Recovery and Stability of Δ9-Tetrahydrocannabinol Using the Oral-Eze® Oral Fluid Collection System and Intercept® Oral Specimen Collection Device.

Oral fluid (OF) is increasingly used for clinical, forensic and workplace drug testing as an alternative to urine. Uncertainties surrounding OF collection device performance, drug stability and testing reproducibility may be partially responsible for delays in the implementation of OF testing in regulated drug testing programs. Stability of Δ(9)-tetrahydrocannabinol (THC) fortified and authentic specimens was examined after routine collection, transport and laboratory testing. Acceptable recovery and stability were observed when THC-fortified OF (1.5 and 4.5 ng/mL) was applied to Oral-Eze devices. Neat OF samples collected with Oral-Eze, processed per the package insert, and fortified with THC (3 and 6 ng/mL) were stable (±20%) at room temperature (21-25°C), refrigerated (2-8°C) and frozen (-25 to -15°C) conditions up to 1 month, while samples collected with Intercept devices showed decreases at refrigerated and room temperatures. After long-term refrigerated or frozen storage, maximum reductions in THC concentrations were 42% for Oral-Eze and 69% for Intercept. After ≥1 year frozen storage, 80.7% of laboratory specimens positive for THC (3 ng/mL cut-off) by GC-MS were reconfirmed positive (within 25%), with an average THC decrease of 4.2%. Specimens (n = 47) processed with Oral-Eze (diluted) and tested via enzyme immunoassay were concordant with LC-MS-MS results and showed 100% sensitivity and 95% specificity. Paired specimens collected with Oral-Eze and Intercept exhibited 98% overall agreement between the immunoassay test systems. Collectively, these data demonstrate consistent and reproducible recovery and stability of THC in OF after collection, transport and laboratory testing using the Oral-Eze OF Collection System.

Concentrations of Morphine and Codeine in Paired Oral Fluid and Urine Specimens Following Ingestion of a Poppy Seed Roll and Raw Poppy Seeds.

Interpretation of opiate drug test results can be challenging due to casual dietary consumption of poppy seeds, which may contain variable opiate content. Opiate concentrations in paired oral fluid (OF), collected with the Oral-Eze(®) Oral Fluid Collection System, and urine were analyzed after ingestion of poppy seeds from the same source, consumed raw or contained in a roll. In Part 1, 12 individuals consumed equal portions of a poppy seed roll. For Part 2, the same individuals consumed an equivalent quantity of raw poppy seeds, containing ∼3.2 mg of morphine and 0.6 mg of codeine. Specimens were analyzed both by enzyme immunoassay (opiates) and by GC-MS (morphine/codeine). Urinary morphine was between 155-1,408 (roll) and 294-4,213 ng/mL (raw), measured at 2, 4, 6 and 20 h post-ingestion. Urinary codeine concentrations between 140-194 (roll) and 121-664 ng/mL (raw) were observed up to 6 h post-ingestion. Following consumption of raw poppy seeds, OF specimens were positive, above LOQ, from 0.25 to 3.0 h with morphine ranging from 7 to 600 ng/mL and codeine from 8 to 112 ng/mL. After poppy seed roll consumption, morphine concentrations of 7-143 ng/mL were observed up to 1.5 h with codeine detected in only 5.5% of OF specimens and ranging from 8 to 28 ng/mL. Combined with the existing poppy seed literature, these results support previous findings and provide guidance for interpretation of OF opiate testing.

Psychiatric effects and psychoactive substance use in anabolic-androgenic steroid users.

The purpose of this study was to assess the psychiatric effects of anabolic-androgenic steroid (AAS) use and assess the frequency of other psychoactive substance use in a population of AAS users compared with non-AAS-using weight-lifter controls. One hundred sixty-four subjects were administered a demographic survey, including psychiatric history, substance use history, AAS use history, and medical history. Psychiatric diagnoses were made and psychological testing was performed. User categories were determined by history and urine testing. The user categories did not differ significantly on psychological testing. Past AAS users had a higher incidence of psychiatric diagnosis than the nonuser and current user groups. Hypomania was correlated with AAS use, and major depression with AAS discontinuation. Present psychoactive substance abuse or dependence was relatively low across all user categories. AAS dependence was seen in 12.9% of current users and 15.2% of past users of AAS. In conclusion, AAS use may lead to psychiatric disorders in certain individuals. Concurrent use of psychoactive drugs other than AAS does not appear to be common in intensively training weight lifters and bodybuilders.

Human liver cocaine esterases: ethanol-mediated formation of ethylcocaine.

A new, pharmacologically active metabolite of cocaine, ethylcocaine, has been reported in individuals after concurrent use of cocaine and ethanol. Formation of ethylcocaine may contribute to the common coabuse of these two drugs and the apparent danger of this practice. We have identified a nonspecific carboxyl-esterase that catalyzes the ethyl transesterification of cocaine to ethylcocaine in the presence of ethanol. In the absence of ethanol, this human liver esterase catalyzes the hydrolysis of cocaine to benzoylecgonine, a metabolite that is inactive as a psychomotor stimulant. A second human liver esterase is also described. This enzyme catalyzes hydrolysis of cocaine to ecgonine methyl ester, also inactive as a stimulant. These two liver esterases may play important roles in regulating the metabolic inactivation of cocaine.

Functional tolerance to the hypothermic effect of phenobarbital in rats.

Phenobarbital elicits a hypothermic effect in rats. To determine if functional tolerance develops to this effect on temperature, rats were treated twice daily with IP injections of sodium phenobarbital (PheB) or saline. The PheB doses were increased over 21 days and then were held constant for another 23 days. On the next day (day 45) animals from both groups were given 80, 110 or 160 mg/kg PheB IP and the decrease in rectal temperature after 2 h was determined. Animals were decapitated after the temperature measurement and brain PheB levels were determined. A 1.51-fold shift in the relationship between brain level and response was found for the group given chronic PheB. These results show functional tolerance occurs to the hypothermic effect of PheB. This experiment was done in animals that were on a restricted food regimen. Rats given chronic PheB lost more weight than the group given chronic saline unless extra food was provided. We found that this occurred because the rats given chronic PheB lost more food through the wire cage floor than rats given chronic saline.

Sudden cardiac death during exercise in a weight lifter using anabolic androgenic steroids: pathological and toxicological findings.

A 21-year-old, previously healthy weight lifter collapsed during a bench press workout. He had taken anabolic androgenic steroids parenterally for the previous several months. Pertinent autopsy findings included marked cardiac and renal hypertrophy and hepatosplenomegaly, with regional myocardial fibrosis and focal myocardial necrosis. Nandrolone (19-nor-testosterone) metabolites were identified in postmortem urine. The possible etiologies of the cardiac findings are discussed.

Localization of sites in periventricular forebrain mediating cardiovascular effects of gamma-aminobutyric acid agonists and antagonists in anesthetized cats.

Previous studies have shown that injection of gamma-aminobutyric acid (GABA) antagonists such as bicuculline methiodide (BMI) into the forebrain (i.e., lateral and third) ventricular system elicits neurally mediated increases in blood pressure and heart rate and inhibits baroreflex bradycardia in anesthetized cats. Similar administration of muscimol, a GABA agonist, blocks or reverses the effects of BMI but has no effect on blood pressure or heart rate in untreated animals. These findings suggest that GABAergic inhibition may tonically suppress a forebrain mechanism capable of modifying autonomic outflow to the cardiovascular system. In the present study, we used a technique designed to restrict the distribution of intraventricularly administered drugs to varying degrees in order to better localize the relevant sites of drug action. Our findings show that BMI increases heart rate and blood pressure and that muscimol counters these changes by acting at a site that is accessible from the intermediate (as opposed to the rostral or caudal) region of the third ventricle. In contrast, these agents influence baroreflex bradycardia by acting at more rostral periventricular sites. These findings are consistent with the notion that the GABAergic mechanisms involved in the cardiovascular effects resulting from intraventricular administration of BMI and muscimol are located in the periventricular hypothalamus.

Microinjection of GABA antagonists into posterior hypothalamus elevates heart rate in anesthetized rats.

Microinjection of the GABA antagonist bicuculline methiodide 1-25 ng into the posterior hypothalamus of urethane-anesthetized rats evoked sympathetically-mediated increases in heart rate of up to 150 beats/min and modest increases in blood pressure which could be prevented by prior local microinjection of muscimol 50 ng. Microinjection of picrotoxin but not strychnine produced similar effects. These results suggest that a latent sympathoexcitatory mechanism in this region is tonically inhibited by endogenous GABA.

A simplified, colorimetric micromethod for xylose in serum or urine, with phloroglucinol.

We have developed a simplified xylose assay procedure that requires only 10 min and requires 50 microL of serum or 5 microL of urine. The reaction with phloroglucinol is more sensitive than the classic p-bromaniline color reaction, and requires only 4 min of heating for color development. A single reagent is mixed with the specimen directly, without prior protein precipitation. Analytical recovery of xylose added to serum was quantitative; precision studies resulted in a between-day coefficient of variation of 5.2%. Glucose, which has significant potential for interference in most other xylose procedures, reacts under the test conditions only to the extent of 70 mumol of apparent xylose per liter for a 5.5 mmol/L solution of glucose. The new procedure has been valuable in the assessment of malabsorption, especially in children and infants, where serum xylose is the preferred measurement.

High-pressure liquid chromatographic assay of chloramphenicol in biological fluids.

Chloramphenicol can be assayed by a sensitive and selective high-pressure liquid chromatographic assay technique. The method described is easily adapted to analysis of many other drugs.