A validated method for the analysis of cannabinoids in post-mortem blood using liquid-liquid extraction and two-dimensional gas chromatography-mass spectrometry

Phenazepam is a long-acting benzodiazepine that, unlike other benzodiazepines, is currently not scheduled as a narcotic in Finland, most other European countries or the USA. It is used as an anxiolytic, sedative-hypnotic and anti-epileptic, mainly in Russia. In Finland, as well as in some other countries, an increase in the unauthorized use of phenazepam has been observed in recent years. In the one year period between July 1, 2010 and June 30, 2011 the prevalence of phenazepam in Finland was assessed among drivers apprehended for driving under the influence of drugs (DUID), in medico-legal autopsy cases and in police confiscations of illicit drugs. In DUID cases an LC-MS/MS method preceded by solid phase extraction was used for the determination of phenazepam. In the post-mortem investigations the sample preparation consisted of liquid-liquid extraction followed by derivatization and the determination was carried out by GC-MS. The police confiscations were analysed by GC-MS. There were 141 positive phenazepam cases among apprehended drivers, representing approximately 3.5% of all confirmed drug cases (n=4007) in this time period. The median (range) phenazepam blood concentration in DUID cases was 0.061 mg/L (0.004-3.600 mg/L). The median phenazepam concentration in cases with no concomitant stimulant use was significantly higher than the overall median concentration. Phenazepam was found in 17 medico-legal autopsy cases and the median (range) blood concentration was 0.048 mg/L (0.007-1.600 mg/L). Phenazepam was not considered by the medico-legal team to be the sole cause of death in any of the cases, the majority of them being accidental opiod overdoses. There were 26 seizures of phenazepam by the Police in the time period studied, some of the batches consisted of a mixture of phenazepam and stimulant designer drugs. The data show that phenazepam abuse is a widespread phenomenon in Finland. A typical user was a male multi-drug user in his 30s. The concentration range of phenazepam among apprehended drivers and medico-legal autopsy cases was wide and the drug was usually found along with other psychoactive drugs. Therefore, although it seems likely that phenazepam contributed to impairment of driving in some DUID cases, the extent of its effect remains unclear and further studies are needed to define the concentrations causing impairment and toxicity.     

A simple identification method of saliva by detecting Streptococcus salivarius using loop-mediated isothermal amplification

We previously reported that detection of Streptococcus salivarius is feasible for proving the presence of saliva in a forensic sample. Here, a simple and rapid method for the detection of S. salivarius in forensic samples was developed that uses loop-mediated isothermal amplification (LAMP). The LAMP primer set was designed using S. salivarius-specific sequences of glucosyltransferase K. To simplify the procedure, the sample was prepared by boiling and mutanolysin treatment only, and the entire analytical process was completed within 2.5 h. The cut-off value was set at 0.1 absorbance units, measured at 660 nm, upon termination of the reaction. S. salivarius was identified in all saliva samples, but was not detected in other body fluids or on the skin surface. Using this method, S. salivarius was successfully detected in various mock forensic samples. We therefore suggest that this approach is useful for the identification of saliva in forensic practice.     

Chemical toxicological analysis of urinary cannabinoids by chromatography mass spectrometry

A chromato-mass-spectrometric method for identification of cannabinoids and 9-carboxy-11-nor-delta 9-tetrahydrocannabinol by their methyl derivatives is proposed. Use of alkaline hydrolysis in pretreatment of samples ensures the optimal conditions for complete isolation of 9-carboxy-11-nor-delta 9-tetrahydrocannabinol. Derivation with methyl iodide in absolute acetone in the presence of potassium carbonate was used. Preliminary conclusion on the presence of cannabinoids in the urine can be made from the results of enzyme immunoassay.     

The analysis of clingfilms by infrared spectroscopy and thermal desorption capillary gas chromatography.

An automated thermal desorption gas chromatography technique has been adapted to analyse traces of volatile compounds in proprietary food-wrapping films. Fourteen brands of polyvinylchloride film, seven brands of polyethylene film and one polyvinylidene chloride film were discriminated. Prior infrared analysis was used to identify the polymer type. The chromatograms showed minor changes in volatiles along the length of a roll of film and major changes in films exposed to daylight or in contact with cannabis resin.     

A simple histochemical technique for the identification of gunshot residue

Alizarin red S (ARS) is a commonly used organic dye useful in the histologic identification of calcium deposits. ARS also forms colored reaction products with other metal ions, including barium and lead, which are present in primer residue. In histochemical studies, ARS is shown to identify primer residues from several manufacturers as well as primer residue deposited in tissue, either experimentally or in close-range gunshot wounds. This can be easily accomplished with routine histologic techniques. ARS does not stain gunpowder residue, tattoo pigment, melanin, graphite, india ink, or anthracotic pigment.     

A quantitative differentiation method for acrylic fibers by infrared spectroscopy

Absorbance peak areas of nitrile (2240 cm(-1)), carbonyl (1730 cm(-1)) and CH (1370 cm(-1)) groups were obtained for 48 colorless acrylic fibers by infrared (IR) microspectroscopy. The carbonyl signal, related to the comonomers most commonly used in acrylic fibers, was ratioed against the nitrile and CH bands, pertaining to the backbone of the polymer chains. The ratios A1730/A2240 and A1730/A1370, a relative measure of the comonomer content in the fiber, were used to differentiate the samples. A decrease in the crystallinity of fibers has been noted with increasing comonomer content. Relative standard deviation (R.S.D.) of the ratios were 1 and 3% for repetitive analyses on the same location and along the length of the same single fiber, respectively. When different fibers of the same sample were examined, results were reproducible within 6%. This simple method can greatly enhance the evidential value of colorless acrylic fibers, being able to discriminate them and thus helping the Court to better assess their significance.     

A simple and inexpensive molecular method for sexing and identification of the forensic samples of elephant origin

The population of the Asian elephant is being dramatically reduced due to poaching of the ivory from the male. As poaching occurs in remote forests, it often takes weeks or longer for it to be discovered and it is therefore often very difficult to determine the sex of the decomposed body. Data suggest that in the recent past, over 2000 male elephants have been poached in South India. We have developed a technique based on molecular markers to determine that the carcass is an elephant and that it is a male. Using DNA sequence information from Genbank, we have developed two primer pairs: one for the mitochondrial DNA (mtDNA) and the other for the sex-determining region of Y chromosome (SRY) gene of the Indian elephant. After PCR amplification of known elephant DNA, we found that the mtDNA was common in both males and females, whereas the SRY-specific amplicon was observed only in the male.     

Increased safety and specificity in the thin-layer chromatographic identification of marihuana

We are reporting what we believe to be a safe, rapid TLC system and spray for the identification of marihuana. The use of Fast Blue 2B salt greatly enhances the specificity of the TLC analysis of suspected marihuana samples.     

Gases released from tissue and analyzed by infrared and gas chromatography/mass spectroscopy techniques

Two techniques for analyzing contaminants released as gases from postmortem tissues were described and compared. One technique used gas chromatography/mass spectrometry (GC/MS); the other, infrared spectroscopy (IR). Brain, lung, liver, blood and urine specimens were obtained from suspected drug-overdose victims whose deaths were contributed to or caused by inhalation of unknown gases or vapors during the period immediately preceding death. Gases from the postmortem tissues and liquid samples were separately admitted into an evacuated IR gas cell, the IR spectra recorded, and gas samples then removed for GC/MS analysis. Nitrous oxide, glue, and paint solvent constituents were identified and measured. Only the brain and lung tissues contained measurable amounts of inhalants. Both IR and GC/MS methods were adequate for normal confirmatory analyses; the GC/MS system was judged superior for fast routine efforts normally hampered by incomplete sample history.     

A method for forensic identification of vegetable oil stains--rapid analysis of carboxylic acids with methyl esterification using purge-and-trap gas chromatography/mass spectrometry.

A simple method using purge-and-trap gas chromatography/mass spectrometry (P&T-GC/MS) for forensic examination of oil stains was studied. Carboxylic acids, chosen as target components for discrimination of oil samples, were extracted from stains with ether, methyl esterified by tetramethylammonium hydroxide, and analyzed by P&T-GC/MS. Vegetable oils were discriminated according to their carboxylic acid compositions. Carboxylic acid composition was independent of the substrate material of the stain. Although the carboxylic acid composition of the oil changed on exposure to sunlight, identification of oil was possible for oil stains that had been in the shade, if analysis was made within 20 days.     

Unified gas chromatography method for the detection of volatile toxicants in biological samples

A method was designed to identify and quantify the volatile toxicants in cadaveric samples; it is based on analyzing the equilibrium of the gas-vapor phase. A system comprising 3 gas chromatography columns with different polarities and without preliminary isolation of an analyzed substance is used in toxicants' identification. 0.2% n-propanol solution was used in quantification of fresh biological samples, while methanol solution was used in cases of purulence-affected materials. The method sensitivity provides for detecting some of the toxicants, like acetaldehydes, acetone as well as ethyl and higher alcohols at the level of their natural content. The method has been tested for as long as 15 years at expertise objects.     

Analytical method for the determination of strychnine in tissues by gas chromatography/mass spectrometry: two case reports

This paper describes an analytical method for strychnine determination in biological samples by gas chromatography/mass spectrometry and their application in the investigation of two cases involving strychnine ingestion: A fatal case and a clinical one. The strychnine is isolated from biological samples using a liquid-liquid extraction procedure. The clean-up procedure is performed using an acid solution. Papaverine is used as internal standard in the quantification of strychnine. In the analysed specimens, the limits of quantification were 0.1 microg/ml or 0.1 microg/g. The recovery rate ranged from 75.0% to 98.7% and the coefficients of variation ranged from 4.8% to 10.5%.     

New techniques for the identification of microscopic samples of textile fibres by infrared spectroscopy

Infrared spectroscopy has proved to be a very useful method for the routine identification of man-made textile fibres. This paper describes the way in which this technique can fit into casework procedure using two different preparative methods. Spectra can be recorded from samples of 3 μg down to less than 1 μg in weight.