Intoxication caused by new psychostimulants: analytical methods to disclose acute and chronic use of benzofurans and ethylphenidate

International Journal of Legal Medicine - The acute and chronic toxicity of several new psychoactive substances (NPS) is unknown, and only little information is available on the pharmacology and...     

Assessment of prenatal exposure to ethanol by meconium analysis: results of an Italian multicenter study

Background: This study estimated in 7 Italian cities the prevalence of prenatal exposure to ethanol by determining fatty acid ethyl esters (FAEEs; palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, and arachidonic esters) and ethyl glucuronide (EtG) in neonatal meconium samples. Methods: A total of 607 meconium samples were obtained from neonatal wards of 7 public hospitals: Verona and San Daniele del Friuli in the northeast of the country, Reggio Emilia in the middle east, Florence and Rome in the center, and Naples and Crotone in the southwest of the peninsula. Meconium biomarkers were assessed by a validated methodology using liquid chromatography–tandem mass spectrometry and the results categorized using the accepted cutoff of 2 nmol/g total amount of 7 FAEEs and 2 nmol/g EtG, to differentiate between heavy maternal ethanol use during pregnancy and occasional or no use at all. Results: On the basis of the above‐reported cutoffs, the overall prevalence of newborns prenatally exposed to maternal ethanol was 7.9%: 0% in Verona, 4.0% in San Daniele del Friuli, 4.9% in Naples, 5.0% in Florence, 6.2% in Crotone, up to 10.6% in Reggio Emilia, and 29.4% in Rome. Low maternal education level and younger maternal age were associated with biomarker scores over the cutoff. There was also a significant correlation between the highest percentage of prenatal exposure in the capital and certain maternal sociodemographic characteristics. Conclusions: These results indicate considerable variability in the prevalence of fetal exposure to ethanol in different Italian cities, as determined by the objective measurement of biomarkers in meconium. These data, together with previous ones obtained in Barcelona, Spain, indicate that gestational ethanol exposure is widespread, at least in parts of Europe.     

Assay of gamma-glutamylcysteine synthetase activity in Plasmodium berghei by liquid chromatography with electrochemical detection

This work describes a high-performance liquid chromatography (HPLC) method to determine γ-glutamylcysteine (γ-GC), the intermediate product of glutathione biosynthesis. Separation relies on isocratic reversed-phase chromatography using a Symmetry C18 HPLC column, particle size 5 μm, 4.6×250 mm i.d. The mobile phase is methanol–dibasic sodium phosphate (pH 6.6; 2.8 mM) (10:90, v/v) at the flow-rate of 0.5 ml/min and detection is operated electrochemically (+200 and +550 mV) with a pre-column derivatisation reaction using ortho-phthalaldehyde (OPA) as reagent. Under these conditions the calibration range of γ-GC was 0.3–10 μg/ml; the limit of quantification was 0.3 μg/ml; accuracy, expressed as %Bias, was <10 and precision (%CV) was <6. The proposed HPLC assay was used to quantitate the γ-glutamylcysteine produced by the γ-glutamylcysteine synthetase of the rodent malaria parasite Plasmodium berghei in an in vitro enzymatic assay.     

Liquid chromatography–electrospray ionization mass spectrometry determination of methylphenidate and ritalinic acid in conventional and non-conventional biological matrices

A procedure based on liquid chromatography–electrospray ionization mass spectrometry is described for determination of methylphenidate (MPH) and its principal metabolite ritalinic acid (RA) in plasma, urine, oral fluid and sweat using 3,4-methylendioxypropylamphetamine (MDPA) as internal standard. Aliquots of 100 μL biological fluids and sweat patch were initially treated with acetonitrile, centrifuged, and clear supernatants evaporated and redissolved in 10 mM ammonium acetate. Chromatography was performed on a reversed-phase column using a gradient of 10 mM ammonium acetate and acetonitrile as a mobile phase at a flow rate of 1 mL/min. Separated analytes were confirmed and quantified by positive electrospray ionization mass spectrometry and selected ion monitoring acquisition mode. Limits of quantifications were 1 ng/mL plasma, 1 ng/sweat patch, 0.5 ng/mL oral fluid and urine for MHF; 1 ng/mL plasma and oral fluid, 1 ng/sweat patch, 0.5 ng/mL urine for RA using 100 μL biological fluids or one sweat-patch per assay. Calibration curves were linear over the calibration ranges for both MPH and RA, with r² > 0.99. At three concentrations spanning the linear dynamic range of the assay, mean recoveries ranged between 67.9–90.3% for MPH and 36.3–92.4% for RA in the different biological matrices. This method was applied to therapeutic monitoring of MHP and RA in conventional and non-conventional biological matrices from individuals in drug treatment.     

Neonatal withdrawal syndrome after chronic maternal drinking of mate

The premature newborn of a mother who reported drinking mate during pregnancy presented with increased jitteriness and irritability, high-pitched cry, hypertonia in the limbs, and brisk tendon reflexes consistent with neonatal withdrawal syndrome. High concentrations of caffeine and theobromine were detected in various maternal and neonatal biological matrices (placenta, cord serum, neonatal urine, maternal and neonatal hair, meconium, and breast milk), demonstrating both acute and chronic prenatal and postnatal exposure to these methylxanthines, contained in high amounts in homemade mate. Symptoms progressively disappeared at 84 hours of age, although intermittent irritability was still present when the infant was discharged at 24 days of age. Fluctuating caffeine (and theobromine) content in different breast milk feeds likely generated the baby's irritability, due to either the physiological stimulatory effects of the methylxanthines or postnatal withdrawal syndrome as the substances cleared from the body. The mother was strongly advised to initiate a considerable, progressive, constant reduction of mate consumption to a maximum of 2 cups a day for the duration of breastfeeding.     

Systemic poisoning caused by percutaneous absorption of mineral spirit]

Owing to a car accident, the clothes of a injured joiner were permeated with a timber impregnating product containing 51.8% of mineral spirit (a mixture of naphthenes, aromatic and aliphatic hydrocarbons). Despite a short-lasting skin exposure (approximately 40 minutes), dermal contact has caused full thickness burns that, in their turn, have made easier the percutaneous absorption and the storage of organic solvents in subcutaneous tissue depots. Twenty-four hours later, clinical findings of neurologic involvement have arisen, that have got worse when the peripheral tissue perfusion has got better by the adequate replacement of lost blood.