G3M T typing by dot immunobinding with monoclonal anti-G3M T antibody

Dilutions of 100 serum samples of various GM phenotypes were dotted onto a nitrocellulose membrane. The serum dot-blots were detected with peroxidase-labeled anti-G3M T monoclonal antibody (anti-G3M T MAb). Up to a 1 : 256 dilution could be G3M T-typed correctly. By use of anti-G3M T MAb and peroxidase-labeled anti-mouse IgG or the biotin-avidin system instead of use of labeled anti-G3M T MAb, up to a 1 : 512 or 1 : 1024, respectively, dilution was typable. As with previous work with G3M G MAb, the dot immunobinding (DIB) method for G3M T typing was found very simple and practical.     

A 2‐Year Study of Δ 9‐tetrahydrocannabinol Concentrations in Drivers: Examining Driving and Field Sobriety Test Performance,,

From November 1, 2010 through November 30, 2012, 1204 whole‐blood samples were confirmed to contain THC alone or in combination with other drugs out of nearly 5000 Orange County, California, drivers suspected of driving under the influence of drugs. The goal of this study was to examine police reports and drug recognition expert evaluations of THC‐positive samples within this 2‐year time frame to determine whether there is a correlation between whole‐blood THC concentrations and field sobriety tests performance on DRE and non‐DRE evaluations. The FSTs prove to be sensitive to impairment by marijuana although as suspected, the findings of this study did not find a correlation between performance on field sobriety tests and the concentration of THC tested in whole‐blood samples. Driving behaviors were also examined and found to be similar to those seen in alcohol impairment. Future studies examining DRE findings are needed to confirm the results.     

Potency trends of delta9-THC and other cannabinoids in confiscated marijuana from 1980-1997

The analysis of 35,312 cannabis preparations confiscated in the USA over a period of 18 years for delta-9-tetrahydrocannabinol (delta9-THC) and other major cannabinoids is reported. Samples were identified as cannabis, hashish, or hash oil. Cannabis samples were further subdivided into marijuana (loose material, kilobricks and buds), sinsemilla, Thai sticks and ditchweed. The data showed that more than 82% of all confiscated samples were in the marijuana category for every year except 1980 (61%) and 1981 (75%). The potency (concentration of delta9-THC) of marijuana samples rose from less than 1.5% in 1980 to approximately 3.3% in 1983 and 1984, then fluctuated around 3% till 1992. Since 1992, the potency of confiscated marijuana samples has continuously risen, going from 3.1% in 1992 to 4.2% in 1997. The average concentration of delta9-THC in all cannabis samples showed a gradual rise from 3% in 1991 to 4.47% in 1997. Hashish and hash oil, on the other hand, showed no specific potency trends. Other major cannabinoids [cannabidiol (CBD), cannabinol (CBN), and cannabichromene (CBC)] showed no significant change in their concentration over the years.     

A simple DNA extraction method for marijuana samples used in amplified fragment length polymorphism (AFLP) analysis

As a first step in developing a molecular method for the individualization of marijuana samples, we evaluated a plant DNA extraction kit. The QIAGEN plant DNeasy method uses a spin column format for recovery of DNA and is effective for obtaining high molecular weight DNA from leaf, flower (bud), and seed samples of marijuana. The average DNA yield was 125-500 ng per 100 milligrams of fresh plant tissue. The recovered DNA was of polymerase chain reaction (PCR) quality as measured by the ability to generate reproducible amplified fragment length polymorphism (AFLP) profiles. AFLP is a technique used to create a DNA profile for plant varieties and is being applied to marijuana samples by the authors to link growers and distributors of clonal material. The QIAGEN plant DNeasy method was simple, efficient, and reproducible for processing small quantities of marijuana into DNA.     

Investigations into the hypothesis of transgenic cannabis

The unusual concentration of cannabinoids recently found in marijuana samples submitted to the forensic laboratory for chemical analysis prompted an investigation into whether genetic modifications have been made to the DNA of Cannabis sativa L. to increase its potency. Traditional methods for the detection of genetically modified organisms (GMO) were used to analyze herbal cannabis preparations. Our analyses support the hypothesis that marijuana samples submitted to forensic laboratories and characterized by an abnormal level of Δ(9)-THC are the product of breeding selection rather than of transgenic modifications. Further, this research has shown a risk of false positive results associated with the poor quality of the seized samples and probably due to the contamination by other transgenic vegetable products. On the other hand, based on these data, a conclusive distinction between the hypothesis of GMO plant contamination and the other of genetic modification of cannabis cannot be made requiring further studies on comparative chemical and genetic analyses to find out an explanation for the recently detected increased potency of cannabis.     

The Stable Isotope Ratios of Marijuana. II. Strontium Isotopes Relate to Geographic Origin

Abstract:  Effectively addressing marijuana trade is aided by understanding marijuana geographic sources. We analyzed the ⁸⁷Sr/⁸⁶Sr of marijuana samples grown in 79 counties across the United States to determine if a primary geologic signal is retained in marijuana, which could therefore be useful for geographic sourcing. The marijuana results were compared with modeled bedrock ⁸⁷Sr/⁸⁶Sr values based on ⁸⁷Rb decay rates and a generalized geologic map of the U.S.A. A significant correlation was observed between marijuana ⁸⁷Sr/⁸⁶Sr and modeled bedrock ⁸⁷Sr/⁸⁶Sr. Although values clustered near the 1:1 relationship, there was a predominance of positive anomalies, perhaps attributable to carbonate bedrock. A small number of negative anomalies were also observed, which were generally associated with granitic bedrocks. These results suggest that strontium isotopes in marijuana record the geographic origins of marijuana, and that refinement of the base strontium map (or strontium isoscape) and improved understanding of other strontium sources would be productive.     

用ELISA检测精液及精斑中精子抗原的研究

本实验应用单克隆抗人精子抗体和酶标记羊抗人精子抗体,采用ELISA方法确定精子抗原成份的存在。对10份新鲜精液,15份精斑进行了验测,其结果阳性率为100%。新鲜精液(精子数约10,000万个/ml)稀释100万倍,精斑浸出液稀释50万倍,均可出现阳性。对唾液斑、尿斑、乳汁斑、阴道斑、汗斑及输精管结扎的精液均为阴性。实验结果表明,本法检验精子抗原具有灵敏度高,特异性好的优点。     

A survey of the potency of Japanese illicit cannabis in fiscal year 2010

In recent years, increased 'cannabis potency', or Δ(9)-tetrahydrocannabinol (THC) content in cannabis products, has been reported in many countries. A survey of Japanese illicit cannabis was conducted from April 2010 to March 2011. In Japan, all cannabis evidence is transferred to the Minister of Health, Labour and Welfare after criminal trials. The evidence was observed at Narcotics Control Department offices in major 11 cities. The total number of cannabis samples observed was 9072, of which 6376 were marijuana. The marijuana seizures were further classified, and it was found that in terms of the number of samples, 65.2% of them were seedless buds, and by weight 73.0% of them were seedless buds. Seedless buds were supposed to be 'sinsemilla', a potent class of marijuana. THC, cannabinol (CBN) and cannabidiol (CBD) in marijuana seizures exceeding 1g were quantified. The number of samples analyzed was 1115. Many of them were shown to contain CBN, an oxidative product from THC. This was a sign of long-term storage of the cannabis and of the degradation of THC. Relatively fresh cannabis, defined by a CBN/THC ratio of less than or equal to 0.1, was chosen for analysis. Fresh seedless buds (335 samples) contained an average of 11.2% and a maximum of 22.6% THC. These values are comparable to those of 'high potency cannabis' as defined in previous studies. Thus, this study shows that highly potent cannabis products are distributed in Japan as in other countries.     

Profiles of delta 9-tetrahydrocannabinol metabolites in urine of marijuana users: preliminary observations by high performance liquid chromatography-radioimmunoassay

Metabolic profiles of 11-nor-9-carboxylic acid-delta 9-tetrahydrocannabinol (COOH-THC) and other THC metabolites were determined in an infrequent and a frequent marijuana user by high performance liquid chromatography-radioimmunoassay (HPLC-RIA). In the infrequent user, no unconjugated COOH-THC was detected in urine samples for the first 8 h following smoking, whereas this metabolite was detected in the urine samples from a frequent user. A metabolite was also detected in the frequent user, which was not present in the urine sample from the infrequent user.     

Detection of human seminal γ-glutamyl transpeptidase in stains using sandwich ELISA

A sensitive and specific sandwich ELISA for human seminal γ-glutamyl transpeptidase (γ-GTP) was developed using a combination of monoclonal antibodies, SG1 and SG3, which we produced. For semen identification in forensic samples, we modified the assay so as to be more sensitive and to establish efficient extracting conditions. After testing the extracting abilities of several detergents, CHAPS and deoxy-BIGCHAP were chosen as the solubilizer. Polystyrene beads coated with SG1 were incubated with samples extracted by the detergents, and further with biotinylated SG3, followed by peroxidase-labeled streptavidin. γ-GTP was detected only in seminal samples. The sensitivity of this assay was 0.01 ng/ml of seminal γ-GTP equivalent to 10⁷ times diluted semen, which was ten times as compared with the previous plate assay. No significant seminal γ-GTP was detected in other biological stains such as blood, saliva and vaginal smear. The extract of a 500 fold diluted seminal stain, 8 months old, showed the detection limit. Seminal γ-GTP was detectable even in 14-year-old stains.     

The effects of marijuana and alcohol usage on handwriting

In this study the handwriting of twelve individuals under the influence of marijuana, alcohol, and a combination of the two was compared with control samples. It was found that marijuana smoking in moderate amounts had less effect on the handwriting of the individuals than did alcohol. It was also found that when the two were combined the gross changes observed were probably due to the effect of the alcohol alone.     

Medical Marijuana Registries: A Painful Choice?

Though the medical use of marijuana is legal in thirty-three states, it remains illegal under the federal Controlled Substances Act. Any marijuana use can subject individuals to severe criminal and civil penalties under federal law. States that condition patient access and treatment on registration in a state database impose real risks on their citizens. Although many scholars have written about the tension between federal and state treatment of marijuana, this is the first article to examine marijuana patient registry privacy and fundamental rights issues. This article first reviews the relationship between marijuana use and patient treatment, with a focus on health-care and privacy rights under state and federal law. The article then explains how marijuana registries compare to broader patient registries, such as contagious disease and other medical condition patient registries, and the unique issues presented by marijuana patient registries. It then discusses the elevated risk to constitutional, privacy, and fundamental rights that may result if states do not carefully construct marijuana registries. The article concludes by proposing principles for how both states and dispensaries should approach marijuana registries in order to provide health benefits and avoid harm to patients.     

Validation of the 4‐aminophenol color test for the differentiation of marijuana‐type and hemp‐type cannabis

The analysis of cannabis plant material submitted to seized‐drug laboratories was significantly affected by the signing of the Agricultural Improvement Act of 2018, which defined hemp and removed it from the definition of marijuana in the Controlled Substances Act. As a result, field law enforcement personnel and forensic laboratories now are in need of implementing new protocols that can distinguish between marijuana‐type and hemp‐type cannabis. Colorimetric tests provide a cost‐effective and efficient manner to presumptively identify materials prior to submission to a laboratory for analysis. This work presents the validation of the 4‐aminophenol (4‐AP) color test and demonstrates its utility for discriminating between marijuana‐type and hemp‐type cannabis (i.e., typification). Validation studies included the testing of numerous cannabinoid reference materials, household herbs, previously characterized cannabis plant samples, and real‐case samples. The 4‐AP test reliably produces a pink result when the level of Δ⁹‐tetrahydrocannabinol (THC) is approximately three times lower than the level of cannabidiol (CBD). A blue result is generated when the level of THC is approximately three times higher than that of CBD. Inconclusive results are observed when the levels of THC and CBD are within a factor of three from each other, demonstrating the limitations of the test under those scenarios.     

First systematic evaluation of the potency of Cannabis sativa plants grown in Albania

Cannabis products (marijuana, hashish, cannabis oil) are the most frequently abused illegal substances worldwide. Delta-9-tetrahydrocannabinol (THC) is the main psychoactive component of Cannabis sativa plant, whereas cannabidiol (CBD) and cannabinol (CBN) are other major but no psychoactive constituents. Many studies have already been carried out on these compounds and chemical research was encouraged due to the legal implications concerning the misuse of marijuana. The aim of this study was to determine THC, CBD and CBN in a significant number of cannabis samples of Albanian origin, where cannabis is the most frequently used drug of abuse, in order to evaluate and classify them according to their cannabinoid composition. A GC-MS method was used, in order to assay cannabinoid content of hemp samples harvested at different maturation degree levels during the summer months and grown in different areas of Albania. This method can also be used for the determination of plant phenotype, the evaluation of psychoactive potency and the control of material quality. The highest cannabinoid concentrations were found in the flowers of cannabis. The THC concentrations in different locations of Albania ranged from 1.07 to 12.13%. The influence of environmental conditions on cannabinoid content is discussed. The cannabinoid content of cannabis plants were used for their profiling, and it was used for their classification, according to their geographical origin. The determined concentrations justify the fact that Albania is an area where cannabis is extensively cultivated for illegal purposes.     

Sourcing Brazilian marijuana by applying IRMS analysis to seized samples

The stable carbon and nitrogen isotopic ratios were measured in marijuana samples (Cannabis sativa L.) seized by the law enforcement officers in the three Brazilian production sites: Pernambuco and Bahia (the country's Northeast known as Marijuana Polygon), Pará (North or Amazon region) and Mato Grosso do Sul (Midwest). These regions are regarded as different with respect to climate and water availability, factors which impact upon the isotope fractionations of these elements within plants. It was possible to differentiate samples from the dry regions (Marijuana Polygon) from those from Mato Grosso do Sul and Pará, that present heavier rainfall. The results were in agreement with the climatic conditions of the suspected regions of origin and this demonstrates that seized samples can be used to identify the isotopic signatures of marijuana from the main producing regions in Brazil.     

电喷雾和大气压化学法对大麻酚类物质离子化的比较

目的比较电喷雾电离(ESI)和大气压化学电离(APCI)两种模式对大麻酚类物质的离子化效果。方法采用UFLC-(ESI/APCI)MS分析方法,分别考察使用ESI和APCI时雾化电压、雾化气流量、干燥气流量、加热块温度、解离管温度等参数变化对大麻酚类物质的影响规律,确定最优条件参数组合,并比较ESI和APCI对大麻酚类物质的离子化效果。结果对于0.5μg/mL大麻二酚、大麻酚和四氢大麻酚标准品,ESI峰高分别为215 006、143 051、216 944,信噪比41.74、49.88、42.12,峰面积日内标准偏差(RSD)<3.96%;APCI峰高分别为140 238、226 505、247 753,信噪比78.37、131.03、138.46,峰面积日内标准偏差(RSD)<11.98%。结论检测大麻样品时,ESI为首选,基质复杂时可以使用APCI作为ESI的补充手段。     

Detection of human proteins in buried blood using ELISA and monoclonal antibodies: towards the reliable species indentification of blood stains on buried material.

The survival of human proteins in blood stains on fragments of cloth buried in exposed soil was examined in a 15-month investigation carried out from September 1990 to December 1991. During this period there was a wide variety of weather conditions. Samples were exhumed at 4-weekly intervals for 16 weeks and finally at 65 weeks; extracts of the stains were tested for albumin and IgG using a highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) performed with monoclonal antibodies. Human albumin survived well throughout the 15 months of study, but IgG could be detected only in the 4- and 8-week samples. The reactions for IgG were weaker than those for albumin, although the method's sensitivity (10 ng) was the same for each protein. Appropriate buried and non-buried control experiments were carried out using cloth, either unstained or stained with human blood or animal sera; there was no cross-reactivity between human and the other species investigated and soil did not affect the assay; under laboratory conditions, IgG and albumin survived equally well. The system's versatility was illustrated by using monoclonal anti-bovine-albumin to detect specific albumin in the extracts of buried cloth which has been stained with bovine serum. It was concluded that ELISA performed with monoclonal antibodies could be of great value in identifying blood stains for forensic purposes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Passive exposure in detection of low blood and urine cannabinoid concentrations

Whenever small amounts of drugs are present in blood or urine samples, especially of substances that are preferentially smoked such as cannabinoids, the discrimination between active and passive inhalation may cause severe problems. The statement of a passive exposure by marijuana smoke has been scrutinized reviewing the literature. The pharmacokinetics of smoked marijuana as well as experimental data on cannabinoid concentrations in plasma and urine samples following passive exposure are summarized. As a conclusion it seems urgent to enlarge the existing data base.     

Determination of Herbicides Paraquat,Glyphosate, and Aminomethylphosphonic Acid in Marijuana Samples by Capillary Electrophoresis

In this work, two methods were developed to determine herbicides paraquat, glyphosate, and aminomethylphosphonic acid (AMPA) in marijuana samples by capillary electrophoresis. For paraquat analysis, sample was extracted with aqueous acetic acid solution and analyzed by capillary zone electrophoresis with direct UV detection. The running electrolyte was 50 mmol/L phosphate buffer (pH 2.50). For glyphosate and AMPA, indirect UV/VIS detection was used, as these substances do not present chromophoric groups. Samples were extracted with 5 mmol/L hydrochloric acid. The running electrolyte was 10 mmol/L gallic acid, 6 mmol/L TRIS, and 0.1 mmol/L CTAB (pH = 4.7). The methods presented good linearity, precision, accuracy, and recovery. Paraquat was detected in 12 samples (n = 130), ranging from 0.01 to 25.1 mg/g. Three samples were positive for glyphosate (0.15–0.75 mg/g), and one sample presented AMPA as well. Experimental studies are suggested to evaluate the risks of these concentrations to marijuana user.     

Trace elemental analysis of drugs of abuse using synchrotron radiation total reflection X-ray fluorescence analysis (SR-TXRF)

Synchrotron radiation total reflection X-ray fluorescence spectroscopy (SR-TXRF) was utilized to analyze various trace elements in small amounts of drugs of abuse. Sample amounts of 1 microL solutions containing 10 microg of drugs (methamphetamine, amphetamine, 3,4-methylenedioxymethamphetamine, cocaine, and heroin) were spotted on silicon wafers for direct analysis. In addition, a leaflet of marijuana was set directly on a silicon wafer, and opium in the form of a soft lump was smeared on another silicon wafer for analysis. In these experiments, about 10 pg of contaminant elements could be detected. For example, in a seized methamphetamine sample, iodine was found, which could be indicative of synthetic route. In seized 3,4-methylenedioxymethamphetamine samples, variable amounts of phosphorus, calcium, sulfur, and potassium were found, which could not be detected in a control 3,4-methylenedioxymethamphetamine sample. For marijuana and opium, two spectral patterns were obtained that were far different from each other and could be easily discriminated. Using SR-TXRF, pg amounts of each trace element in 10 microg of various drugs can be easily detected, which is not the case either for a standard TXRF experimental system or for other elemental analysis techniques.