Francesco Saverio Romolo

Multiclass analysis of illicit drugs in plasma and oral fluids by LC-MS/MS

AbstractAn analytical procedure for the simultaneous determination in human plasma and oral fluids of several illicit drugs belonging to different chemical and toxicological classes is presented. Amphetamine, methamphetamine, morphine, 6-monoacetylmorphine, methylenedioxyamphetamine, methylenedioxyethylamphetamine, methylenedioxymethamphetamine, cocaine, benzoylecgonine, tetrahydrocannabinol, carboxytetrahydrocannabinol, ketamine, and phencyclidine have been quantified in real samples using a very rapid sample treatment, basically a protein precipitation. The quantitative analysis was performed by liquid chromatography–tandem mass spectrometry and has been fully validated. All the analytes were detected in positive ionization mode using a TurboIonSpray source, except carboxytetrahydrocannabinol, which was detected in negative ionization mode. The use of a diverter valve between the column and the mass spectrometer allows the preservation of the ion source performances for high-throughput analysis. FigureDiverter system

Saliva and serum levetiracetam concentrations in patients with epilepsy

Abstract: Although antiepileptic drug (AED) monitoring in saliva may have some clinical applicability, it has not yet come into routine use. The correlation between levetiracetam (LEV) saliva and serum concentrations also remains unclear. To confirm LEV saliva assay as a useful, noninvasive alternative to serum measurement, we investigated the possible correlation between saliva and serum LEV concentrations. Samples of saliva and blood were collected from 30 patients with epilepsy receiving chronic therapy with LEV as monotherapy or add-on therapy, and LEV concentrations were assayed in saliva and serum. Linear regression analyses showed a close correlation between saliva and serum LEV concentrations (r2 = 0.90; P < 0.001). LEV blood and saliva concentrations were linearly related to daily drug doses (r2 = 0.78 and 0.70; P < 0.01). When data were analyzed for subgroups (patients receiving LEV in monotherapy, as add-on therapy with enzyme-inducer AEDs, and as add-on therapy with noninducer or moderate-inducer AEDs), no significant difference was found between saliva and serum LEV concentrations among groups. These preliminary results indicate that LEV, like other AEDs, can be measured in saliva as an alternative to blood-based assays. Saliva LEV collection and assay is a valid noninvasive, more convenient alternative to serum measurement.

Field detection capability of immunochemical assays during criminal investigations involving the use of TNT

The capability to collect timely information about the substances employed on-site at a crime scene is of fundamental importance during scientific investigations in crimes involving the use of explosives. TNT (2,4,6-trinitrotoluene) is one of the most employed explosives in the 20th century. Despite the growing use of improvised explosives, criminal use and access to TNT is not expected to decrease. Immunoassays are simple and selective analytical tests able to detect molecules and their immunoreactions can occur in portable formats for use on-site. This work demonstrates the application of three immunochemical assays capable of detecting TNT to typical forensic samples from experimental tests: an indirect competitive ELISA with chemiluminescent detection (CL-ELISA), a colorimetric lateral flow immunoassay (LFIA) based on colloidal gold nanoparticles label, and a chemiluminescent-LFIA (CL-LFIA). Under optimised working conditions, the LOD of the colorimetric LFIA and CL-LFIA were 1 μg mL(-1) and 0.05 μg mL(-1), respectively. The total analysis time for LFIAs was 15 min. ELISA proved to be a very effective laboratory approach, showing very good sensitivity (LOD of 0.4 ng mL(-1)) and good reproducibility (CV value about 7%). Samples tested included various materials involved in controlled explosions of improvised explosive devices (IEDs), as well as hand swabs collected after TNT handling tests. In the first group of tests, targets covered with six different materials (metal, plastic, cardboard, carpet fabric, wood and adhesive tape) were fixed on top of wooden poles (180 cm high). Samples of soil from the explosion area and different materials covering the targets were collected after each explosion and analysed. In the second group of tests, hand swabs were collected with and without hand washing after volunteers simulated the manipulation of small charges of TNT. The small amount of solution required for each assay allows for several analyses. Results of immunoassays confirmed that they were suitable to detect post-blast residues in soil and target materials and post transfer residues on hands, allowing further confirmation by more selective techniques. ELISA and LFIAs results obtained from the same solution were consistently in good agreement, and were confirmed by gas chromatography coupled to mass spectrometry (GC-MS). The reported immunoassays data demonstrates the suitability of LFIAs as on-site rapid and effective assays to detect TNT traces. The CL-ELISA proved useful in obtaining very sensitive detection in forensic investigations and testing, while CL-LFIA had performances in between LFIA and CL-ELISA.

New insights in forensic chemistry: NIR/Chemometrics analysis of toners for questioned documents examination

Near-Infrared spectroscopy (NIRs) coupled to chemometrics was investigated for the first time as a new tool for the analysis of black toners to evaluate its application in forensic cases. Ten black toners from four manufacturers were included in this study and the acquired spectra were compared in order to differentiate toners. Multivariate statistical analysis based on Principal Component Analysis (PCA) was considered to develop a model of comparison of toners in questioned documents. Results demonstrated the capabilities of the approach NIR/Chemometrics to correctly identify toners when printed on different papers and to be not affected by the printing process. This study has shown that NIRs can be considered as a useful, fast, non-destructive tool providing the characterisation of toners in forensic caseworks.

Application of micro-Raman spectroscopy for fight against terrorism and smuggling

Abstract. We report the results of Raman measurements on some common military explosives and explosives precursors deposited on clothing fabrics, both synthetic and natural, in concentration comparable to those obtained from a single fingerprint or mixed with similar harmless substances to detect illegal compounds for smuggling activities. Raman spectra were obtained using an integrated portable Raman system equipped with an optical microscope and a 785-nm laser in an analysis of <1  min. The spectral features of each illicit substance have been identified and distinguished from those belonging to the substrate fabric or from the interfering compound. Our results show that the application of Raman spectroscopy (RS) with a microscope-based portable apparatus can provide interpretable Raman spectra for a fast, in-situ analysis, directly from explosive particles of some μm3, despite the contribution of the substrate, leaving the sample completely unaltered for further, more specific, and propedeutic laboratory analysis. We also show how the RS is suitable for detecting illegal compounds mixed with harmless substances for smuggling purposes or for counterfeiting activities.

Raman spectroscopy for the detection of explosives and their precursors on clothing in fingerprint concentration: a reliable technique for security and counterterrorism issues

In this work we report the results of RS measurements on some common military explosives and some of the most common explosives precursors deposited on clothing fabrics, both synthetic and natural, such as polyester, leather and denim cotton at concentration comparable to those obtained from a single fingerprint. RS Spectra were obtained using an integrated portable Raman system equipped with an optical microscope, focusing the light of a solid state GaAlAs laser emitting at 785 nm. A maximum exposure time of 10 s was used, focusing the beam in a 45 μm diameter spot on the sample. The substances were deposited starting from commercial solutions with a Micropipetting Nano-Plotter, ideal for generating high-quality spots by non-contact dispensing of sub-nanoliter volumes of liquids, in order to simulate a homogeneous stain on the fabric surface. Images acquired with a Confocal Laser Scanning Microscope provided further details of the deposition process showing single particles of micrometric volume trapped or deposited on the underlying tissues. The spectral features of each substance was clearly identified and discriminated from those belonging to the substrate fabric or from the surrounding fluorescence. Our results show that the application of RS using a microscope-based apparatus can provide interpretable Raman spectra in a fast, in-situ analysis, directly from explosive particles of some μm3 as the ones that it could be found in a single fingerprint, despite the contribution of the substrate, leaving the sample completely unaltered for further, more specific and propaedeutic laboratory analysis. The same approach can be envisaged for the detection of other illicit substances like drugs.

Eye-safe UV Raman spectroscopy for remote detection of explosives and their precursors in fingerprint concentration

We report the results of Raman investigation performed at stand-off distance between 6-10 m with a new apparatus, capable to detect traces of explosives with surface concentrations similar to those of a single fingerprint. The device was developed as part of the RADEX prototype (RAman Detection of EXplosives) and is capable of detecting the Raman signal with a single laser shot of few ns (10-9 s) in the UV range (wavelength 266 nm), in conditions of safety for the human eye. This is because the maximum permissible exposure (MPE) for the human eye is established to be 3 mJ/cm2 in this wavelength region and pulse duration. Samples of explosives (PETN, TNT, Urea Nitrate, Ammonium Nitrate) were prepared starting from solutions deposited on samples of common fabrics or clothing materials such as blue jeans, leather, polyester or polyamide. The deposition process takes place via a piezoelectric-controlled plotter device, capable of producing drops of welldefined volume, down to nanoliters, on a surface of several cm2, in order to carefully control the amount of explosive released to the tissue and thus simulate a slight stain on a garment of a potential terrorist. Depending on the type of explosive sampled, the detected density ranges from 0.1 to 1 mg/cm2 and is comparable to the density measured in a spot on a dress or a bag due to the contact with hands contaminated with explosives, as it could happen in the preparation of an improvised explosive device (IED) by a terrorist. To our knowledge the developed device is at the highest detection limits nowadays achievable in the field of eyesafe, stand-off Raman instruments. The signals obtained show some vibrational bands of the Raman spectra of our samples with high signal-to-noise ratio (SNR), allowing us to identify with high sensitivity (high number of True Positives) and selectivity (low number of False Positives) the explosives, so that the instrument could represent the basis for an automated and remote monitoring device.

Accidental death involving professional fireworks.

An interesting case of accidental death involving the explosion of professional fireworks in an apartment is described. The examination of the scene permitted to study several effects of the explosion on walls, ceiling, furniture and especially on a balcony where the victim was found. The external examination of the victim showed extensive thermal injuries, degloving injuries and extensive shrapnel wounds. The autopsy examination showed subarachnoid haemorrhage localized to the cerebellum, haemorrhage in the soft tissues of the neck and chest and fracture of one clavicle. Almost the entire surface of lungs showed blunt injuries and the liver showed tearing of parenchyma and multiple cavities. Histological analysis were carried out showing thickening of alveolar septae, enlargement of alveolar spaces and alveolar ruptures in lung sections while numerous, round, empty spaces were detected in the parenchyma of the liver. The examination of the scene and of the fragments found showed that at least eight pyrotechnical charges exploded on the balcony, in close proximity of the threshold with the living room of the apartment. According to the chemical findings, the charges were typical for professional use and were filled with a mixture of potassium perchlorate and aluminium. A conservative calculation results in more than 1.5 kg total mass of pyrotechnic composition exploding very close to the victim.

Forensic examination of stolen-recovered vehicles: Part II: Chemical Traces—Drugs, Explosives, and Gunshot Residue

Motor vehicles are often used in the commission of crimes and are found in criminal activities, such as transporting drugs, explosives, and firearms. The detection of the presence of drugs, explosives, or Gunshot Residue (GSR) can represent very important evidence in the investigation of a crime. The three major classes of chemical traces described in this chapter could have some probative value in solving a crime: drugs, explosives, and gunshot residue. The chapter alludes to Ion Mobility Spectrometry (IMS) that is a highly sensitive analytical technique able to detect a wide range of chemical compounds (both organic and inorganic) at trace levels in the gas phase. Chemical traces can be sampled by using gas trapping technique, swabbing, vacuum lifting, or adhesive tape lifting. The chapter puts a view on the two levels of analysis for chemical traces: screening analysis and confirmatory analysis. Screening analyses are faster and cheaper than confirmatory analyses. However, screening analyses can give false-positive results, and should solely be conducted to avoid more expensive and time-consuming analysis of negative samples.

Mephedrone and Mephedrone-Based Cocktails: Market, Analysis, Mechanisms of Action, and Toxicity

Abstract In recent years mephedrone (4-methylmethcathinone) has gained popularity over other classified recreational drugs of abuse due to its low price and availability on the Internet. Because of its unwanted toxic and possibly lethal effects it is currently a matter of concern to public health worldwide. The significant qualitative and quantitative variation in content of the purportedly mephedrone-containing products on the market as well as the tendency to assume mephedrone combined with other drugs is challenging in terms of analytical identification in both street and biological samples. Furthermore, the many possible mephedrone-based combinations limit the possibilities of narrowing the clinical presentation of mephedrone intoxication, hence impairing the effectiveness of medical intervention. For these reasons awareness on the matter of mephedrone recreational use is necessary among clinicians and forensic scientists and, at least, the most common compounds derived from the combination of mephedrone and other substances should be included in toxicology screening procedures.