Gabinetto Regionale di Polizia Scientifica per lEmilia Romagna

Bologna, Italy

Gabinetto Regionale di Polizia Scientifica per lEmilia Romagna

Bologna, Italy
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Girotti S.,University of Bologna | Eremin S.,Moscow State University | Montoya A.,Polytechnic University of Valencia | Moreno M.J.,Polytechnic University of Valencia | And 6 more authors.
Analytical and Bioanalytical Chemistry | Year: 2010

To identify the explosive used in a terrorist attack, or to obtain an early sign of environmental pollution it is important to use simple and rapid assays able to detect analytes at low levels, possibly on-site. This is particularly true for TNT (2,4,6-trinitrotoluene), one of the most employed explosives in the 20th century and at the same time, because of its toxicity, a well known pollutant. In this work we describe the development of an indirect competitive ELISA with chemiluminescent detection (CL-ELISA) and of a lateral-flow immunoassay (LFIA) based on colloidal gold nanoparticle labels. A commercially available monoclonal antibody was used and 13 specially synthesized conjugates were tested. We optimized the assay by determining the optimal concentration of monoclonal antibody and conjugates and the influence of various non-specific factors such as: tolerance to organic solvents at different concentrations, the washing and competitive step time, and the cross-reactivity with related compounds. The sensitivity and reproducibility of the CL-ELISA were good (LOD and IC50 values in the ng mL-1 range, and CV value about 7%). It has been applied to real samples of various materials involved in a controlled explosion of an "improvised explosive device". Three extraction procedures were tested on these samples, all employing methanol as the solvent. The lateral flow immunoassay (LFIA), developed by using the same immunoreagents, reached a detection limit of 1 μg mL-1 when tested on the same samples analysed by CL-ELISA. © 2009 Springer-Verlag.

Cevenini L.,Italian National Institute of Biosystems and Biostructures | Michelini E.,Italian National Institute of Biosystems and Biostructures | Michelini E.,University of Bologna | D'Elia M.,Gabinetto Regionale di Polizia Scientifica per lEmilia Romagna | And 3 more authors.
Analytical and Bioanalytical Chemistry | Year: 2013

Bioassays represent promising complementary techniques to conventional analytical approaches used in doping analysis to detect illicit drugs like anabolic-androgenic steroids (AAS). The fact that all AAS share a common mechanism of action via the human androgen receptor (hAR) enables the use of bioassays, relying on the activation of hAR as antidoping screening tools. Previously, we developed a dual-color bioreporter based on yeast cells engineered to express hAR and androgen response elements driving the expression of the bioluminescent (BL) reporter protein Photinus pyralis luciferase. A second reporter protein, the red-emitting luciferase PpyRE8, was introduced in the bioreporter as internal viability control. Here, we report the first forensic application of a straightforward, accurate, and cost-effective bioassay, relying on spectral resolution of the two BL signals, in 96-microwell format. The bioreporter responds to dihydrotestosterone as reference androgen in a concentration-dependent manner from 0.08 to 1,000 nM with intra- and inter-assay variation coefficients of 11.4 % and 13.1 %, respectively. We also demonstrated the suitability of this dual-color bioreporter to assess (anti)-androgenic activity of pure AAS, mixtures of AAS, and other illicit drugs provided by the Scientific Police. Significant anti-androgenic activity was observed in samples labeled as marijuana and hashish, containing Δ9-tetrahydrocannabinol as major constituent. [Figure not available: see fulltext.] © 2012 Springer-Verlag.

Girotti S.,University of Bologna | Ferri E.,University of Bologna | Maiolini E.,University of Bologna | Bolelli L.,University of Bologna | And 4 more authors.
Analytical and Bioanalytical Chemistry | Year: 2011

A quantitative chemiluminescent method, enabling indirect identification of the peroxide-based explosives TATP (triacetone triperoxide) and HMTD (hexamethylene triperoxide diamine) has been developed. Treatment of these compounds with acidic solutions produced peroxides, which were transformed into radical derivatives by horseradish peroxidase (HRP) and then quantified by measuring the light emitted during their oxidation of luminol. The method was first developed in the microplate format and later optimized for a portable luminometer, to enable rapid application of the assay directly on site. When the portable luminometer was used each analysis took only 5-10 min. The method had good selectivity, sensitivity, and reproducibility; in the microplate format the limits of detection (LOD) and quantification (LOQ) were 40 and 50 ng mL -1, respectively, for both TATP and HMTD. When the portable luminometer was used the LOD and LOQ were 50 and 100 ng mL -1, respectively, for both compounds. Introduction of light emission-enhancing compounds did not improve the analytical performance of the assay. Imprecision (CV values) was always below 10%. Recovery varied rapidly with time, with an average value of 78% after 5 min. No false-positive result was detected on measurement of a variety of samples; this is an important feature for analysis on site. The method was applied both to contaminated materials and to fortified soil samples, simulating operational conditions. © 2011 Springer-Verlag.

Valle F.,CNR Institute of Nanostructured Materials | Bianchi M.,CNR Institute of Nanostructured Materials | Tortorella S.,CNR Institute of Nanostructured Materials | Pierini G.,University of Bologna | And 2 more authors.
Forensic Science International | Year: 2012

A novel application of replica molding to a forensic problem, viz. the accurate reproduction of the case head of gun and rifle cartridges, prior and after been shot, is presented. The fabrication of an arbitrary number of identical copies of the region hit by the firing pin and by the breech face is described. The replicas can be (i) handled without damaging the original evidence, (ii) distributed to different law enforcement agencies for comparison against other evidences found on crime scenes or ballistic tests of seized firearms, (iii) maintained on a file by the laboratories. A detailed analysis of the morphological features of the replicas has been carried out by standard microscopy techniques as well as by advanced microscopy such as scanning probe and scanning electron leading to a quantitative morphological characterization of the case heads down to the nanometer scale. The assignment of the cartridge replicas to the shooting weapon is demonstrated to hold below the micron scale, while it is hindered at the nanometer level both by the manufacturing differences and by eventual modifications occurring on the firing pin. © 2012 Elsevier Ireland Ltd.

Cominato L.,CNR Institute of Nanostructured Materials | Valle F.,CNR Institute of Nanostructured Materials | Pierini G.,University of Bologna | Bonini P.,Gabinetto Regionale di Polizia Scientifica per lEmilia Romagna | And 2 more authors.
Forensic Science International | Year: 2015

The application of replica molding has proven to be a valuable tool in the analysis of different forensic evidences in particular for its ability to extract the toolmarks from complex sample surfaces. A well known problem in the analysis of ballistic evidences is the accurate characterization of the lateral striae of real bullets seized on crime scenes after shots, due primarily to impact deformations and to unpredictable issues related to laboratory illumination setup. To overcome these problems a possible way is to confine over a flat surface all the features still preserving their three dimensionality. This can be achieved by a novel application of replica molding performed onto the relevant lateral portion of the bullet surface. A quasi-two-dimensional negative copy of the original tridimensional indented surface has been thus fabricated. It combines the real tridimensional topography of class characteristics (land and groove impressions) and of individual caracteristics (striae) impressed by rifled barrels on projectiles, moreover with the possibility of quantitative characterization of these features in a planar configuration, that will allow one-shot comparison of the "whole striae landscape" without the typical artifacts arising from the bullet shape and the illumination issue. A detailed analysis has been carried on at the morphological level by standard optical and scanning electron microscopy, while the 3D topography has been characterized by white light optical profilometry. A quantitative characterization of toolmarks of bullets derived from ammunitions shot by guns of large diffusion, as the Beretta 98 FS cal. 9 × 21 mm, has been performed and will be presented ranging between the whole landscape and the sub-μm resolution. To investigate the real potentiality of this technique, the experiment has been extended to highly impact-deformed projectiles. © 2014 Elsevier Ireland Ltd.

Melucci D.,University of Bologna | Monti D.,University of Bologna | D'Elia M.,Gabinetto Regionale di Polizia Scientifica per lEmilia Romagna | Luciano G.,National Research Council Italy
Journal of Forensic Sciences | Year: 2012

This study takes the first step toward in situ analysis of powder drugs which does not require any alteration of the samples. A fast, inexpensive analytical method based on reflectance near-infrared (NIR) spectrometry and multivariate calibration was applied. A diode-array fiber-optic portable spectrometer in the 900-1700nm range was employed. Samples were laboratory-prepared ternary powders (diacetylmorphine, caffeine, and paracetamol). Partial least squares regression was applied. The choice of the standard samples for calibration and validation was performed through a D-optimal experimental design. The explained variance was higher than 90%, and the relative root mean square errors were <2%. The number of principal components (6) was very low when compared with the number of raw variables (356 absorbance values). Response plots showed slopes and intercepts were very close to optimal values. Correlation coefficients ranged between 0.909 and 0.989. The method here proposed proved to be competitive with Fourier transform NIR spectrometry. © 2011 American Academy of Forensic Sciences.

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