Claudia Trignano

Cocaine and heroin in waste water plants: a 1-year study in the city of Florence, Italy

The diffusion and trends in use of each substance is a basic information in policy planning of strategies aiming at deterrence of drug abuse or in the organization of the fight against drug trafficking. The actual diffusion of illicit drugs in a population is hardly measurable, but, among the various measures available, the analysis of waste water plants represents one of the most reliable source of data. We analyzed waste water in order to monitor illicit drug use by local population. We investigated the use of cocaine and heroin in the city of Florence, Italy, over a 1-year (July 2006-June 2007) period using state-of-the-art measuring techniques from waste water samples. Cocaine, benzoylecgonine, and morphine were determined in water samples by gas chromatography-mass spectrometer, and the amount of illicit substance was estimated. Data indicate for cocaine a bimodal distribution (December and March), while heroin showed a main peak in April. The heroin-to-cocaine use ratio in terms of estimated doses per month ranged from 0.11 to 0.76, representing new evidence of wider distribution of cocaine than heroin in Florence. Waste water analysis can become a valuable tool in monitoring use of illicit drugs over time. In particular, it can highlight changes in the magnitude and relative use of illicit drug at a population level thereby becoming useful to develop strategies against drug trafficking and abuse. If routinely performed, it can be part of Epidemiologic Surveillance Programmes on drug abuse.

What constitutes a normal ante-mortem urine GHB concentration?

Gamma-hydroxybutyric acid (GHB) is endogenously produced within the central nervous system, however it is also used as a medication for the treatment of a variety of clinical conditions, sold under the name Zyrem in the United States and Alcover in Europe. It is a very dangerous drug with a very limited safety margin, and is classified as a controlled substance in many countries. The interpretation of post-mortem studies of GHB concentrations is problematic; GHB can be detected in urine and blood from non-GHB users, both before and after death, and concentrations in both matrices may rise with prolonged storage. Because it is produced as a post-mortem artifact, forensically defensible cut-offs for post-mortem blood concentrations have yet to be established. Given the enormous degree of inter and intra-individual variation in GHB production that has been documented, it is unlikely they ever will. The important issue for forensic scientists is whether the detection of GHB in urine, in concentrations above some yet to be determined value, can be used as evidence for drug facilitated assault. In an attempt to see if a cut-off level could be determined we analyzed urine from 39 alcoholics who were being treated with known oral doses of Alcover (group 1), and compared the results with concentrations found in the urine of 30 volunteers who had no exogenous GHB intake (group 2), and 30 urine specimens taken from the alcoholics before they initiated GHB therapy (Alcover treatment group 3). More than one third (36.6%) of subjects being treated with GHB were found to have urinary GHB concentration that fell between 2.75 and 10 microg/mL. The data suggests that caution must be used when applying the currently used cut-off of 10 microg/mL.

Cross-reactivities of 41 new amphetamine designer drugs to EMIT® immunoassays

Amphetamine designer drugs are central nervous system stimulants, and are widely diffused in illegal markets. Monitoring of drugs of abuse in biological fluids is successfully used for clinical and forensic applications. In particular, the urine matrix allows the verification of drug intake in the short and medium term. In a forensic toxicology laboratory, typical analysis for these drugs involves an immunoassay screening method. Here we describe the cross-reactivity profiles of 41 new amphetamine designer drugs to the urine drug tests EMIT® II Plus (Amphetamines assay and Ecstasy assay).

Validated LC–MS-MS Method for Multiresidual Analysis of 13 Illicit Phenethylamines in Amniotic Fluid

A multi-residue analytical method was developed for the determination in amniotic fluid (AF) of 13 illicit phenethylamines, including 12 compounds never investigated in this matrix before. Samples were subject to solid-phase extraction using; hydrophilic-lipophilic balance cartridges which gave good recoveries and low matrix effects on analysis of the extracts. The quantification was performed by liquid chromatography electrospray tandem mass spectrometry. The water-acetonitrile mobile phase containing 0.1% formic acid, used with a C18 reversed phase column, provided adequate separation, resolution and signal-to-noise ratio for the analytes and the internal standard. The final optimized method was validated according to international guidelines. A monitoring campaign to assess fetal exposure to these 13 substances of abuse has been performed on AF test samples obtained from pregnant women. All mothers (n = 194) reported no use of drugs of abuse during pregnancy, and this was confirmed by the analytical data.

LC-MS/MS analysis of acetaminophen and caffeine in amniotic fluid

The intake of several substances by pregnant women could be hazardous to the fetus and mothers health: many substances can cross the placenta and reach the fetal compartment, causing adverse outcomes. Consequently, to accurately measure the presence of xenobiotics in fetal matrices, sensitive and specific bioanalytical methods are necessary: this would allow the assessment of fetal exposure to substances which, although licit, can be dangerous for the fetal and childs growth. The aim of this study was to develop and validate a liquid chromatography tandem mass spectrometry method for the simultaneous determination and quantitation of caffeine and acetaminophen in amniotic fluid. Amniotic fluid is a quite complex biological matrix and, as such, it requires a purification step prior to analysis. The extraction method has been optimized by comparing three different commercially available SPE cartridges (Supel™ Select HLB, Phenomenex Strata C18-E, and Agilent ABS Elut-NEXUS), and a liquid/liquid extraction with acetonitrile. A reverse-phase HPLC with a C18 column and gradient elution program was used. MS detection was carried out in MRM mode. Quantitation was performed using the internal standard method. Validation parameters were very satisfactory. The high selectivity and sensitivity of the method (LOQ < 9.5 ng mL−1, and LOD < 3.3 ng mL−1) allowed us to determine target analytes even in small amounts. Precision, matrix effect, and stability were also evaluated. The whole validated method has finally been applied to the analysis of 194 real samples of human amniotic fluid obtained from pregnant women (15–21 weeks of gestation) in order to monitor the effective intake of target analytes: 96% of the examined women consumed caffeine during pregnancy while a lower percentage (20%) showed acetaminophen intake. The whole procedure is simple and easy to perform with minimal sample preparation and short analysis time.

Simultaneous Determination of 11 Illicit Phenethylamines in Hair by LC-MS-MS: In Vivo Application.

Existing phenethylamines are a class of synthetic compounds that differ from each other only in small changes to a largely conserved chemical structure. The recreational and illicit use of phenethylamines is a widespread problem. A simple procedure for the simultaneous quantitative determination in hair of 11 phenethylamines that are officially recognized as illicit by Italian legislation (p-methoxyamphetamine; p-methoxymethamphetamine; 3,4,5-trimethoxyamphetamine; 2,5-dimethoxyamphetamine; 2,5-dimethoxy-4-methylamphetamine; 2,5-dimethoxy-4-ethylamphetamine; 2,5-dimethoxy-4-bromoamphetamine; 2,5-dimethoxy-4-bromophenethylamine; 2,5-dimethoxy-4-iodophenethylamine; 2,5-dimethoxy-4-ethylthiophenethylamine and 2,5-dimethoxy-4-n-propylthiophenethylamine) has been developed and validated. Extraction from the matrix was performed after incubation in methanolic HCl and filtered reconstituted extracts were injected into a liquid chromatography/tandem mass spectrometry system (LC-MS-MS) without any further purification steps. This validated LC-MS-MS method has been used to determine the in vivo accumulation/retention of the above target analytes in hair after repeat oral administration to rats. This experiment further permitted investigation of the effect of pigmentation on the uptake of these phenethylamines by hair and the effect of hair pigmentation. The developed method could potentially be used for forensic and toxicological purposes, in the detection and quantitation of these illicit substances in human hair in workplace drug testing; drug-facilitated crime investigation; driver re-licensing; determining drug abuse history and postmortem toxicology.