Was a child poisoned by ethanol? Discrimination between ante-mortem consumption and post-mortem formation.

The presence of ethanol in human specimens collected during autopsies is generally considered as an indication of recent ante-mortem alcohol consumption. The interpretation of the results may however be impaired by post-mortem formation of ethanol when microorganisms capable of fermentation of glucose to ethanol are present. Since the distribution in the different fluids and tissues remains contentious to conclude on the origin of the detected ethanol, the determination of specific metabolites of ethanol such as ethyl glucuronide (EtG) may be performed to discriminate between exogenous (ante-mortem) and endogenous (post-mortem). Toxicological analysis of specimens from the autopsy of a child aged 14 months displayed a high concentration of ethanol in blood and tissues. In order to discriminate between ante-mortem alcohol administration and post-mortem formation, the presence of microorganisms capable of ethanol production was checked by fermentation tests and the liver was tested for the presence of EtG and compared with a positive control. Fermentation tests displayed in the blood of the deceased the presence of the bacterial strain Lactococcus garvieae capable of producing ethanol from glucose. The absence of EtG in the liver of the deceased compared to the high level (19.56 mug/g) detected in the positive control's liver is a further indication that the ethanol detected in the body of the deceased is of post-mortem origin.

Formation of scopolamine from N-butyl-scopolammonium bromide in cigarettes.

Scopolamine (hyoscine) is a naturally occurring alkaloid found in solonacea, the so-called "night shade" plants. Therapeutic applications of scopolamine are in ophthalmology to cause mydriasis and for the prevention of motion sickness, among others. It is known to induce hallucinogenic effects at a high dose. The N-butyl bromide derivative of scopolamine, available commercially as Buscopan, is commonly used as an antispasmotic. The possibility of forming scopolamine from N-butyl-scopolammonium bromide when burning cigarettes fortified with Buscopan was investigated based on a record of a prison inmate who claimed to experience hallucinations after smoking Buscopan. Liquid chromatography-tandem mass spectrometry in electrospray ionization mode was used to monitor the formation of scopolamine. Various series of eight cigarettes spiked with 10 mg of N-butyl-scopolammonium bromide with and without filters and in different smoking modes were investigated. The smoke of the burning cigarettes, the ashes, and the filter were analyzed for the presence of scopolamine. Scopolamine was detected in all cases.

Ethyl glucuronide concentration in hair is not influenced by pigmentation.

This work shows that the concentration of ethyl glucuronide (EtG) in hair, a marker for the evaluation of the alcohol consumption, is not influenced by the presence or absence of melanin. The results confirm that, unlike many other substances, the EtG determination in hair has not to take into account the hair colour for the correct interpretation of hair testing results.

Determination of nevirapine and efavirenz in plasma using GC/MS in selected ion monitoring mode.

An accurate, selective, and sensitive method for the determination of the nonnucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine (nvp) and efavirenz (efv) in human plasma using gas chromatography-mass spectroscopy in selected ion monitoring mode (GC/MS-SIM) was developed. Solid-phase extraction (SPE) gave extraction yields near 100% for both nvp and efv, and calibration curves were linear over the therapeutic concentration ranges. Variation of intraday and interday precision was below 5%. Intraday and interday inaccuracies varied between 0.6% and 10.4%. The lower limits of detection using a 200-microL plasma sample were 27 ng/mL for nvp and 26 ng/mL for efv, and the lower limits of quantification were 54 ng/mL and 72 ng/mL, respectively. The method was applied to the determination of nvp and efv in plasma specimens of 73 patients in HIV stage III or IV and on antiretroviral treatment in Kigali, Rwanda.

HPLC analysis of generic antiretroviral drugs purchased in Rwanda.

A reversed phase HPLC method using photo diode array detection for the simultaneous quantification of lamivudine, stavudine, nevirapine, zidovudine, methyl paraben and propyl paraben in solid and liquid drug formulations was developed and validated. The separation was achieved using a Waters Symmetry C8 column, using a mobile phase gradient comprising 50 mM NaH2PO4 (pH 3.8) and acetonitrile (95:5 to 45:55, v/v) and a flow gradient (0.5 to 1.0 ml/min). The limits of detection and quantification were below 19 ng/ml and 55 ng/ml respectively. The intra- and inter-day assay precisions were within 4.4% relative standard deviations. The developed method was applied to 12 different generic antiretroviral medications, consisting of tablets, capsules and solutions, produced by two Indian manufacturers and purchased by the Central Agency of Essential Drug Procurement of Rwanda for the ESTHER project in Rwanda. The average content of the antiretroviral agent(s) compared to the labeled amount(s) was 101.4%. Methyl paraben and propyl paraben, added to solutions as preservatives, were within the FDA recommended limits.