Giovanna Tassoni

Determination of primary active constituents in Cannabis preparations by high-resolution gas chromatography/flame ionization detection and high-performance liquid chromatography/UV detection

Abstract For a complete quantitative analysis of primary active constituents in Cannabis preparations Δ 9 -tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN), we have compared two different chromatographic techniques, high-resolution gas chromatography (HRGC)/flame ionization detection (FID) and high-performance liquid chromatography (HPLC)/UV. The two different methods have been validated using crude drug (hashish) with methyloleate and tetraphenylethylene as internal standard for HRGC/FID and HPLC/UV, respectively.

Hair analysis in order to evaluate drug abuse in driver's license regranting procedures.

In Italy, driving under the influence of drugs determines the suspension of the offenders drivers license. To regain the license the person must be drug free during an observation period. People whose license has been revoked or suspended can obtain, or re-obtain their drivers license subject to the judgment of a medical commission. The exclusion of illicit drug use is determined by means of toxicological analysis, mainly on urine or hair matrices. We reported the results of several years of experience of the forensic toxicology laboratory of the University of Macerata in the use of hair analysis for the assessment of past exposure to drugs in people suspected of driving under the influence of drugs. From 2004 to 2013, 8612 hair samples, were analyzed for opiates, cocaine and delta-9-tetrahydrocannabinol (Δ(9)-THC) using gas chromatography/mass spectrometry (GC/MS) method. We used a cutoff (SoHT or national guidelines) to determine the positive data, regardless of the hair sample concentrations. 1213 samples resulted positive, 71.7% were positive for cocaine and metabolites, 19.8% for morphine and metabolites, 8.5% for Δ(9)-THC. We also studied the timeframe of the abuse, as well as gender and age distribution of positive subjects. Moreover, we analyzed the possible deterrent effect of the hair analysis on driving under the influence of psychoactive substances.

Bile Analysis in Heroin Overdose

Abstract:  Following its metabolism in the liver, morphine and its metabolites can be directly eliminated in bile. Then, they undergo the enterohepatic cycle (EHC) and mostly reappear in the circulation. We report a case showing the presence of morphine in bile (21.3 μg/mL) and hair (4.8 ng/mg) but not in blood, urine or the liver of an addict who survived in hospital for about 144 h (6 days). These data would indicate that the EHC does not play any role about 144 h after the last injection, and directly confirms that gall bladder is a storage depot for morphine. They constitute the first report of a demonstration of the effect of the EHC on morphine bioavailability in an addict, and could be considered as indication, without supporting circumstantial evidence, that the morphine level in bile is related to chronic opiate use.

Detection of Cannabinoids by ELISA and GC–MS Methods in a Hair Sample Previously Used to Detect Other Drugs of Abuse

A forensic standard procedure is described that combines enzyme-linked immunoassay for screening and gas chromatography-mass spectrometry (GC-MS) for confirmation to detect drugs of abuse in a sample before used to detect opioids and cocaine. We used two equal aliquots of the same previously selected cannabinoid positive hair samples, one of which was subjected to acid hydrolysis. Afterward, both the aliquots were subjected to basic extraction and then to immunoassay screening. After derivatization, the GC-MS parameters were the same for both the aliquots for the determination of the cannabinoids (Δ(9)-tetrahydrocannabinol, cannabidiol and cannabinol). The results show that there were no statistical differences between the nonpreviously treated and the pretreated hair samples for the quantification of the three cannabis products for immunochemical procedure. No differences between the two groups were shown as for GC-MS confirmation procedures. All substances showed a good linearity between 0.05 and 2 ng/mg. The limit of detection ranged from 0.02 to 0.03 ng/mg, and the limit of quantification was 0.05 ng/mg for all substances. To our knowledge, this is the first time that screening and confirmation procedures have been applied on the same sample of hair to detect more than one drug of abuse.

Driving under the effect of drugs: Hair analysis in order to evaluate recidivism.

Under Italian law drug addiction and regular drug abuse are incompatible with driving ability. One important problem with the enforcement of the impaired driving law is the large number of people that re-offend. To regain their license, offenders must be drug-free for the duration of an observation period, according to the judgement of a medical commission. The exclusion of illicit drug use is determined by toxicological analysis. A few studies exist that have used a hair matrix to monitor recidivism. Hair is an attractive matrix for monitoring drug recidivism, due to the large time window for drug detection, and to the non-alterability of this matrix. We report the results of several years of experience at our forensic toxicology laboratory in the use of hair analysis for the assessment of past exposure to drugs in persons suspected of driving under the influence of drugs. 5592 subjects were analyzed for opiates, cocaine and delta-9-tetrahydrocannabinol (Δ9-THC) using a GC/MS method. 1062 (19.0%) subjects resulted positive. From this group, the individuals that resulted positive at least at the second control were considered recidivists (243, 22.9%). 79.7% of recidivist subjects were positive for cocaine and metabolites, 14.9% for morphine and metabolites, 5.4% for Δ9-THC. We also studied the time frame of the abuse, as well as gender and age distribution of recidivist subjects. Furthermore, we analyzed risk factors associated with recidivist behaviour. Our results show that cocaine consumption was the only factor that showed significance with regard to increased likelihood of being a recidivist.

Toxicological Analysis of Opiates from Alternative Matrices Collected from an Exhumed Body

In this case study, the body of a 45‐year‐old man was exhumed after 1 year at the request of the public prosecutor to assess whether the death was caused by drug consumption. Toxicological analyses were performed on several matrices, including liver, kidney, and the alternative matrices hair and teeth. The systematic toxicological analysis (STA), which consisted of basic and acid liquid/liquid extraction and gas chromatography–mass spectrometry (GC‐MS) analysis, showed the presence of opiates in each of the matrices analyzed. Subsequently, to confirm and quantify the presence of opioids, samples of each of the matrices were subjected to solid‐phase extraction and specific GC‐MS analysis. The case presented demonstrates the possibility of drug detection in an exhumed body that has been buried for 1 year, despite the problems of quantitative interpretation of the data, and that toxicological results could be useful along with other forensic evidence.