Effect of cimetidine on verapamil disposition.

The effects of multiple doses of cimetidine on single-dose verapamil kinetics were studied in nine healthy men. Baseline hepatic blood flow was estimated by indocyanine green elimination on day 1. On day 2, the subjects received verapamil, 10 mg iv, after which the plasma concentration-time profile was determined. After a 2-day washout, cimetidine, 300 mg, was taken by mouth four times a day for 5 days. The indocyanine green study was repeated on day 9 and verapamil was taken on day 10. Cimetidine reduced verapamil clearance by 21% and increased the elimination t1/2 by 50%. The volume of distribution at steady state did not change. Cimetidine increased hepatic blood flow in some subjects, while decreasing it in others. There was no correlation between individual changes in verapamil clearance and hepatic blood flow. These data indicate that cimetidine reduces verapamil clearance by mechanism(s) other than a change in hepatic blood flow or volume of distribution.

The effects of a single dose of amphetamine and iprindole on the serotonergic system of the rat brain.

Rats treated with iprindole (IPR) (10 mg/kg, i.p.) were given a single dose (15 mg/kg, i.p.) of amphetamine (AMP). Marked decreases in the activity of tryptophan hydroxylase (TPH) were observed in both the cerebral cortex and neostriatum after 6 hr, with maximum depression observed at 24 hr. Enzyme activity had returned to control levels in neostriatum after 3 days and in cerebral cortex after 7 days. Levels of serotonin (5-HT) in both the cerebral cortex and neostriatum were significantly lowered at 24 hr but had recovered by 72 hr. Levels of tryptophan (TRP) in the cortex were significantly elevated after 6 hr, recovering by 24 hr. In the neostriatum, the activity of tyrosine hydroxylase (TH) was significantly depressed by 24 hr and remained so for 7 days. Concentrations of dopamine (DA) were decreased at all times examined. This study clarifies the differences previously observed in the response of the serotonergic system to amphetamine or methamphetamine (METH) in iprindole-treated rats.

Studies on the mechanism of tolerance to methamphetamine.

We have reported that the ability of high doses of methamphetamine to impair dopamine and serotonin synthesis in the rat brain is attenuated when animals are pretreated with gradually increasing doses of methamphetamine. To examine the mechanism of this tolerance phenomenon, the effect of methamphetamine on several neurochemical parameters was determined in naive and methamphetamine-pretreated rats. The elevation of nigral substance P concentrations by methamphetamine was attenuated in pretreated compared to naive rats. The methamphetamine-induced reduction in [3H]sulpiride binding in the rat neostriatum and nucleus accumbens was similarly attenuated in animals pretreated with methamphetamine. Determination of brain concentrations of methamphetamine and amphetamine revealed significantly lower concentrations of both compounds in the brains of pretreated compared to naive animals. The results indicate a reduction in the ability of methamphetamine to increase dopamine transmission in the brains of methamphetamine-pretreated rats. Furthermore, this effect appears to be due, at least in part, to a change in the disposition of methamphetamine in pretreated animals.

The acute effects of methamphetamine, amphetamine and p-chloroamphetamine on the cortical serotonergic system of the rat brain: evidence for differences in the effects of methamphetamine and amphetamine.

Cortical tryptophan hydroxylase (TPH) activity was reduced 3 h after a 10 or 15 mg/kg i.p. dose of either amphetamine (AMP), methamphetamine (METH), or p-chloroamphetamine (PCA). These injections of METH or PCA also decreased cortical serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) concentrations; none of the four doses of AMP decreased indoleamine concentrations. The time course of the effects following a 15 mg/kg dose of each amine was also different. Cortical TPH activity was reduced by all three amines for periods up to 24 h, whereas only METH and PCA significantly decreased 5-HT and 5-HIAA concentrations for long periods. These data suggest that each of the amphetamines may inhibit TPH activity, whereas only METH and PCA produced long-lasting decreases in indoleamine concentrations, reflecting either varying degrees of toxicity or differential effects of AMP on enzyme activity and neurotransmitter concentrations.

The effects of phencyclidine on glutamic acid decarboxylase activity in several regions of the rat brain.

Glutamic acid decarboxylase (GAD) activity in several regions of the rat brain were monitored after administration of phencyclidine. Sub-acute (4 injections over 12 h) treatment decreased cerebellar GAD activity 6 and 12 h after the last dose; recovery was noticed by 24 h. This effect occurred with doses of 5 and 10 mg/kg. GAD activity in other brain regions was not affected by this treatment. Acute and chronic treatments with phencyclidine caused no change in GAD activity in any of the brain regions examined.

Advances in forensic toxicology.

Over the last 15 years there have been many changes in the practice of forensic toxicology. One of the most noteworthy has been the recognition of the need for good laboratory practices in the forensic toxicology laboratory. This has resulted in the development of an accreditation program for laboratories. Increasingly, forensic toxicologists are asked to interpret results in driving under the influence of drug cases. These interpretations are also difficult because of the lack of data correlating blood (or plasma) concentrations with impairment. The development of newer immunoassays and hyphenated mass spectrometric techniques now allow the forensic toxicologist to assay a large number of drugs (both traditional and products of the biotechnology revolution) with increasing sensitivity. This article focuses on these changes and some of the challenges facing the forensic toxicologists of the 21st century.

High-pressure liquid chromatographic determination of chloridazepoxide and its major metabolites in biological fluids.

A simple, isocratic, reversed-phase, high-pressure liquid chromatographic procedure was developed for the determination of chlordiazepoxide and its major metabolites in plasma and urine. The within-run coefficient of variation was 3.4-8.0%, and the day-to-day variation was 4.0-8.0%. Recoveries of 80-91% with sensitivity limits of 50 ng/ml were obtained for the parent drug and its metabolites. Plasma and urine samples collected after single intravenous and single oral doses were analyzed using this procedure.

Drugs in fatally injured young male drivers.

One or more drugs were detected in 81 percent of 440 male drivers, aged 15-34, killed in motor vehicle crashes in California; two or more drugs were detected in 43 percent. Alcohol, the most frequently found drug, was detected in 70 percent of the drivers, marijuana in 37 percent, and cocaine in 11 percent. Each of 24 other drugs was detected in fewer than 5 percent. Except for alcohol, drugs were infrequently found alone; typically, they were found in combination with high blood alcohol concentrations. The causal role of drugs in crashes was assessed by comparing drivers with and without drugs in terms of their responsibility for the crash. Alcohol was associated with increased crash responsibility; the role of other drugs could not be adequately determined.

Determination of methaqualone and its major metabolite in plasma and saliva after single oral doses.

A gas liquid chromatographic procedure for the determination of methaqualone and metabolite l (2-methyl-3(2-hydroxymethylphenyl)-4-(4H)-quinazoline) in plasma and saliva is described. The method requires 1.0 mL of sample and involves internal standard addition, extraction with butyl chloride, chromatography on a 1% SP-1,000 column packing using nitrogen phosphorous detectors. Within-run and day-to-day coefficient variations were less than 8% and the procedure has a sensitivity limit of 20 ng/mL. The method has been used to monitor both parent drug and metabolite in plasma and samples collected after a single oral dose.

An HPLC method for the quantitation of quinidine and its metabolites in plasma: an application to a quinidine-phenytoin drug interaction study.

A rapid and simple method is described for the separation of quinidine (Q), its metabolites 3-hydroxyquinidine (30HQ) and 2'oxoquinidinone (QD) as well as the quinidine salt contaminant, dihydroquinidine (DHQ) in plasma by high pressure liquid chromatography (HPLC). A silica column was used with methanol: 1 N ammonium nitrate: 2 N ammonium hydroxide (28:1:1 v/v) as the mobile phase to separate the compounds and a fluorescence spectrophotometer for detection. Application of this method is demonstrated by the analysis of human plasma samples collected from a volunteer during a phenytoin-quinidine drug interaction study.

Gas chromatography-chemical ionization mass spectrometry of cocaine and its metabolites in biological fluids.

A gas chromatographic-chemical ionization mass spectrometric (GC-CIMS) method is described for the determination of cocaine, benzoylecgonine, and norcocaine. The procedure uses stable isotopes as internal standards and a mixture of methane-ammonia as chemical ionization reagent gas. Run-to-run and within-run coefficients of variation (%) are less than 10% and the method has a sensitivity of less than 5 ng/mL from 1 mL or 1 gram of sample. The procedure has been applied to a number of cases involving cocaine intoxication and analytical data from these are described.

Quantitative analysis of emetine and cephaeline by reversed-phase high performance liquid chromatography with fluorescence detection.

A versatile method for the quantitation of emetine and cephaeline in biological samples is described. Two milliliters of samples containing N-propylprocainamide as the internal standard are buffered to pH 9 and extracted with n-butyl chloride. After subsequent back extraction into 0.01 M hydrochloric acid, a portion of the acid layer is analyzed by reversed-phase high performance liquid chromatography with fluorescence detection. Routinely, the minimum level of detection for both drugs is 5 ng/mL and linearity is demonstrated from 5 to 2500 ng/mL.

Stability of delta 9-tetrahydrocannabinol (THC), 11-hydroxy-THC, and 11-nor-9-carboxy-THC in blood and plasma.

The stabilities of delta 9-tetrahydrocannabinol (THC) and two of its metabolites, 11-hydroxy-delta 9-tetrahydrocannabinol (HO-THC) and 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (COOH-THC), were determined in blood and plasma stored at -10 degrees C, 4 degrees C, and room temperature. Each of the cannabinoids was added to freshly-drawn blood and plasma to give concentrations of 20 ng/mL. Two-mL aliquots were stored in silanized tubes and the cannabinoid concentrations were monitored by gas chromatography/mass spectrometry over a 6-month period. No significant changes were observed in the concentrations of the cannabinoids for the first month of storage. However, the concentrations of THC and HO-THC in blood stored at room temperature had decreased significantly at 2 months. No statistically significant changes were detected in cannabinoid concentrations in plasma or blood stored at 4 degrees or -10 degrees C for up to 4 months. After 6 months at room temperature, the blood concentrations of THC and HO-THC had decreased by 90 and 44%, respectively, whereas the concentration of COOH-THC was not significantly different from the control. The possibility of loss of cannabinoids from blood due to adsorption onto the grey stoppers used in Venoject tubes was also investigated. Over a 24-hr period, no significant differences were detected in any of the cannabinoid concentrations regardless of sample size (1.3 or 8 mL), differences in temperature (-10 degrees C, 4 degrees C, or room temperature), or extent of contact with the tube's stoppers.

The screening and quantitation of diazepam, flurazepam, chloridazepoxide, and their metabolites in blood and plasma by electron-capture gas chromatography and high pressure liquid chromatography.

A combined GLC-ECD and HPLC procedure has been developed for the analysis of the most commonly encountered benzodiazepine drugs and has been applied to both plasma and postmortem blood samples. There is no doubt that since their introduction the use of these sensitive analytical methods have resulted in an increase in the incidence of detection of these drugs in both clinical and forensic toxicology cases.

Profiles of delta 9-tetrahydrocannabinol metabolites in urine of marijuana users: preliminary observations by high performance liquid chromatography-radioimmunoassay.

Metabolic profiles of 11-nor-9-carboxylic acid-delta 9-tetrahydrocannabinol (COOH-THC) and other THC metabolites were determined in an infrequent and a frequent marijuana user by high performance liquid chromatography-radioimmunoassay (HPLC-RIA). In the infrequent user, no unconjugated COOH-THC was detected in urine samples for the first 8 h following smoking, whereas this metabolite was detected in the urine samples from a frequent user. A metabolite was also detected in the frequent user, which was not present in the urine sample from the infrequent user.

Proficiency testing in forensic toxicology: a feasibility study.

This study has shown that a national proficiency testing program in forensic toxicology is feasible. Samples that resemble typical case specimens were prepared and shipped to approximately 100 laboratories. Participation varied between 61 and 73%. Tissue samples obtained from laboratory animals can be used to simulate those encountered by forensic toxicologists. This has been demonstrated using liver homogenates from animals administered pentobarbital and methaqualone and propoxyphene and acetaminophen. There was a large coefficient of variation however, for the quantitation of acetaminophen in liver. The qualitative data obtained during the course of this study showed a very low incidence of false positives. However, there was a disappointingly low percentage of positive responses for (a) low concentrations of secobarbital and (b) the opiate narcotics (morphine and codeine) in blood, despite the fact that sensitive immunoassay procedures are available for detecting these particular compounds in blood samples. The quantitative determination of drugs and metabolites, other than ethanol, shows wide interlaboratory variation. This variation is presumably not a result of the use of different analytical techniques, since gas liquid chromatography was used by the majority of participants to quantitate drugs and metabolites. Forensic toxicologists are willing to participate in a voluntary proficiency testing program conducted by an independent agency. The performance data developed in this study can serve as a baseline for current forensic toxicology laboratory functional capability in the assessment of future changes and improvements in analytical forensic toxicology.

High performance liquid chromatography-immunoassay of delta 9-tetrahydrocannabinol and its metabolites in urine.

High performance liquid chromatographic-immunoassay (HPLC-IA) profiles of cannabinoid metabolites in urine samples were obtained using four different antisera. The urines were chromatographed on a reverse phase system using a gradient of acetonitrile in water (pH 3.3) and fractions collected every 30 s. Some urine samples were hydrolyzed with methanolic sodium hydroxide before fractionation. Peaks of immunoreactivity were detected at a fraction corresponding to 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (COOH-THC) and at an early eluting fraction; however, the profiles depended upon the specificity of the antisera used.

Concentrations of lidocaine and monoethylglycylxylidide (MEGX) in lidocaine associated deaths.

Concentrations of lidocaine and MEGX were determined in a variety of tissues and other samples collected at autopsy. In 13 of the cases examined in which lidocaine was associated with death, tissue concentrations were greater than 15 mg/kg. Tissue concentrations in other patients treated with lidocaine were significantly lower.

Drugs and driving: a systematic analytical approach.

To collect useful epidemiological data about drug involvement in highway safety, it is essential that sensitive and specific analytical procedures be used to establish the presence of and to determine the concentrations of drugs and metabolites in samples collected from drivers. This paper describes a comprehensive and systematic screening procedure requiring 6 mL of blood, which has been used for the analysis of samples collected from injured and fatally injured drivers. The procedure uses radioimmunoassay, gas chromatography with selective detectors, and high performance liquid chromatography. Drugs and metabolites presumptively identified are then confirmed primarily using gas chromatography--chemical ionization mass spectrometry.

Toxicological findings in a fatal overdose of verapamil.

Presented is a case where the death was attributed to the deliberate ingestion of an overdose of verapamil (V). Blood, urine, and gastric concentrations of the drug were determined by gas chromatography with nitrogen phosphorus detection (GC-NPD). Identification of norverapamil (NV) was made. A presumptive identification of o-demethylnorverapamil (DNV) was also made.