Metodi di screening, conferma e separazione enantiomerica di nuovi designer drugs amfetamino-simili

Amphetamine designer drugs are central nervous system stimulants widely diffused worldwide in illegal markets. Their identification is often difficult, because of the high structural variety of new compounds. Monitoring of drugs of abuse in biological fluids is successfully used for clinical and forensic applications. In this work a method for identify and quantify 3,4-methylenedioxy-N-isopropylamphetamine (MDIP), 3,4-methylenedioxy-N-cyclopropylmethylamphetamine (MDCPM), 3,4-methylenedioxy-N-benzylamphetamine (MDBZ) and 3,4-methylenedioxy-N-ethylamphetamine (MDE) in urine, and a method for identify and quantify 4-bromo-2,5,beta-trimethoxyphenethylamine (BOB), 4-methyl-2,5,beta-trimethoxyphenethylamine (BOD), 3,4-methylenedioxy-beta-methoxyphenethylamine (BOH) and 4-methyl-2,5-dimethoxy-beta-hydroxyphenethylamine (BOHD) in plasma are described. In addition, the cross-reactivity profiles of 41 new amphetamines to 6 commonly used screening tests for urine and oral fluid are reported. Furthermore an easy-to-prepare chiral CE method for the enantiomeric separation of 13 new amphetamine-like designer drugs using cyclodextrins as chiral selectors has been developed.</br

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 mother's 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 child's 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 &lt; 9.5 ng mL−1, and LOD &lt; 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

Identification and quantitation of 4-bromo-2,5-dimethoxyamphetamine in seized blotters

Blotters are usually impregnated with hallucinogens such as lysergic acid diethylamide (LSD); only rarely other psychoactive substances are detected. In this work we identified 4-bromo-2,5-dimethoxyamphetamine (DOB) and 2,5-dimethoxyamphetamine (DMA) in illicit blotters seized in Italy. This report describes a rapid method for the simultaneous identification and quantitation of DOB and its precursor (DMA) by liquid chromatography tandem mass spectrometry (LC–MS–MS), using 2,3-dimethoxyphenethylamine-d3 as internal standard. Regression equations were linear over the tested concentration range with good correlation coefficients. The achieved levels of sensitivity may be suitable to confirm the possible presence of DOB and DMA also in low concentration or in traces in seized material for forensic analysis. The developed method showed good reproducibility and sensitivity, and could be used for similar routine analysis. To our knowledge, this is the first report describing the detection of DOB and DMA from blotters

Comparative Evaluation of the Autonomic Response to Cognitive and Sensory Stimulations through Wearable Sensors

Psychological stress is known to activate the autonomic nervous system (ANS), thus representing a useful target to be monitored to understand the physiological, unconscious effect of stress on the human body. However, little is known about how differently the ANS responds to cognitive and sensory stimulations in healthy subjects. To this extent, we enrolled 23 subjects and administered a stress protocol consisting of the administration of sensory (olfactory) and cognitive (mathematical) stressors. Autonomic parameters were unobtrusively monitored through wearable sensors for capturing electrocardiogram and skin conductance signals. The results obtained demonstrated an increase of the heart rate during both stress protocols, with a similar decrease of the heart rate variability. Cognitive stress test appears to affect the autonomic parameters to a greater extent, confirming its effects on the human body. However, olfactory stimulation could be useful to study stress in specific experimental settings when the administration of complex cognitive testing is not feasible

Effects of homeopathic mother tinctures on breath alcohol testing

In some countries, it is illegal to drive with any detectable amount of alcohol in blood; in others, the legal limit is 0.5 g/L or lower. Recently, some defendants charged with driving under the influence of alcohol and have claimed that positive breath alcohol test results were due to the ingestion of homeopathic mother tinctures. These preparations are obtained by maceration, digestion, infusion, or decoction of herbal material in hydroalcoholic solvent. A series of tests were conducted to evaluate the alcoholic content of three homeopathic mother tinctures and their ability to produce inaccurate breath alcohol results. Nine of 30 subjects gave positive results (0.11–0.82 g/L) when tests were taken within 1 min after drinking mother tincture. All tests taken at least 15 min after the mother tincture consumption and resulted in alcohol-free readings. An observation period of 15–20 min prior to breath alcohol testing eliminates the possibility of false-positive results

Enantiomeric separation of 13 new amphetamine-like designer drugs by capillary electrophoresis, using modified: cyclodextrins

An easy-to-prepare chiral CE method for the enantiomeric separation of 13 new amphetamine-like designer drugs, using CDs as chiral selectors, was developed. Sulfated-β-CD was found to be the best chiral selector among the three used (sulfated-β-CD, caroboxymethyl-β-CD, dimethyl-β-CD). The separation of the analytes was achieved in a fused-silica gel capillary at 20 °C using an applied voltage of +25 kV. The optimized background electrolyte consisted of 63.5 mM H3PO4 and 46.9 mM NaOH in water. Several electrophoretic parameters such as CD type, CD concentration (1 - 40 mg/mL), buffer pH (2.6, 3.6, 5.0, 6.0), length of the capillary (70 - 40 cm total length), amount of the organic solvent (methanol and acetonitrile) were investigated and optimized

Screening method for five commonly used amphetamines in urine by NMR spectroscopy

In the recent years the so-called designer drugs have been diffused dramatically worldwide. Amphetamine psychostimulants are one of the major classes of illicit drugs consumed for recreational purposes in the world. The detection of the illegal substances is important both in the management of their misuse and in the forensic field. Urine matrix is a reliable biological matrix for verifying amphetamines intake in the short and medium term because the excretion of these drugs mainly occurs in urine, where substantial amounts of unchanged drug are present. In response to a growing demand for reliable evidence of amphetamine use, a method for identification and quantitation of amphetamine, methamphetamine, 3,4-methylenedioxy-N-methylamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxy-N-ethylamphetamine in urine has been developed by nuclear magnetic resonance spectroscopy, using electronic reference to access in vivo concentration method. The method showed good linearity. The limits of detection and quantitation of amphetamines analyzed ranging from 1.9–7.9 to 5.8–23.8 &#181;g mL−1, respectively. These data demonstrate that the present method is suitable for detection of these drugs in urine samples, allowing to quantitate the target analytes without any purification step of the matrix

An LC&#8212;MS&#8212;MS method for quantitation of four new phenethylamines (BOX series) in plasma: in vivo application

The appearance of new “designer drugs” in the illicit market poses a serious health risk because they have unknown safety profiles, have a high potential for abuse, high potency, and can lead to devastating health consequences. For this reason, it is desirable to develop validated and reliable analytical screening tests that allow detection of amphetamines and related designer drugs in biological samples. We report a method for separation and quantitation of four new phenethylamines, 4-bromo-2,5-beta-trimethoxyphenethylamine (BOB), 4-methyl-2,5-beta-trimethoxyphenethylamine (BOD), 3,4-methylenedioxy-beta-methoxyphenethylamine (BOH), and 4-methyl-2,5-dimethoxy-beta-hydroxyphenethylamine (BOHD), in plasma. Quantitation was achieved via liquid chromatography&#8212;tandem mass spectrometry (LC&#8212;MS&#8212;MS) in the multiple reaction monitoring mode, using 2,3-dimethoxyphenethylamine-d 3 as internal standard. The method was validated according to international guidelines. The parameters determined were selectivity, sensitivity, matrix effect, linearity, precision, recovery, and stability. All parameters were satisfactory. To remove matrix interference, solid-phase extraction was introduced in the method as clean-up step. The same method was applied in a pharmacokinetic study to monitor the target compounds in rat plasma after a single oral administration. The developed and validated LC&#8212;MS&#8212;MS method is the first available for quantitation of BOB, BOH, BOD, and BOHD in a biological matrix. This method is recommended for use in forensic and clinical toxicology, because of its sensitivity, selectivity, and simplicity. An important extension of this method could involve its application to other complex matrices

Magnetic resonance imaging of the brain and skull of sheep with cerebral coenurosis

Objective: To determine MRI characteristics of the skulls and brains of sheep with chronic cerebral coenurosis (CC) caused by naturally acquired Taenia multiceps infection. Animals: 33 sheep with CC and 10 healthy control sheep. Procedures: Sheep underwent MRI of the head. Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were determined. For CC-affected sheep, the number, location, and volume of T. multiceps cysts were determined and the percentage volumes of cysts in the cranial cavity and rostral and caudal fossas of the cranial cavity were calculated. Focal and diffuse abnormalities of cranial bones in CC-affected sheep were identified. Brain edema and hemorrhage and signs of increased cranial pressure (ICP) in MRI images were determined. Results: Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were significantly larger for CC-affected sheep versus healthy control sheep. Total volumes of cysts ranged from 4.40% to 46.93% in cranial cavities of sheep, 4.12% to 51.53% in rostral fossas of cranial cavities of sheep, and 15.24% to 68.30% in caudal fossas of cranial cavities of sheep. Moderate to severe diffuse cranial bone abnormalities and signs of increased ICP in MRI images were detected in 21 and 24 sheep, respectively, and were positively correlated with cyst volumes. Conclusions and Clinical Relevance: Results suggested that cranial cavity volume and morphological abnormalities can be detected in sheep with CC. These changes may reflect abnormalities in ossification of the cranial bones secondary to chronically increased ICP caused by development of T. multiceps cysts