Chitosan films have attracted increased attention in the field of sensors because of chitosan’s u... more Chitosan films have attracted increased attention in the field of sensors because of chitosan’s unique chemico-physical properties, including high adsorption capacity, filmability and transparency. A chitosan film sensor was developed through the dispersion of an ammonia specific reagent (Nessler’s reagent) into a chitosan film matrix. The chitosan film sensor was characterized to assess the film’s properties by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). A gas diffusion device was prepared with the chitosan film sensor, enabling the collection and detection of ammonia vapor from biological samples. The chitosan film sensor color change was correlated with the ammonia concentration in samples of human serum and artificial urine. This method enabled facile ammonia detection and concentration measurement, making the sensor useful not only in clinical laboratories, but ...
A paper microfluidic device (microPAD) has been developed for the presumptive testing of seized d... more A paper microfluidic device (microPAD) has been developed for the presumptive testing of seized drugs. The microPAD was composed by hydrophilic channels on chromatographic paper by means wax printing and thermal lamination. Six different lanes were connected to a single stem. A different colorimetric reaction occurred within each lane, permitting the multiplexed detection of different substances including cocaine, opiates, ketamine, and various phenethylamines. Furthermore, the orientation allowed for using series of reactants in each channel, enhancing stability and permitting a sequential interaction with the analyte. The microPAD was evaluated for sensitivity and tested with common interferences and drug diluents. A further development of this approach for drug detecting was the coupling of the paper microfluidic device with gold nanoparticles and anti-cocaine aptamers to make the detection of cocaine more sensitive and specific. The operation of the device is easy and rapid, and does not require extensive training or instrumentation.\ua0In addition, all of the materials used in the device are safe and environmentally friendly.\ua0For these reasons, the microPAD should prove a useful tool for the screening of seized drugs
The fast paced advancement in molecularly imprinted nanoparticles (MIP NPs) synthesis and applica... more The fast paced advancement in molecularly imprinted nanoparticles (MIP NPs) synthesis and applications [1] requires straightforward analytical methods suitable for the evaluation of the binding properties of the generated MIP NP libraries. Often the analysis of the MIP NP binding behaviour cannot be easily performed by classical means, because of their nature, i.e. amphipathic, polymeric and their dimensions (10-200 nm), which are greater than biological macromolecules, but smaller than most of the micromaterials. In response to these needs, capillary electrophoresis (CE) was here exploited to screen the binding affinities of MIP NPs targeting the iron regulating hormone hepcidin towards their template peptide, i.e. the N terminal 6-mer of hepcidin [2- 4]. The CE separation method was developed ex novo, after optimization of the background electrolyte (150 mM sodium phosphate pH 7.4) and of the running temperature (35\ub0C), achieving the full separation of the free ligand from the complexed MIP NPs. The CE binding isotherm allowed estimating a micromolar dissociation constant for the 6-mer template/MIP NP complex. The results were found in agreement with independent measures. The CE offers the advantages of a direct analysis of the MIP NP/ligand incubation mix without preliminary fractionation steps, requiring only minimal sample consumption and short analysis times, thus in conclusion, it appeared to be a valid technique for characterizing the interaction of MIP NP libraries for selected target compounds
The penetration of the new psychoactive substances (NPSs) into the market of clandestine drugs is... more The penetration of the new psychoactive substances (NPSs) into the market of clandestine drugs is highly dynamic, causing potentially false-negative results using the current analytical instrumentation, particularly in the screening phase. At present, the suggested approach to perform a comprehensive screening requires the use of high-resolution mass spectrometry (MS) with associated high costs of purchase and maintenance and need of skilled and dedicated personnel. Here we describe the development and validation of a simplified approach based on a high-performance liquid chromatography–ion trap MS system with a user-friendly interface dedicated to toxicological analysis. The system, originally intended for a broad toxicological screening, was tuned to identify new synthetic cannabinoids in hair. After a washing step with dichloromethane, hair (about 50 mg) was incubated for 3 h with 1.5 mL ethanol. One milliliter of this solution was then dried, reconstituted with mobile phase and injected. The peak identification was based on the chromatographic retention times and MS2/MS3 data using a database which included up to 158 NPSs. The method was validated according to international guidelines on a selected panel of NPSs, namely methyl 2-[[1-(5-fluoropentyl)indazole-3-carbonyl]amino]-3,3-dimethylbutanoate (5F-ADB), 1-Pentyfluoro-1H-indole-3-carboxylic acid 8-quinolinyl ester (5F-PB 22), N-[(2S)-1-amino-3-methyl-1-oxobutan-2-yl]-1-(5-chloropentyl)indazole-3-carboxamide (5Cl-AB-PINACA), (S)-N-(1-amino-1-oxo-3-phenylpropan-2-yl)-1-(5-fluoropentyl)-1H-indole-3-carboxamide [5F-APP-PICA (PX-1)],: (R)-N-(1-amino-1-oxo-3-phenylpropan-2-yl)-1-(5-fluoropentyl)-1H-indazole-3-carboxamide [5F-APP-PINACA (PX-2)], N-[(2S)-1-Amino-3-methyl-1-oxobutan-2-yl]-1-(cyclohexylmethyl)indazole-3-carboxamide (AB-CHMINACA), N-[(2S)-1-Amino-3-methyl-1-oxobutan-2-yl]-1-[(4-fluorophenyl)methyl] indazole-3-carboxamid (AB-FUBINACA), methyl (2S)-2-[[1-(cyclohexylmethyl)indole-3-carbonyl]amino]-3,3-dimethylbutanoate (MDMB-CHMICA), (S)-Methyl 2-(1-(5-fluoropentyl)-1H-indole-3-carboxamido)-3-methylbutanoate (MMB-2201) and (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone (UR-144). The tested analytical method showed detection limits between 0.065 and 0.125 ng/mg. The intraday imprecision of the method showed average values within the range of 7.3–20%. The estimation of the trueness (bias) of method showed average values within the range of 1.5–12.3%. The analytical performance was also successfully assessed by four proficiency test samples containing NPS. No synthetic cannabinoids were detected in application to 82 hair samples from forensic cases previously analyzed with liquid chromatography–MS triple quadrupole.
According to the World Health Organization, as many as 25% of traffic accidents are linked to alc... more According to the World Health Organization, as many as 25% of traffic accidents are linked to alcohol abuse. This study describes the results of a nine-year study performed on injured drivers ( N = 12,806) in the Verona area of Northern Italy. Blood samples were mandatorily collected on injured drivers who were admitted to the Emergency Health Care Unit of Verona Hospital between 2009 and 2017, after they had been involved in a traffic accident. Blood alcohol concentration (BAC) determination was then undertaken using a validated head space–gas chromatography–flame ionisation detector (HS-GC-FID) method. We found that 21% of drivers tested positive for alcohol (BAC ≥0.01 g/L), while 16.8% presented with BAC levels above the Italian legal limit (>0.5 g/L). Of those who had positive BACs, about 50% presented with very high BAC levels (>1.5 g/L). Daily time distribution analyses, involving 2031 alcohol-positive drivers, showed a surge between 18:00 hours and 06:00 hours (74.3%), with a specific rise during the weekend (58.9%). The percentage of alcohol-related road accidents was 20.6%, which is lower than results reported in other international studies performed over the last 20 years. However, evidence that around 50% of the positive subjects showed a BAC >1.5 g/L confirms the correlation between BAC and accident risk, which becomes even more significant at progressively increasing levels of BAC. The study highlights the need to implement further strategies to both prevent and deter the use of alcohol while driving.
The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxic... more The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxication has prompted researchers to develop rapid, sensitive, and cost-effective methods and analytical devices suitable for use in nonspecialized laboratories and at the point of need (PON). In recent years, the technology of paper-based microfluidic analytical devices (μPADs) has undergone rapid development and now provides a feasible, low-cost alternative to traditional rapid tests for detecting harmful compounds. In fact, µPADs have been developed to detect toxic molecules (arsenic, cyanide, ethanol, and nitrite), drugs, and drugs of abuse (benzodiazepines, cathinones, cocaine, fentanyl, ketamine, MDMA, morphine, synthetic cannabinoids, tetrahydrocannabinol, and xylazine), and also psychoactive substances used for drug-facilitated crimes (flunitrazepam, gamma-hydroxybutyric acid (GHB), ketamine, metamizole, midazolam, and scopolamine). The present report critically evaluates the recent...
Chitosan films have attracted increased attention in the field of sensors because of chitosan’s u... more Chitosan films have attracted increased attention in the field of sensors because of chitosan’s unique chemico-physical properties, including high adsorption capacity, filmability and transparency. A chitosan film sensor was developed through the dispersion of an ammonia specific reagent (Nessler’s reagent) into a chitosan film matrix. The chitosan film sensor was characterized to assess the film’s properties by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). A gas diffusion device was prepared with the chitosan film sensor, enabling the collection and detection of ammonia vapor from biological samples. The chitosan film sensor color change was correlated with the ammonia concentration in samples of human serum and artificial urine. This method enabled facile ammonia detection and concentration measurement, making the sensor useful not only in clinical laboratories, but ...
A paper microfluidic device (microPAD) has been developed for the presumptive testing of seized d... more A paper microfluidic device (microPAD) has been developed for the presumptive testing of seized drugs. The microPAD was composed by hydrophilic channels on chromatographic paper by means wax printing and thermal lamination. Six different lanes were connected to a single stem. A different colorimetric reaction occurred within each lane, permitting the multiplexed detection of different substances including cocaine, opiates, ketamine, and various phenethylamines. Furthermore, the orientation allowed for using series of reactants in each channel, enhancing stability and permitting a sequential interaction with the analyte. The microPAD was evaluated for sensitivity and tested with common interferences and drug diluents. A further development of this approach for drug detecting was the coupling of the paper microfluidic device with gold nanoparticles and anti-cocaine aptamers to make the detection of cocaine more sensitive and specific. The operation of the device is easy and rapid, and does not require extensive training or instrumentation.\ua0In addition, all of the materials used in the device are safe and environmentally friendly.\ua0For these reasons, the microPAD should prove a useful tool for the screening of seized drugs
The fast paced advancement in molecularly imprinted nanoparticles (MIP NPs) synthesis and applica... more The fast paced advancement in molecularly imprinted nanoparticles (MIP NPs) synthesis and applications [1] requires straightforward analytical methods suitable for the evaluation of the binding properties of the generated MIP NP libraries. Often the analysis of the MIP NP binding behaviour cannot be easily performed by classical means, because of their nature, i.e. amphipathic, polymeric and their dimensions (10-200 nm), which are greater than biological macromolecules, but smaller than most of the micromaterials. In response to these needs, capillary electrophoresis (CE) was here exploited to screen the binding affinities of MIP NPs targeting the iron regulating hormone hepcidin towards their template peptide, i.e. the N terminal 6-mer of hepcidin [2- 4]. The CE separation method was developed ex novo, after optimization of the background electrolyte (150 mM sodium phosphate pH 7.4) and of the running temperature (35\ub0C), achieving the full separation of the free ligand from the complexed MIP NPs. The CE binding isotherm allowed estimating a micromolar dissociation constant for the 6-mer template/MIP NP complex. The results were found in agreement with independent measures. The CE offers the advantages of a direct analysis of the MIP NP/ligand incubation mix without preliminary fractionation steps, requiring only minimal sample consumption and short analysis times, thus in conclusion, it appeared to be a valid technique for characterizing the interaction of MIP NP libraries for selected target compounds
The penetration of the new psychoactive substances (NPSs) into the market of clandestine drugs is... more The penetration of the new psychoactive substances (NPSs) into the market of clandestine drugs is highly dynamic, causing potentially false-negative results using the current analytical instrumentation, particularly in the screening phase. At present, the suggested approach to perform a comprehensive screening requires the use of high-resolution mass spectrometry (MS) with associated high costs of purchase and maintenance and need of skilled and dedicated personnel. Here we describe the development and validation of a simplified approach based on a high-performance liquid chromatography–ion trap MS system with a user-friendly interface dedicated to toxicological analysis. The system, originally intended for a broad toxicological screening, was tuned to identify new synthetic cannabinoids in hair. After a washing step with dichloromethane, hair (about 50 mg) was incubated for 3 h with 1.5 mL ethanol. One milliliter of this solution was then dried, reconstituted with mobile phase and injected. The peak identification was based on the chromatographic retention times and MS2/MS3 data using a database which included up to 158 NPSs. The method was validated according to international guidelines on a selected panel of NPSs, namely methyl 2-[[1-(5-fluoropentyl)indazole-3-carbonyl]amino]-3,3-dimethylbutanoate (5F-ADB), 1-Pentyfluoro-1H-indole-3-carboxylic acid 8-quinolinyl ester (5F-PB 22), N-[(2S)-1-amino-3-methyl-1-oxobutan-2-yl]-1-(5-chloropentyl)indazole-3-carboxamide (5Cl-AB-PINACA), (S)-N-(1-amino-1-oxo-3-phenylpropan-2-yl)-1-(5-fluoropentyl)-1H-indole-3-carboxamide [5F-APP-PICA (PX-1)],: (R)-N-(1-amino-1-oxo-3-phenylpropan-2-yl)-1-(5-fluoropentyl)-1H-indazole-3-carboxamide [5F-APP-PINACA (PX-2)], N-[(2S)-1-Amino-3-methyl-1-oxobutan-2-yl]-1-(cyclohexylmethyl)indazole-3-carboxamide (AB-CHMINACA), N-[(2S)-1-Amino-3-methyl-1-oxobutan-2-yl]-1-[(4-fluorophenyl)methyl] indazole-3-carboxamid (AB-FUBINACA), methyl (2S)-2-[[1-(cyclohexylmethyl)indole-3-carbonyl]amino]-3,3-dimethylbutanoate (MDMB-CHMICA), (S)-Methyl 2-(1-(5-fluoropentyl)-1H-indole-3-carboxamido)-3-methylbutanoate (MMB-2201) and (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone (UR-144). The tested analytical method showed detection limits between 0.065 and 0.125 ng/mg. The intraday imprecision of the method showed average values within the range of 7.3–20%. The estimation of the trueness (bias) of method showed average values within the range of 1.5–12.3%. The analytical performance was also successfully assessed by four proficiency test samples containing NPS. No synthetic cannabinoids were detected in application to 82 hair samples from forensic cases previously analyzed with liquid chromatography–MS triple quadrupole.
According to the World Health Organization, as many as 25% of traffic accidents are linked to alc... more According to the World Health Organization, as many as 25% of traffic accidents are linked to alcohol abuse. This study describes the results of a nine-year study performed on injured drivers ( N = 12,806) in the Verona area of Northern Italy. Blood samples were mandatorily collected on injured drivers who were admitted to the Emergency Health Care Unit of Verona Hospital between 2009 and 2017, after they had been involved in a traffic accident. Blood alcohol concentration (BAC) determination was then undertaken using a validated head space–gas chromatography–flame ionisation detector (HS-GC-FID) method. We found that 21% of drivers tested positive for alcohol (BAC ≥0.01 g/L), while 16.8% presented with BAC levels above the Italian legal limit (>0.5 g/L). Of those who had positive BACs, about 50% presented with very high BAC levels (>1.5 g/L). Daily time distribution analyses, involving 2031 alcohol-positive drivers, showed a surge between 18:00 hours and 06:00 hours (74.3%), with a specific rise during the weekend (58.9%). The percentage of alcohol-related road accidents was 20.6%, which is lower than results reported in other international studies performed over the last 20 years. However, evidence that around 50% of the positive subjects showed a BAC >1.5 g/L confirms the correlation between BAC and accident risk, which becomes even more significant at progressively increasing levels of BAC. The study highlights the need to implement further strategies to both prevent and deter the use of alcohol while driving.
The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxic... more The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxication has prompted researchers to develop rapid, sensitive, and cost-effective methods and analytical devices suitable for use in nonspecialized laboratories and at the point of need (PON). In recent years, the technology of paper-based microfluidic analytical devices (μPADs) has undergone rapid development and now provides a feasible, low-cost alternative to traditional rapid tests for detecting harmful compounds. In fact, µPADs have been developed to detect toxic molecules (arsenic, cyanide, ethanol, and nitrite), drugs, and drugs of abuse (benzodiazepines, cathinones, cocaine, fentanyl, ketamine, MDMA, morphine, synthetic cannabinoids, tetrahydrocannabinol, and xylazine), and also psychoactive substances used for drug-facilitated crimes (flunitrazepam, gamma-hydroxybutyric acid (GHB), ketamine, metamizole, midazolam, and scopolamine). The present report critically evaluates the recent...
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