new cannabinoids were identified: cannabiorcochromene, cis-Δ9-tetrahydrocannabivarol, and cannabinerol.

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.

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.

Cannabinoid findings in children hair - what do they really tell us? An assessment in the light of three different analytical methods with focus on interpretation of Δ9-tetrahydrocannabinolic acid A concentrations.

Hair analysis for drugs and drugs of abuse is increasingly applied in child protection cases. To determine the potential risk to a child living in a household where drugs are consumed, not only can the hair of the parents be analyzed but also the hair of the child. In the case of hair analysis for cannabinoids, the differentiation between external contamination and systemic uptake is particularly difficult, since the drug is quite often handled extensively prior to consumption (e.g. when preparing a joint) and smoke causes a further risk for an external contamination. Δ9-tetrahydrocannabinolic acid A (THCA-A), the non-psychoactive biogenetic precursor of Δ9-tetrahydrocannabinol (THC), is a suitable marker for external contamination since it is not incorporated into the hair matrix through the bloodstream in relevant amounts. In the presented study, hair samples from 41 children, 4 teenagers, and 34 drug-consuming parents were analyzed for THCA-A, THC and cannabinol (CBN) applying methanolic extraction and a fully validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method (Method 1). For comparison, a part of the samples was also analyzed applying alkaline hydrolysis followed by liquid/liquid extraction and gas chromatography-mass spectrometry (GC-M)S (Method 2), or by headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) (Method 3). Furthermore, 458 seized marihuana samples and 180 seized hashish samples were analyzed for the same cannabinoids by gas-chromatography-flame ionization detector (GC-FID). In all but one of the hair samples, the concentration of THCA-A was higher than the concentration of THC and in 14 cases no THC could be detected despite the presence of THCA-A, suggesting that in almost all cases a significant external contamination had occurred. Within-family comparison showed a higher THCA-A/THC ratio in hair of children than of their consuming caregivers. Mean and median of this ratio of all hair samples (6.7 and 4.2) were between those of marihuana (11.0 and 8.3) and hashish (2.8 and 2.1) with a large variation in all samples. Comparison of the Methods 1 to 3 showed clearly that the choice of the analytical procedure has a strong influence on the quantitative results, mainly because of decarboxylation of THCA-A during hair hydrolysis by NaOH and other analytical steps, which lead to artifactually elevated THC concentrations. In conclusion, these findings suggest that the major part of the cannabinoids detected in the hair samples from children arose from an external contamination through 'passive' transfer by e.g. contaminated hands or surfaces and not from inhalation or deposition of side stream smoke.

Interference of alkanes in the gas chromatographic analysis of cannabis products.

Abstract Leaves of young marihuana plants (Cannabis sativa L.) were found by gas liquid chromatography to contain appreciable amounts of two long-chain alkanes, n-heptacosane and n-nonacosane. These alkanes, along with other straight chain alkanes ranging from C19 to C32 could also be detected as minor components in a variety of other marihuana and hashish samples. Depending on the polarity of the g.l.c. column used, the alkanes may have retention times similar to those of the major cannabinoids and thus interfere with qualitative and quantitative analyses of the latter. The findings indicate that g.l.c. alone cannot be used as an accurate and reliable technique for cannabinoid analysis, unless the alkanes are previously removed. However, the alkane composition may be of additional advantage in determining the origin of seized cannabis samples.

Cannabinoid contents in cannabis products seized in São Paulo, Brazil, 2006–2007

A rapid and simple method was optimized for determination of Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN) contents in cannabis products by gas chromatography with flame-ionization detection (GC-FID), using diazepam as internal standard. All parameters of validation of the method such as linearity, intraassay precision, and limits of detection and quantification of the analytes were satisfactory. Using the described method, cannabinoid contents of 55 cannabis product samples seized in Sao Paulo City, Brazil, in 2006 and 2007 were measured. Δ9-THC content in marijuana and hashish samples varied between 0.08% and 5.5%, with an average of 2.5%. The phenotypic ratio showed that the products were able to be designated as “drug type.”

GC‐MS Analysis of Hashish Samples: A Case of Adulteration with Colophony

This report describes the analytical characterization of 16 hashish samples confiscated in Italy. The samples were solvent extracted and subjected to GC-MS analysis for the separation and quantitation of the main cannabinoids. One of the analyzed samples was shown to contain extraneous compounds, identified as resin acids characteristic of colophony (rosin). Colophony is a natural resinous product obtained from various species of pine, spruce, and larch; it is a skin sensitizer and its fumes produce nonspecific irritation that can cause bronchial asthma. Similar adulterations of hashish have not been reported previously; therefore, at present there is no information about the effects of smoking colophony or a combination of hashish and colophony. It is presumed, however, that such a combination would be highly detrimental to the health of the user.

Analytical characterisation of hashish samples

This report describes the analytical characterisation of two hashish samples to establish if they belong to a common lot, although their physical appearance is quite different. The samples were extracted and subjected to HPLC and GC for the separation and quantitation of the main cannabinoids. GC-MS enabled the selective identification of minor cannabinoids and terpenes. Hashish samples were also mineralised and analysed for metal traces by atomic absorption spectrometry (AA).

Simultaneous separation and identification of hashish constituents by coupled liquid chromatography-mass spectrometry (HPLC-MS)

This paper reports the use of liquid chromatography for the separation and determination of the major cannabinoids extracted from hashish samples. The direct coupling to the mass spectrometer enables the selective identification both of neutral and acidic cannabinoids. The developed method does not require any preliminary derivatization and should, therefore, be of interest in forensic analysis for simple and unequivocal determination of hashish constituents.

Sample preparation by supercritical fluid extraction for quantification a model based on the diffusion-layer theory for determination of extraction time

A mathematical model based on the diffusion-layer theory was elaborated in order to calculate the extraction time in dynamic supercritical fluid extraction required to reach a predefined level of extraction recovery. The goodness of the model is demonstrated by application to the extraction of the main neutral cannabinoids from marihuana and hashish samples. For monitoring of the cannabinoid content of extracts normal-phase HPLC was applied. To obtain reliable quantitative results, the extraction time ensuring a predefined level of recovery should be calculated for each individual sample according to the model because the extraction recovery depends on the sample matrix. The systematic error caused by the unextracted compounds can be eliminated by correction of the experimental data. For semi-quantitative determinations, where a knowledge of the correct value of the extraction recovery is not important, as a rule of thumb the extraction of marihuana with carbon dioxide of density 0.9 g/ml at 40°C for 34 min and of hashish for 18 min can be suggested. The application of the proposed extraction times ensured at least a 95% recovery for the main neutral cannabinoids.

Investigation of Hashish: Content Uniformity of Different Samples by Coupled HPLC/PC‐Analysis

AbstractStandardized storage experiments with hashish and its isolated pure components have shown that for the judgement of hashish samples a comparison of total values of CBD/CBDA, CBN/CBNA and THC/THCA is important. The investigation is carried out by HPLC and is illustrated by a histogram obtained with the aid of a simple data processing program. Changes in the individual cannabinoid contents resulting through different storage conditions are largely compensated for. These results should be confirmed through an additional comparison with standardized HPLCs calculated on the basis of the same weight‐in quantity. This is demonstrated on five different confiscated hashish samples.

Analysis of the ageing processes in hashish samples from different geographic origin

Hashish samples from Morocco, Lebanon, Iran, India, Pakistan, Afghanistan have been stored either in darkness or in daylight at 18–22°C and analysed every 20–30 days for a period of 22 months. Each of the samples was analysed by gas-chromatography with α-cholestane as the internal standard. As a result of this study we have observed distinctive ageing processes due to the physical and chemical peculiarity of samples of different geographic origin. The information obtained from this work may have a forensic application in the illicit traffic of hashish.

Cannabinoid Content of Fifty Seized Hashish Samples

Abstract During the last few years, police seizures of hashish have increased rapidly in Sweden. A large part of the seized samples have been sent to this laboratory for analysis, and the case load has risen accordingly (Table 1).