Epithelial cell mitotic arrest--a useful postmortem histologic marker in cases of possible colchicine toxicity

Following ingestion of 30 mg of presumed benztropine (Cogentin) a 39-year-old male developed nausea, vomiting and diarrhea. His admission to hospital was soon followed by collapse and death. Histological examination, however, revealed increased numbers of mitotic figures in otherwise normal epithelial cells of the esophagus and bronchioles, a feature characteristic of colchicine toxicity. Subsequent toxicological analyses confirmed the presence of colchicine in the urine, but not in the blood. A dispensing error had resulted in substitution of colchicine for Cogentin. Histological findings had, therefore, provided evidence of colchicine toxicity and had guided subsequent toxicological evaluation. In suspected cases of colchicine toxicity, histological samples should, therefore, be taken from multiple sites along the gastrointestinal and respiratory tract in addition to other organs and tissues so that diagnostic morphological changes can be looked for.     

Serum cannabinoid levels 24 to 48 hours after cannabis smoking

Low concentrations of THC and 11-hydroxy-THC in serum samples are often claimed not to result from recent cannabis use. Prediction of time of exposure is difficult, especially if distinctive features of drug use could not be observed. Therefore, the aim of the study was to investigate the presence of THC and 11-hydroxy-THC in serum samples as well as to obtain preliminary data on the analyte profile for a time window of 24-48 hours after discontinuation of cannabis smoking. Serum samples from heavy (n = 12, > 1 joint/day), moderate (n = 11, < or = 1 joint/day) and light (n = 6, < 1 joint/week) smokers of cannabis were analyzed for THC, 11-hydroxy-THC and free THC-COOH by GC/MS as well as for glucuronidated THC-COOH by LC/MS-MS. The blood samples were collected 24-48 hours after abstaining from cannabis use. Additionally, 8 specimens were obtained from persons after discontinuation of the drug for more than 48 hours. During collection of the blood samples, distinctive effects due to drug use could not be observed. For heavy users of cannabis, THC was detectable in 8 samples, and in 5 cases both biologically active compounds, THC and 11-hydroxy-THC, were present (1.3-6.4 ng THC/mL serum, 0.5-2.4 ng 11-hydroxy-THC/mL serum). Among moderate users, in 1 sample 1.8 ng THC/mL serum and 1.3 ng 11-hydroxy-THC/mL serum were determined, and another sample was tested positive with low concentrations close to the limit of detection. In serum samples of light users both analytes could not be detected, indicating that in those persons a positive finding of THC and 11-hydroxy-THC may rather result from recent consumption than from cannabis use 1 or 2 days prior to blood sampling. The concentrations of THC-COOH and its glucuronide covered a wide range in all groups of cannabis users. However, there was a trend to higher concentrations in heavy users compared to moderate users, and the mean concentration was smaller in light smokers than in moderate smokers. Overall, the findings indicated that data from pharmacokinetic studies should be supplemented by data obtained from "real-life" samples.     

Use of Xylazine in Drug‐Facilitated Crimes

Human xylazine poisoning is uncommon. This report describes the use of xylazine for intentional poisoning with criminal intent. Two incidents occurred within 3 weeks: the first involved one victim, and the second involved two victims. The clinical presentations were brief coma, bradycardia, hypotension, and hyperglycemia. The victims recalled having been given a drink from a stranger in a hospital waiting room before loss of consciousness. In the first case, general drug screening by gas chromatography/mass spectrometry (MS) revealed xylazine in the gastric contents, but liquid chromatography–tandem MS (LC‐MS/MS) of serum did not. In the second incident, LC‐MS/MS screening of both victims’ urine and serum samples revealed an unknown peak in the total ion chromatograms, which a molecular mass database identified as morantel or xylazine. The latter was confirmed by comparison with a xylazine standard. Based on this report, we suggest that xylazine should be classified as a controlled drug.     

Godden v Hales revisited – James II and the dispensing power

This article considers whether the King's Bench decision in Godden v Hales (1686) allowing James II to dispense with the application of the Test Acts was correct. Most modern historians believe that the decision was either correct, or at least a justifiable interpretation of the legal precedents; contemporaries overwhelmingly believed the decision was wholly wrong. This article explains the nature and importance of the dispensing power, and considers the relevant case law precedents in detail. It concludes that the line of authorities forbidding the king from dispensing with the anti-simony statutes should have been applied to the Test Acts. The accusations of illegal and unconstitutional conduct relating to the dispensing power made against James II during the 1688 Revolution and in the Declaration of Rights were thus wholly justified.     

Effect of murine retroviral infection on hair and serum levels of cocaine and morphine.

LP-BM5 retrovirally infected female C57BL/6J mice were administered cocaine, morphine or both by daily intraperitoneal injection for 9 weeks. Drug concentrations were measured by radioimmunoassay in serum and in hair extracts. Hair samples obtained from all drug-treated mice were positive for the drug injected, while none of the saline-treated mice had detectable drug levels in hair or serum. The average morphine concentration obtained from non-infected mice was 11 ng/mg hair whereas the amount found in the LP-BM5-infected mice was significantly higher (20 ng/mg hair). Mice injected with both morphine and cocaine were given 50% of the regular dose of each drug and drug levels in the hair of these animals were approximately half that of mice injected with the full dose of the single drug. Non-infected mice treated with both drugs had a mean value of 7 ng morphine/mg hair and 374 ng cocaine/mg hair while retrovirus-infected mice had significantly higher concentrations, 10 ng morphine/mg hair and 1160 ng cocaine/mg hair (P less than 0.001). Serum concentrations of cocaine and morphine were significantly higher (P less than 0.01) in the retrovirus-infected animals from 40 min to 1.5 h. The increased concentrations of cocaine and morphine in serum during retrovirus infection are accompanied by a significant increase in the amount of drug incorporated into the hair matrix. This change indicates that retroviral infection may influence the disposition of these drugs in the systemic circulation.     

Olanzapine concentrations in clinical serum and postmortem blood specimens--when does therapeutic become toxic?

The concentration of olanzapine (Zyprexa) was determined in 1653 clinical serum specimens during routine drug monitoring, and in 58 postmortem whole blood specimens as part of routine toxicological analysis. The analysis of olanzapine was performed by the solid-phase extraction of 1.0 mL of buffered serum or blood, followed by gas chromatography separation with nitrogen-phosphorus detection. The analysis of the clinical serum samples showed that 86% of positive serum values were within the range of 5 to 75 ng/mL, with a mean and median of 36 and 26 ng/mL, respectively. These data suggest that the concentrations of olanzapine expected during therapy may be higher than those previously reported. In 58 postmortem whole blood specimens the mean olanzapine concentration was 358 ng/mL with a range of 10 to 5200 ng/mL. Further, investigation of deaths involving olanzapine suggest that potential toxicity should be considered at concentrations above 100 ng/mL. Although the majority of the olanzapine-related deaths were associated with many other drugs, death primarily due to olanzapine toxicity was determined at concentrations in post-mortem blood as low as 160 ng/mL.     

The detection of 6-monoacetylmorphine in urine, serum and hair by GC/MS and RIA

6-Monoacetylmorphine (6-MAM) is a good indicator for the intake of heroin and can be detected in blood, urine and hair of heroin users. A new radioimmunoassay (RIA) designed specifically for 6-monoacetylmorphine (6-MAM) was tested for its usefulness for the quantitation of the drug in urine, serum and hair. Its cross-reactivity with heroin and its metabolites, and related compounds was also determined. Eighty-nine hair, six serum and 25 urine samples where 6-MAM had been previously identified by GC/MS were analysed for 6-MAM with the new RIA kit. A good correlation existed between the GC/MS and RIA results for the hair samples. However, the amount of 6-MAM found in serum and urine differed considerably between the two methods. This difference could be explained by the cross-reactivity of the antibody with morphine and morphine-6-glucuronide, which are present in much larger amounts in serum and urine, than in hair. To evaluate a new rationalisation procedure, some hair samples were split into two portions after incubation. One part was analyzed for 6-MAM by RIA, and the other portion by GC/MS.     

Comparative investigations on the uptake of basic substances into the human keratinocyte cell line HaCaT--a model for the uptake of xenobiotics into the keratinocytes of the hair follicle

The human cell line HaCaT was used to study drug uptake by keratinocytes as an in-vitro model to elucidate drug incorporation into anagen hair follicle. The basic drugs under investigation were taken up very rapidly resulting in a concentration plateau in the keratinocytes which was dependent on the drug concentration in the cell culture medium. The results obtained for HaCaT clearly demonstrated the existence of a partition-equilibrium between the extracellular and intracellular drug concentrations. Only small amounts of the offered drugs were taken up and remarkable differences were observed showing a decreasing uptake for imipramine > haloperidol > cocaine/benzoylecgonine. Total protein content in the culture medium was 3.5 +/- 0.3 mg/mL and the protein binding of the drugs to the foetal serum proteins was found to be negligible in the experiments. Overall, the in vitro findings were consistent with previous observations for in vivo drug incorporation into hair. In particular, an explanation was found for the correlation between the AUC-value and hair concentration observed in animal studies as well as for the generally low drug concentrations in non-pigmented hair.     

Correction to Tzidony and Ravreby (1992): A Statistical Approach to Drug Sampling: A Case Study

Abstract:  In 1992, Tzidony and Ravreby presented a confidence interval for the total weight of a seizure of illicit drugs present in a population. Their approach has subsequently been applied by several researchers in the field. The formula on which their approach is based does, however, not fully take into account the proportion of drug units found in the sample. In this paper, a modification is presented that consistently uses the correct sample size in all terms of the confidence interval, based on the proportion of drug units found in the sample. The effective sample size is smaller than the original sample size, and this should consequently be accounted for in the estimation of the standard error and in the corresponding t -distribution. The new confidence interval is again based on the assumption that the proportion of drug units in the population is known after sampling.     

Liquid/solid extraction on diatomaceous earth for drug analysis in postmortem blood

Absorption extraction on diatomaceous earth was examined and found to be compatible with typical postmortem blood specimens encountered in forensic toxicology. The effects of solvent, solvent volume, eluate flow rate, and pH on drug recovery and extract quality were investigated. It was concluded that the best method was not necessarily that with the highest recoveries, but that considerations of extract quality were also required. The optimized method was compared with a single-step liquid/liquid extraction method and found to be superior in terms of ease of operation, extract quality, and absolute recovery. The results indicated also that although useful for screening, quantitative methods using liquid/solid extraction may be prone to error.     

Serum beta-glucuronidase activity in polytrauma patients and in centrifuged autopsy blood samples]

The serum activity of beta-glucuronidase was investigated in 58 patients after severe trauma as well as in 43 autopsy cases. In 10 cases the enzyme activities in postmortem blood samples from the femoral vein were compared to those present in the correspondent heart blood samples. An elevated activity of beta-glucuronidase was observed in 14% of the patients within the first 36 h after severe trauma increasing to 62% in blood samples collected later on. The activity of beta-glucuronidase in the heart blood samples was always higher than in the corresponding sample from the femoral vein. In cases of prolonged post-mortem interval an elevated activity might have been due to bacterial contamination. In postmortal blood samples from the femoral vein an elevated enzyme activity was found in 70% of the study material. The results of the preliminary study on the activity of beta-glucuronidase in blood samples frequent in forensic routine work indicated that an elevated enzyme activity might be present for the following scenery: after severe trauma, in alcohol/drug abuse, presence of putridity/autolysis, presence of inflammatory processes, in diabetes as well as in carcinoma diseases. The significance of elevated beta-glucuronidase activity concerning alterations of unconjugated drug concentration due to in vitro cleavage of O-glucuronides should be investigated.     

药品安全的刑法保护

在当前药品安全的严峻形势下，刑法作为保障药品安全的最后一道防线，应当进一步强化对药品安全的保护作用。尽管《刑法修正案（八）》对药品安全的本罪进行了相应的修改，但现行刑法关于药品安全本罪的规定依然存在犯罪归属体系错误、主观罪过范围过窄、犯罪行为规定不全面、罚金刑规定过于原则、资格刑缺失等不容忽视的问题，针对这些问题刑法还应当作出进一步的修正，以期实现对药品安全的周全保护。     

Estimation of gamma-hydroxybutyrate (GHB) co-consumption in serum samples of drivers positive for amphetamine or ecstasy

There is no toxicological analysis of gamma-hydroxybutyrate (GHB) applied routinely in cases of driving under influence (DUI); therefore the extent of consumption of this drug might be underestimated. Its consumption is described as occurring often concurrently with amphetamine or ecstasy. This study examines 196 serum samples which were collected by police during road side testing for GHB. The samples subject to this study have already been found to be positive for amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and/or 3,4-methylenedioxyethamphetamine (MDEA). Analysis has been performed by LC/MS/MS in the multiple reaction monitoring (MRM) mode. Due to its polarity, chromatographic separation of GHB was achieved by a HILIC column. To differentiate endogenous and exogenous levels of GHB, a cut-off concentration of 4μg/ml was applied. Of the 196 samples, two have been found to be positive for GHB. Of these samples, one sample was also positive for amphetamine and one for MDMA. Whilst other amphetamine derivates were not detected in these samples, both samples were found to be positive for cannabinoids. These results suggest that co-consumption of GHB with amphetamine or ecstasy is relatively low (1%) for the collective of this study.     

Toxicologic findings in suicide with doxepin and paroxetine]

A young nurse was found dead in her flat. In chemical-toxicological analysis the following femoral blood drug concentrations were determined: paroxetine 0.176 mg/l, doxepine 82.12 mg/l, desmethyldoxepine 0.34 mg/l. Additionally the drug concentrations were determined in various body fluids and organs. The results of the described fatality are discussed. For interpretation of toxicologic results in antidepressant fatalities ratios of parent drug to metabolite and postmortem drug redistribution should be taken into account.     

Detection of psilocin in body fluids

Active compounds of some mushrooms e.g. Psilocybe cubensis, Paneolus subalteatus or Stropharia coronilla, the psychotropic agents psilocybin and psilocin, have hallucinogenic effects. In one case of 'magic mushroom' intake, we had to analyse blood and urine. Psilocin was detected in the urine with REMEDi HS. Most of the psilocin was excreted as the glucuronide. Therefore an enzymatic hydrolysis should be the first step in analysis. Free psilocin was determined at a concentration of 0.23 mg/l while the total amount was 1.76 mg/l urine. The concentration of psilocin in serum was too low for detection with REMEDi HS. We proved a GC-MS-method with d(3)-morphine as internal standard and silylation with MSTFA. Similarly to urine, most of the psilocin in serum was found in the conjugated form. The concentration of free psilocin was 0.018 mg/l, that of total psilocin, 0.052 mg/l serum.     

Analyses of Beverage Remains in Drug Rape Cases Revealing Drug Residues ‐ The Possibility of Contamination from Drug Concentrated Oral Fluid or Oral Cavity Contained Tablets

In drug‐facilitated sexual assault (DFSA) cases, drug residues may be detected in beverage remains found in cups or glasses known to have been used by the victims. In this small naturalistic study, the possibility of beverages being contaminated, either by drug concentrated oral fluid or by oral cavity contained tablets, was investigated. Analysis of residues from cups containing soft drinks was performed by immunoassay and ultra‐performance liquid chromatography‐mass spectrometry (UPLC‐MS/MS). Beverage with both added tablets and spiked oral fluid was investigated, as well as simulation of swallowing tablets. Only the residues from added tablets were positive with immunoassay, while drugs were detectable in all cups using more sensitive UPLC‐MS/MS. In conclusion, the possibility of detecting drug residues in beverages due to a contamination, from either drug concentrated oral fluid or oral cavity contained tablets at a time of consumption, should be kept in mind when performing sensitive analysis.     

Minimizing analytical interferences from digoxin-like immunoreactive substances (DLIS) in cases of digoxin toxicity

Recently, the value of therapeutic drug monitoring for digoxin has been called into question by the finding of endogenous digoxin-like immunoreactive substances (DLIS) in the serum of individuals, especially premature and full-term neonates, not being treated with digoxin. In some cases, values have been as high as 10 micrograms/L. Levels as high as 20 micrograms/L and 80 micrograms/g can be found in bile and meconium. Because of the magnitude of this interference, it is essential that methods be developed for measuring digoxin in the presence of DLIS. This is particularly important when such analyses are required in forensic science cases of suspected digoxin toxicity. This report outlines the high performance liquid chromatographic (HPLC) and radioimmunoassay (RIA) methods that we used in assessing the relative contribution made by digoxin, its metabolites, and DLIS to serum and tissue digoxin concentrations obtained by RIA in a forensic pediatric case of suspected digoxin toxicity.     

A micromethod for the isolation of drugs from blood using amberlite XAD-2

The extraction of drugs from small blood samples (1 ml or less) for subsequent quantitative determination is described. Isolation was carried out by adsorption of the drugs to Amberlite XAD-2 resin utilizing a batch procedure that enabled the simultaneous extraction of up to 200 samples in approx. 5 hours. A new desorption technique yielded extracts of high purity that could be used directly for gas chromatographic or radioimmunological determinations, even if hemolyzed or putrid blood was to be examined. The following 26 substances were quantitated after addition to postmortem blood speciments at concentrations of 1-10 microgram/ml: tilidine, diphenhydramine, dibenzepine, imipramine, chlorpromazine, amphetamine, pentazocine, phenacetin, methaqualone, meprobamate, parathion, diazepam, digoxin, beta-methyldigoxin, carbromal, glutethimide, amobarbital, pentobarbital, cyclobarbital, phenobarbital, diphenylhydantoin, carbutamide, tolbutamide, glycodiazin, tolazamide and chlorpropamide. Thereby recoveries of 60-100% could be achieved. The reproducibility of the procedure was satisfactory as demonstrated by coefficients of variation of 3.7-8%.