Increased DNA Methylation of ABCB1, CYP2D6, and OPRM1 Genes in Newborn Infants of Methadone-Maintained Opioid-Dependent Mothers.

Objective: To investigate whether in utero opioid exposure, which has been linked to adverse neurodevelopmental and social outcomes, is associated with altered DNA methylation of opioid-related genes at birth.

Study Design: Observational cohort study of 21 healthy methadone-maintained opioid-dependent mother-infant dyads consecutively delivered at >36 weeks of gestation, and 2 comparator groups: smoking, "deprived" opioid-naïve mother-infant dyads (n = 17) and nonsmoking, "affluent" opioid-naïve mother-infant dyads (n = 15). DNA methylation of ABCB1, CYP2D6, and OPRM1 genes for mothers and babies was determined from buccal swabs.

Variations in Infant CYP2B6 Genotype Associated with the Need for Pharmacological Treatment for Neonatal Abstinence Syndrome in Infants of Methadone-Maintained Opioid-Dependent Mothers.

 Neonatal abstinence syndrome (NAS) in infants of methadone-maintained opioid-dependent (MMOD) mothers cannot be predicted in individual cases. We investigated whether variation in infant genotype is associated with severity of NAS.  This is a pilot observational cohort study of 21 MMOD mothers and their newborns.

Effects of oral fluid contamination on two oral fluid testing systems.

Oral fluid is a popular matrix for drug testing; however, little has been published concerning the effect that food or beverages may exert on oral fluid screening tests. This study describes the effects of 19 different foods, beverages and vinegars on two test systems, the Concateno Certus and Orasure Intercept. Samples giving positive screening results were subjected to confirmatory analysis using gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry.

Distribution of opiates in femoral blood and vitreous humour in heroin/morphine-related deaths.

The distribution of free morphine (FM), codeine and 6-acetylmorphine (6AM) in vitreous humour (VH) and femoral blood (FB) was measured in 70 cases involving heroin/morphine. The relationship between tissue drug concentrations was assessed with respect to case circumstances. Total morphine (TM) concentrations in FB are also reported.

Analysis of skeletal muscle has potential value in the assessment of cocaine-related deaths.

This study assesses the interpretive value of cocaine, benzoylecgonine (BZE) and cocaethylene (COET) in skeletal muscle (rectus femoris) in cocaine-using decedents. The distribution of these analytes in cardiac muscle (CM), vitreous humour (VH), femoral blood (FB) and cardiac blood (CB) is also reported. In rectus femoris muscle, the spatial distribution of the analytes was examined across the whole rectus femoris muscle collected from seven fatalities in which cocaine was detected.

Hollow-fiber liquid-phase microextraction and gas chromatography-mass spectrometry of barbiturates in whole blood samples.

Here, we present a method for measuring barbiturates (butalbital, secobarbital, pentobarbital, and phenobarbital) in whole blood samples. To accomplish these measurements, analytes were extracted by means of hollow-fiber liquid-phase microextraction in the three-phase mode. Hollow-fiber pores were filled with decanol, and a solution of sodium hydroxide (pH 13) was introduced into the lumen of the fiber (acceptor phase).

Determination of opiates in whole blood and vitreous humor: a study of the matrix effect and an experimental design to optimize conditions for the enzymatic hydrolysis of glucuronides.

Undoubtedly, whole blood and vitreous humor have been biological samples of great importance in forensic toxicology. The determination of opiates and their metabolites has been essential for better interpretation of toxicological findings. This report describes the application of experimental design and response surface methodology to optimize conditions for enzymatic hydrolysis of morphine-3-glucuronide and morphine-6-glucuronide.

Validation of a gas chromatography-ion trap-tandem mass spectrometry assay for the simultaneous quantification of cocaine, benzoylecgonine, cocaethylene, morphine, codeine, and 6-acetylmorphine in aqueous solution, blood, and skeletal muscle tissue.

A gas chromatography-ion trap-tandem mass spectrometry method was developed and validated for the simultaneous solid-phase extraction and quantification of cocaine, benzoylecgonine, cocaethylene, morphine, codeine, and 6-monoacteylmorphine in blood, muscle tissue, and water. An assay for total morphine in blood and muscle was also validated. The limit of quantification was ≤ 0.

The effect of sodium fluoride preservative and storage temperature on the stability of 6-acetylmorphine in horse blood, sheep vitreous and deer muscle.

This study examined the in vitro stability of 6-acetylmorphine (6AM) in horse blood, sheep vitreous humour (VH) and homogenised deer muscle stored under different storage conditions. The stability of 6AM in horse blood is of interest because many toxicological laboratories utilise this matrix for the preparation of blood calibration and check standards and the latter are typically stored during routine use. Data on the storage stability of 6AM in human VH is extremely limited and no data has been reported in muscle.

The effect of sodium fluoride preservative and storage temperature on the stability of cocaine in horse blood, sheep vitreous and deer muscle.

The in vitro stability of cocaine in horse blood, sheep vitreous humour (VH) and homogenised deer muscle is described. The stability of cocaine in horse blood was of interest because many toxicology laboratories utilise horse blood for the preparation of calibration and check standards and the latter are typically stored during routine use. The storage stability of cocaine in human VH and muscle has not been previously reported.

CYP2B6 and OPRM1 gene variations predict methadone-related deaths.

The largest proportion of methadone-associated deaths occurs during the drug induction phase. We analysed methadone-related fatalities for gene variations linked with methadone action. A significant association between high methadone concentrations and the CYP2B6*6 allele characteristic of the slow metabolizer phenotype was identified.

Detection of ketamine and its metabolites in urine by ultra high pressure liquid chromatography-tandem mass spectrometry.

Current analytical methods used for screening drugs and their metabolites in biological samples from victims of drug-facilitated sexual assault (DFSA) or other vulnerable groups can lack sufficient sensitivity. The application of liquid chromatography, employing small particle sizes, with tandem mass spectrometry (MS/MS) is likely to offer the sensitivity required for detecting candidate drugs and/or their metabolites in urine, as demonstrated here for ketamine. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) was performed following extraction of urine (4 mL) using mixed-mode (cation and C8) solid-phase cartridges.

Retention reproducibility of basic drugs in high-performance liquid chromatography on a silica column with a methanol-high-pH buffer eluent. Changes in selectivity with the age of the stationary phase.

The long-term reproducibility of the separation of basic drugs by high-performance liquid chromatography using a silica column with a methanol-high-pH eluent was examined. The relative capacity factors of specific tertiary amines changed systematically compared with the other drugs on storage of the columns for 1 year. The changes were enhanced on storing the column in mobile phase and were accelerated by prolonged washing with water.

The effect of salbutamol on breath alcohol testing in asthmatics.

Subjects suffering from asthma can occasionally experience difficulty in providing adequate breath samples for evidential breath alcohol testing devices and may therefore resort to the use of bronchodilators such as salbutamol to improve their respiration. Experiments showed that although salbutamol caused bronchodilation it did not affect breath alcohol levels of asthmatics who have been drinking. The blood:breath alcohol ratios obtained from asthmatics were within the normally recorded range before and after use of salbutamol.

Study into the ability of patients with impaired lung function to use breath alcohol testing devices.

A subject who fails to provide an adequate breath sample for a breath alcohol measuring device under the provisions of Road Traffic Act 1988 may be charged with refusing to supply a sample unless the police officer believes the person is genuinely unable to do so. Subjects who are confronted with this situation may approach their general practitioner or chest physician for advice on whether they are medically able to provide adequate breath samples to satisfy the breath testing devices. There is currently no guidance available for medical practitioners concerning respiratory performance or lung function which will impair the use of such breath testing devices.

The response of evidential breath alcohol testing instruments with subjects exposed to organic solvents and gases. III. White spirit exposure during domestic painting.

The concentration of white spirit vapour in the breathing zone of a volunteer engaged in domestic painting activities has been investigated. Air was continuously sampled and analysed using an infra-red gas analyser (Miran) during painting experiments. Both indoor and outdoor painting tasks were undertaken using commercial paints in a wide variety of situations.

The response of evidential breath alcohol testing instruments with subjects exposed to organic solvents and gases. II. White spirit and nonane.

Following exposure to white spirit vapour, the effect of the expired solvent on evidential breath alcohol equipment was investigated under controlled exposure chamber conditions and in a simulated painting exercise. Five volunteers inhaled the solvent in an exposure chamber at a concentration of 100ppm for periods up to 4h 17min. Two other volunteers were exposed to white spirit while painting with domestic gloss paint in an unventilated room under which conditions exposure concentrations reached 185ppm for 20min.

The response of evidential breath alcohol testing instruments with subjects exposed to organic solvents and gases. I. Toluene, 1,1,1-trichloroethane and butane.

Experimental work has been undertaken to investigate the potential interference of toluene, 1,1,1-trichloroethane and butane with the evidential breath alcohol testing instruments used in Great Britain (Lion Intoximeter 3000 and Camic Breath Analyser). Volunteers inhaled the volatile substances in an exposure chamber for up to 4 hours, at concentrations of 100, 350 and 600ppm respectively. Subsequently breath was tested on leaving the chamber.

Whole blood quality assurance control samples for forensic toxicology.

Pilot studies were undertaken to examine the feasibility of preparing freeze-dried whole blood samples containing drugs of interest to be used as assay controls. The availability of such samples will enable analysts and laboratory managers to monitor analytical performance. A pilot batch of freeze-dried blood containing five compounds of forensic significance and a blank blood sample were prepared for reconstitution to 2.

The effect of respiratory aerosol inhalers and nasal sprays on breath alcohol testing devices used in Great Britain.

Subjects suffering from respiratory diseases occasionally experience difficulty when attempting to provide breath samples to satisfy the requirements of breath alcohol testing devices. In order to assist ventilation, such subjects may resort to the use of an aerosol inhaler. Twenty aerosol inhalers containing drugs which exhibit an effect on the respiratory system and five nasal sprays used to relieve the symptoms of hay fever, have been tested for interference on the range of breath alcohol measuring devices used by the police in Great Britain.

Comparison of methods for the analysis of tricyclic antidepressants in small whole blood samples.

A variety of liquid-liquid and solid-phase extraction techniques have been compared for the extraction of tricyclic antidepressant drugs from small whole blood samples. Each of the procedures was evaluated with respect to extract cleanliness, extraction efficiency, and reproducibility. The three most efficient methods were compared on a variety of authentic whole blood case samples containing drugs at therapeutic and toxic levels.