Search

Transient receptor potential cation channel subfamily M member 5 (TRPM5) is a nonselective monovalent cation channel activated by intracellular Ca2+ increase. Within the gastrointestinal system, TRPM5 is expressed in the stoma, small intestine, and colon. In the search for a selective agonist of TRPM5 possessing in vivo gastrointestinal prokinetic activity, a high-throughput screening was performed and compound 1 was identified as a promising hit. Hit validation and hit to lead activities led to the discovery of a series of benzo[d]isothiazole derivatives. Among these, compounds 61 and 64 showed nanomolar activity and excellent selectivity (>100-fold) versus related cation channels. The in vivo drug metabolism and pharmacokinetic profile of compound 64 was found to be ideal for a compound acting locally at the intestinal level, with minimal absorption into systemic circulation. Compound 64 was tested in vivo in a mouse motility assay at 100 mg/kg, and demonstrated increased prokinetic activity.

Metal-organic frameworks (MOFs) give the opportunity of confining guest molecules into their pores even by a post-synthetic protocol. PUM168 is a Zn-based MOF characterized by microporous cavities that allows the encapsulation of a significant number of guest molecules. The pores engineered with different binding sites show a remarkable guest affinity towards a series of natural essential oils components, such as eugenol, thymol and carvacrol, relevant for environmental applications. Exploiting single crystal X-ray diffraction, it was possible to step-wisely monitor the rather complex three-components guest exchange process involving dimethylformamide (DMF, the pristine solvent) and binary mixtures of the flavoring agents. A picture of the structural evolution of the DMF-to-guest replacement occurring inside the MOF crystal was reached by a detailed single-crystal-to-single-crystal monitoring. The relation of the supramolecular arrangement in the pores with selective guests release was then investigated as a function of time and temperature by static headspace GC-MS analysis.

The removal of toxic and carcinogenic polycyclic aromatic hydrocarbons (PAHs) from water is one of the most intractable environmental problems nowadays, because of their resistance to remediation. This work introduces a highly efficient, regenerable membrane for the removal of PAHs from water, featuring excellent filter performance and pH-driven release, thanks to the integration of a cavitand receptor in electrospun polyacrylonitrile (PAN) fibers. The role of the cavitand receptor is to act as molecular gripper for the uptake/release of PAHs. To this purpose, the deep cavity cavitand BenzoQxCav is designed and synthetized and its molecular structure is elucidated via X-Ray diffraction. The removal efficiency of the new adsorbent material toward the 16 priority PAHs is demonstrated via GC-MS analyses at ng L-1 concentration. A removal efficiency in the 32%, to 99% range is obtained. The regeneration of the membrane is performed by exploiting the pH-driven conformational switching of the cavitand between the vase form, where the PAHs uptake takes place, to the kite one, where the PAHs release occurs. The absorbance and regeneration capability of the membrane are successfully tested in four uptake/release cycles and the morphological stability.

In 2008, several synthetic cannabinoids were detected in herbal smoking blends sold on websites and in the so-called “smart shops”. These compounds, as well as new psychoactive substances, flooded the market of illicit drugs and are sold at street level. Development and validation of rapid analytical methods for the detection and quantification of synthetic cannabinoids in biological matrices are essential for the investigation of pharmacokinetic, pharmacodynamic and toxicological properties. In this pilot study, the potential of direct immersion solid-phase microextraction coupled to GC/MS for the determination of synthetic and natural cannabinoids in oral fluid was proved. Validation proved the suitability of the developed method for the determination of the analytes at trace levels in oral fluid: linearity was assessed in the LOQ – 1000 ng/mL range, whereas a good precision was observed with CV below 20%. The method was then applied to more than 100 real oral fluid samples collected from a population of young students, showing a positivity rate of 3.8%.

Particulate contamination in the form of nanoparticles (NPs) can occur not only in the environment but also in the food production chain, making human exposure almost inevitable. In the present study the potentialities of Environmental Scanning Electron Microscopy coupled with Energy-Dispersive X-ray Spectroscopy (ESEM-EDS) were exploited for the detection and visualization of inorganic NPs in air, raw materials and food products along the pasta production chain (wheat ear, wheat, semolina and pasta). Investigation of the elemental composition of NPs in the same samples was also carried out by using two independent techniques: ESEM-EDS and Inductively Coupled Plasma Mass Spectrometry (ICP-MS). As for airborne particles, size-fractionated sampling was performed by an eight-stage cascade impactor located near the production plant with different exposition times for both ESEM-EDS and gravimetric analysis, respectively. In the case of raw materials and food products, the particles were collected on polycarbonate filters after immersion in milli-Q water in order to recover the NPs deriving from external environmental contamination. Only particle sizes lower than 0.8 µm were taken into consideration when ESEM-EDS analysis was carried out. NPs containing mainly Fe and Ti having dimensions0.15 µm were identified in the filters used for wheat ear, wheat, semolina and pasta samples. As for air monitoring, gravimetric analysis showed an increase in the concentration levels of total particles during winter, particularly in the case of fine particulate matter (PM

Infections caused by the opportunistic yeast Candida albicans are one of the major life threats for hospitalized and immunocompromised patients, as a result of antibiotic and long-term antifungal treatment abuse. Odorant binding proteins can be considered interesting candidates to develop systems able to reduce the proliferation and virulence of this yeast, because of their intrinsic antimicrobial properties and complexation capabilities toward farnesol, the major quorum sensing molecule of Candida albicans. In the present study, a hybrid system characterized by a superparamagnetic iron oxide core functionalized with bovine odorant binding protein (bOBP) was successfully developed. The nanoparticles were designed to be suitable for magnetic protein delivery to inflamed areas of the body. The inorganic superparamagnetic core was characterized by an average diameter of 6.5 ± 1.1 nm and a spherical shape. Nanoparticles were functionalized by using 11-phosphonoundecanoic acid as spacer and linked to bOBP via amide bonds, resulting in a concentration level of 26.0 ± 1.2 mg bOBP/g SPIONs. Finally, both the biocompatibility of the developed hybrid system and the fungistatic activity against Candida albicans by submicromolar OBP levels were demonstrated by in vitro experiments.

Two commercial stationary phases allowing both reversed phase mechanism and anion-exchange with different selectivity, i.e. CSH C18 and Atlantis PREMIER BEH C18 AX, were tested for the separation of a complex mixture of 21 fatty acids (FAs) encompassing saturated medium-, long- and very long chain FAs, unsaturated long and very long chain FAs, cis/trans isomers, and isomers of odd- and branched-chain FAs. For this purpose, the role of surface area of stationary phase and the effect of pH of the mobile phase on the retention of the analytes were investigated. Separation was performed by ultra-high-performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS). BEH C18 AX was shown to be more versatile and to offer superior retention of these analytes to CSH C18 owing to a higher surface area and anion-exchange capacity up to pH 8.5. The UHPLC system allows shortening analysis time, the chromatographic analysis being accomplished in about 5 min, affording a high throughput of samples without the need for derivatization or ion-pairing reagents compared to techniques based upon gas chromatography approaches or LC. Finally, the application of the BEH C18 AX column using UHPLC-HRMS was demonstrated for the separation and unambiguous identification of FAs of nutritional interest in a dietary supplement sample.

A way to reduce food waste is related to the increase of the shelf-life of food as a result of improving the package type. An innovative active food packaging material based on cocrystallization of microbiologically active compounds present in essential oils i.e. carvacrol, thymol and cinnamaldehyde was developed following the Quality by Design principles. The selected active components were used to produce antimicrobial plastic films with solidified active ingredients on their surface characterized by antimicrobial properties against four bacterial strains involved in fruit and vegetable spoilage. The developed packaging prototypes exhibited good antimicrobial activity in vitro providing inhibition percentage of 69 (±15)% by contact and inhibition diameters of 32 (±6) mm in the gas phase, along with a prolonged release of the active components. Finally, the prolonged shelf-life of grape samples up to 7 days at room temperature was demonstrated.

Solid-phase microextraction and gas chromatography-mass spectrometry followed by multivariate data analysis were used to analyze lung tissues from both healthy and carcinogenic patients. A total of 78 volatile compounds belonging to different chemical classes were identified, seven of which were able to discriminate between the two groups. Discriminant analysis allowed to correctly classify 98.3% of the cases. By using the leave-one-method, 100% of the cross-validated samples belonging to the "tumor" group was correctly classified, whereas 2 cross-validated healthy samples out of 48 were erroneously allocated in the "tumor" group. Achieved results suggest the need of further investigation to assess the role of the seven identified compounds as lung cancer biomarkers in breath analysis, thus allowing the development of low-cost diagnostic devices.

During the asphalt heating process in the various stages of hot mix asphalt (HMA) production and paving operations a complex mixture of hydrocarbon aerosols, vapors and gases, generally composed of products of combustion, volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbons (PAHs), is emitted into the atmosphere. The reduction of these airborne asphalt emissions is a continuous issue and challenge for asphalt industry and road agencies. Thus, a number of new sustainable and cleaner technologies, generally referred to as warm mix asphalts (WMA), that significantly reduce the manufacturing and application temperatures, had been developed. Despite the great number of reported WMA methodologies, organic waxes are widely used for reducing the binder viscosity and for enhancing the asphalt mixture workability already at lower temperatures (90–140 °C). The manuscript presents a detailed laboratory investigation, based on headspace gas chromatography-mass spectrometry (HS-GC/MS) and photoionization detection (PID), which aimed to assess the content of VOCs and PAHs in the asphalt emissions, taking into account the effective contribution of binder or binder/wax blend. The characterization of asphalt fume composition has allowed the identification of more than 200 volatile compounds, highlighting at the same time a hierarchy of their appearance in relation to the temperature. The detection of the less volatile compounds was feasible only at temperatures higher than 160 °C, temperature range in which the HMA is generally produced. The study of the binary asphalt/wax mixtures highlighted the twofold way in which the waxes perform in reducing emissions: on the one hand they act as flow improvers (reduction of production and paving temperatures) and on the other as fume suppressants.

A new method based on microextraction by packed sorbent (MEPS) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated for the determination of dexamethasone and dexamethasone disodium phosphate in human aqueous humor. A central composite design was applied to investigate the effects of both loading and eluting cycles in the MEPS procedure; subsequently the multicriteria method of the desirability functions allowed to find the best conditions for the simultaneous extraction of both the analytes. Detection was performed on a LTQ XL linear ion trap mass spectrometer operating in the positive electrospray ionization mode applying multiple reaction monitoring mode. The assay was validated in accordance with the guidelines bioanalytical method validation obtaining quantitation limits in the low μg/L range, a precision characterized by RSD≤16% and recovery rates in the 91-119% range.

Advances in interfacing liquid chromatography and electron ionization mass spectrometry are presented. New ion source coatings synthesized by sol-gel technology were developed and tested as vaporization surfaces in terms of peak intensity, peak width and peak delay for the liquid chromatography-direct electron ionization mass spectrometry (Direct-EI) determination of environmental pollutants like polycyclic aromatic hydrocarbons and steroids. Silica-, titania-, and zirconia-based coatings were sprayed inside the stainless steel ion source and characterized in terms of thermal stability, film thickness and morphology. Negligible weight losses until 350–400 °C were observed for all the materials, with coating thicknesses in the 6 (±1)-11 (±2) μm range for optimal ionization process. The best performances in terms of both peak intensity and peak width were obtained by using the silica-based coating: the detection of the investigated compounds was feasible at low ng μl −1 levels with a good precision (RSD

In this work we report a comprehensive study leading to the fabrication of a prototype sensor for environmental benzene monitoring. The required high selectivity and ppb-level sensitivity are obtained by coupling a silicon-integrated concentration unit containing the specifically designed EtQxBox cavitand to a miniaturized PID detector. In the resulting stand-alone sensor, the EtQxBox receptor acts at the same time as highly sensitive preconcentrator for BTEX and GC-like separation phase, allowing for the selective desorption of benzene over TEX. The binding energies of the complexes between EtQxBox and BTX are calculated through molecular mechanics calculations. The examination of the corresponding crystal structures confirms the trend determined by computational studies, with the number of C-H···N and CH···? interactions increasing from 6 to 9 along the series from benzene to o-xylene. The analytical performances of EtQxBox are experimentally tested via SPME, using the cavitand as fiber coating for BTEX monitoring in air. The cavitand EFs are noticeably higher than those obtained by using the commercial CAR-DVB-PDMS. The LOD and LOQ are calculated in the ng/m range, outperforming the commercial available systems in BTEX adsorption. The desired selective desorption of benzene is achieved by applying a smart temperature program on the EtQxBox mesh, which starts releasing benzene at lower temperatures than TEX, as predicted by the calculated binding energies. The sensor performances are experimentally validated and ppb level sensitivity toward the carcinogenic target aromatic benzene was demonstrated, as required for environmental benzene exposure monitoring in industrial applications and outdoor environment.

The bacterium Pseudomonas aeruginosa (PA) and the yeast Candida albicans (CA) are pathogens that cohabit the mucosa of the respiratory tracts of animals and humans. Their virulence is largely determined by chemical communication driven by quorum sensing systems (QS), and the cross perception of their quorum sensing molecules (QSM) can modulate the prevalence of one microorganism over the other. Aiming to investigate whether some of the protein components dissolved in the mucus layering the respiratory mucosa might interfere with virulence and cross-communication of these, and eventually other microorganisms, ligand binding assays were carried out to test the scavenging potential of the bovine and porcine forms of the Lipocalin odorant binding protein (OBP) for several QSMs (farnesol, and acylhomoserine lactones), and for pyocyanin, a toxin produced by PA. In addition, the direct antimicrobial activity of the OBPs was tested by time kill assay (TKA) against CA, PA and other bacteria and yeasts. The positivity of all the ligand binding assays and the antimicrobial activity determined for CA, and for some of the other microorganisms tested, let hypothesize that vertebrate OBPs might behave as humoral components of innate immunity, active against pathogenic bacteria and fungi. In addition, TKAs with mutants of bovine OBP with structural properties different from those of the native form, and with OBP forms tagged with histidines at the amino terminal, provided information about the mechanisms responsible of their antimicrobial activity and suggested possible applications of the OBPs as alternative or co-adjuvants to antibiotic therapeutic treatments.

A reliable screening and non invasive method based on the use of microextraction by packed sorbent coupled with desorption electrospray ionization-high resolution mass spectrometry was developed and validated for the detection of new psychoactive substances in oral fluid. The role of different sample substrates in enhancing signal intensity and stability was evaluated by testing the performances of two polylactide-based materials, i.e. non-functionalized and functionalized with carbon nanoparticles, and a silica-based material compared to commercially available polytetrafluorethylene supports. The best results were achieved by using the non-functionalized polylactide substrates to efficiently ionize compounds in positive ionization mode, whereas the silica coating proved to be the best choice for operating in negative ionization mode. LLOQs in the low μg/L, a good precision with CV% always lower than 16% and RR% in the 83(±4)-120(±2)% range, proved the suitability of the developed method for the determination of the analytes in oral fluid. Finally, the method was applied for screening oral fluid samples for the presence of psychoactive substances during private parties, revealing mephedrone in only one sample out of 40 submitted to analysis.

The occurrence of emerging contaminants is becoming of increasing importance to assess the impact of anthropogenic activities onto the environment. The present study reports for the first time the development and validation of an efficient method for the simultaneous determination of fragrance materials in water samples based on the use of a novel multiwalled carbon nanotubes (MWCNTs)-based solid-phase microextraction coating. Helical MWCNTs were selected as adsorbent material due to their outstanding extraction performance. The multicriteria method of desirability functions allowed the optimization of the experimental conditions in terms of extraction time and extraction temperature. Validation proved the reliability of the method for the determination of the analytes at ultra-trace levels, obtaining detection limits in the 0.2–13 ng/L range, good precision, with relative standard deviations lower than 20% and recovery rates in the 80 ± 12%–111 ± 11%. Superior enrichment factors compared to commercial fibers were also calculated. Finally, applicability to real sample analysis was demonstrated.

The capabilities of desorption electrospray ionization-high resolution mass spectrometry (DESI-HRMS) were tested for screening the presence of some paralytic shellfish toxins in clams. A quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach is proposed for sample clean-up. QuEChERS extraction was optimized by using a full factorial design followed by the multicriteria method of the desirability functions. Quantitation limits in the microgram per kilogram range proved reliability of the method for the detection of the investigated toxins in accordance to the rules laid down by European legislation. The optimized QuEChERS-DESI-HRMS based-method allowed for a rapid reliable screening of the investigated compounds at levels of interest, thus being useful for high-throughput analyses.

Polycyclic Aromatic Hydrocarbons (PAHs) detected in polar environment are recognized tracers of anthropogenic pollution. High sensitivity and selectivity are required for their analysis in ice and snow samples due to the presence at ultra-trace levels. In this study a solid-phase microextraction (SPME) gas chromatography-mass spectrometry (GC-MS) method for simultaneous determination of the 16 US-EPA priority pollutant PAHs in polar snow and ice samples was developed and validated. Helical multiwalled carbon nanotubes (HMWCNTs) were proposed for the first time as novel SPME coating. For optimization purposes a Central Composite Design and the multicriteria method of the desirability functions were applied to investigate the influence of extraction parameters, i.e. time and temperature as main factors. The optimal values were 68 °C for the extraction temperature and 60 min for the extraction time. The developed SPME-GC-MS method exhibited detection limits of 16 PAHs in the 0.1–1.2 ng/L range, a repeatability and an intermediate precision within 15% and 22% relative standard deviation, respectively, and good recovery rates in the 93.7 (± 0.1)–119.7 (± 0.2)% range for real spiked water sample, showing better analytical performance compared to commercial PDMS fibers. Enrichment factors in the 2670 (± 290)–142120 (± 580) range were calculated and a long fiber shelf-life with the possibility to reuse the fiber more than 200 times was achieved. Finally, the proposed method was successfully applied to the determination of PAHs in surface snow samples collected in April 2019 at Ny-Alesund, Svalbard. Its application to the detection of PAHs in samples collected during monitoring campaigns in the polar regions is expected in the near future.

For the first time, a novel triple catenated Zn2+-pillared metal-organic framework (PUM 210) is proposed as solid-phase microextraction coating for the extraction of 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) from water samples and their subsequent quantitation via gas chromatography-mass spectrometry. Extraction was optimized by using a central composite face centered design followed by the multicriteria method of the desirability functions. The best conditions for the extraction of the sixteen analytes were found in correspondence to an extraction temperature of 30 °C and an extraction time of 30 min. Method validation allowed to obtain quantitation limits in the 1–7 ng/L range, a repeatability within 16% relative standard deviation (RSD), an intermediate precision within 22% RSD at limits of quantitation, and recovery rates in the 85 (±5) – 117 (±21)% range. As for selectivity, PAHs extraction proved to be feasible also in the presence of high concentration levels of potentially competing compounds like benzene, toluene, ethylbenzene and xylenes with steric hindrance playing a key role in analyte inclusion. Enrichment factors in the 300 (±5) – 14-950 (±150) range were obtained, being the heaviest compounds better extracted with respect to commercial coatings. Finally, the method was applied for the quantitation of PAHs in sixty-nine underground water samples during a monitoring campaign carried out in 2018.

In vivo solid-phase microextraction (SPME) has been recently proposed for the extraction, clean-up and preconcentration of analytes of biological and clinical concern. Bioanalysis can be performed by sampling exo- or endogenous compounds directly in living organisms with minimum invasiveness. In this context, innovative and miniaturized devices characterized by both commercial and lab-made coatings for in vivo SPME tissue sampling have been proposed, thus assessing the feasibility of this technique for biomarker discovery, metabolomics studies or for evaluating the environmental conditions to which organisms can be exposed. Finally, the possibility of directly interfacing SPME to mass spectrometers represents a valuable tool for the rapid quali- and quantitative analysis of complex matrices. This review article provides a survey of in vivo SPME applications focusing on the extraction of tissues, cells and simple organisms. This survey will attempt to cover the state-of- the-art from 2014 up to 2019.

Home-made micro-solid-phase extraction (SPE) cartridges using different adsorbent materials were tested for the desorption electrospray ionization-high-resolution mass spectrometry (DESI-HRMS) determination of explosives like 2,4,6-trinitrotoluene, cyclotrimethylene-trinitramine, cyclotetramethylene-tetranitramine, pentaerythritol tetranitrate, and trinitrophenylmethylnitramine in soil samples. Quantitation limits in the low nanogram per kilogram range proved the reliability of the method for the detection of explosives at ultra-trace levels. The reduced sample preparation allowed for low costs and high-throughput analyses. Finally, the superior extraction capability of the method was proved by obtaining DESI-HRMS responses at least five times higher than those achieved by performing DESI-HRMS analyses of solid-liquid extracts spotted onto commercial polytetrafluoroethylene slides.

Wearable solid phase microextraction (SPME) devices consisting in necklaces and pins were developed for the environmental monitoring of ketamine in recreational places using ionic liquid as coating. SPME fibers obtained using both monocationic and dicationic polymeric ionic liquids were characterized in terms of morphology, film thickness, thermal stability and pH resistance. An average thickness of 30 ± 5 μm, an excellent thermal stability until 350 °C and a very good fiber-to-fiber and batch-to-batch repeatability with RSD lower than 4% were some of the features of the developed coatings. A quantitation limit (LOQ) of 0.05 mg/m 3 with a sampling time of 1 min proved the feasibility of the developed method for the quantitation of ketamine in air at low concentration levels. Finally, the capabilities of the fibers for the rapid SPME sampling of ketamine in recreational places were proved obtaining extraction efficiencies at least two-fold higher than those obtained using commercial devices and extraction recoveries ranging from 84.2 ± 3.3% to 93.6 ± 2.6% ( n = 3).

An high performance liquid chromatography-tandem mass-spectrometry (HPLC-MS/MS) method was developed and validated for the determination in rat heart and liver of the tyrosine kinase inhibitor imatinib (IM), an anticancer drug approved for the treatment of chronic myeloid leukemia and gastrointestinal stromal tumors. Extraction of the drug from tissues was performed by solvent extraction and the obtained extracts were analyzed by HPLC-MS/MS in selected reaction monitoring mode. The developed method was validated according to the criteria for bioanalytical method, showing good performances in terms of lower limit of quantification (LLOQ=0.02μgml(-1)), linearity (R(2)=0.998), repeatability (RSD3%), reproducibility (RSD13%) and recovery (RR89%). The developed method was then applied to the analysis of heart and liver of rats treated with different doses of IM, with and without the simultaneous administration of carvedilol, a beta-blocking agent with cardioprotective effect, in order to evaluate tissue levels of the tyrosine kinase inhibitor. The obtained results revealed that the amount of IM in the rat heart was significantly affected by the administered dose, whereas carvedilol had no effect on IM concentrations. Thus, we have developed a method that allows the detection of IM traces in complex tissues such as the heart and liver and that may be proposed for the determination of the drug in other clinically relevant biological samples.

A novel series of 1,2,4-triazolyl octahydropyrrolo[2,3-b]pyrroles showing high affinity and selectivity at the DA D3 receptor is reported here. Compounds endowed with high selectivity over the hERG channel were identified and their pharmacokinetic properties thoroughly analyzed. A few derivatives with appropriate developability characteristics were selected for further studies and progression along the screening cascade. In particular, derivative 60a, (DA D3 pKi=8.4, DA D2 pKi=6.0 and hERG fpKi=5.2) showed a balanced profile and further refinements are in progress around this molecule.

Odorant binding protein (OBP) is a multi-functional scavenger for small hydrophobic molecules dissolved in the mucus lining the nasal epithelia of mammals, characterized by broad ligand binding specificity towards a large number of structurally unrelated natural and synthetic molecules of different chemical classes. Here, we demonstrate for the first time the application of OBP as the active element of an innovative filtering matrix for the removal of environmental pollutants such as triazine herbicides from water samples. The filtering device, obtained by coupling histidine-tagged bovine OBP to a nickel nitrilotriacetic acid (Ni-NTA) agarose resin, was characterized in terms of retention capacity for the herbicides atrazine, simazine, and propazine. Analysis of these herbicides at trace levels with solid-phase microextraction followed by gas chromatography-mass spectrometry using the selected ion monitoring mode proved the capabilities of the proposed device for the decontamination of surface and groundwater samples in the 0.2-2,300 μg/L concentration range, obtaining a reduction in the triazine content greater than 97 %, thus suggesting its possible use for the potabilization of water.

Raman spectroscopy followed by multivariate data analysis was used to analyze cotton fibers dyed using similar formulations and submitted to different aging conditions. Spectra were collected on a commercial instrument using a near-infrared laser with a 780nm light source. Discriminant analysis allowed to correctly classify the aged fibers 100% of the time. The prediction ability of the calculated model was estimated to be 100% by the "leave-one-out" cross-validation for 3 out of the 4 series under investigation. Finally, reliability of the developed approach for the discrimination of aged vs new fibers was confirmed by the analysis of commercial polyamide and polyester textiles submitted to the same aging process.

The study investigated the effects of choline combined with Bangerter filter in the treatment of amblyopia. All amblyopic subjects used a Bangerter filter on the corrective spectacle lens (1d over the left eye, 1d over the right eye). Choline was then administered orally to 39 patients once daily, five days per week for the entire study period. Subjects treated with the Bangerter filter showed a mean visual acuity of 0.27 logMAR; at 12mo of treatment, the mean visual acuity reached 0.09 logMAR. Patients treated with the Bangerter filter and citicoline showed a mean visual acuity of 0.35 logMAR; at 12mo of treatment, the mean visual acuity reached 0.01 logMAR. No significant changes in the angle of deviation were observed in both groups. Subjects in both forms of amblyopia therapies demonstrated an increase in visual acuity. However, these effects were markedly enhanced when coupled with the administration of choline. Findings suggest that the effects are particularly relevant in the more severe amblyopic cases.

In this study, a headspace solid-phase microextraction method was developed for the characterization of the volatile fraction of thistle honey and compared with a dynamic headspace extraction method. A DVB/CAR/PDMS fibre was used. The effects of extraction time, equilibration time and salt addition on extraction yield were evaluated. The volatile fraction of seven Italian thistle honey samples was extracted under the optimized conditions and analyzed by gas chromatography-mass spectrometry. Characterization of the volatile profile was performed in terms of nature and relative amount of the extracted compounds. A total of 40 compounds, belonging to different chemical classes, were identified. The relative amounts of 16 compounds found in all the analyzed thistle honeys, i.e. nonanal, furfural, decanal, 3,6-dimethyl-2,3,3a,4,5,7a-hexahydrobenzofuran, benzaldehyde, α-linalool, lilac aldehyde (isomer IV), hotrienol, phenylacetaldehyde, 4-oxoisophorone, benzyl alcohol, 2-phenylethanol, a not identified compound, octanoic acid, nonanoic acid and methyl anthranilate, were calculated and submitted to statistical analysis, in order to define for each compound a typical range. On the basis of the obtained data, a characteristic set of values was defined for thistle honey volatile fingerprint. The developed model proved to be effective in recognizing the botanical origin of thistle honey.

Desorption electrospray ionization mass spectrometry (DESI-MS) has been successfully used to discriminate between normal and cancerous human tissue from different anatomical sites. On the basis of this, DESI-MS imaging was used to characterize human seminoma and adjacent normal tissue. Seminoma and adjacent normal paired human tissue sections (40 tissues) from 15 patients undergoing radical orchiectomy were flash frozen in liquid nitrogen and sectioned to 15 μm thickness and thaw mounted to glass slides. The entire sample was two-dimensionally analyzed by the charged solvent spray to form a molecular image of the biological tissue. DESI-MS images were compared with formalin-fixed, hematoxylin and eosin (HE) stained slides of the same material. Increased signal intensity was detected for two seminolipids [seminolipid (16:0/16:0) and seminolipid (30:0)] in the normal tubule testis tissue; these compounds were undetectable in seminoma tissue, as well as from the surrounding fat, muscle, and blood vessels. A glycerophosphoinositol [PI(18:0/20:4)] was also found at increased intensity in the normal testes tubule tissue when compared with seminoma tissue. Ascorbic acid (i.e., vitamin C) was found at increased amounts in seminoma tissue when compared with normal tissue. DESI-MS analysis was successfully used to visualize the location of several types of molecules across human seminoma and normal tissues. Discrimination between seminoma and adjacent normal testes tubules was achieved on the basis of the spatial distributions and varying intensities of particular lipid species as well as ascorbic acid. The increased presence of ascorbic acid within seminoma compared with normal seminiferous tubules was previously unknown.

A novel diethoxydiphenylsilane-based coating for planar solid-phase microextraction was developed using sol–gel technology and used for ion mobility spectrometric detection of the explosives 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, and of the explosive taggant ethylene glycol dinitrate. The trap was characterized in terms of coating thickness, morphology, inter-batch repeatability, and extraction efficiency. An average thickness of 143 ± 13 μm with a uniform distribution of the coating was obtained. Good performances of the developed procedure in terms of both intra-batch and inter-batch repeatability with relative standard deviations

The application of an electronic nose equipped with a Metal Oxide Semiconductor sensor array for the detection of Alicyclobacillus acidoterrestris and A. acidocaldarius artificially inoculated in peach, orange and apple fruit juices is described in this study. Overall the system was able to detect the presence of Alicyclobacillus spp. in all the tested fruit juices at 24 h from inoculation. The electronic nose could detect bacterial concentration as low as

Novel series of pyrrole-pyrazinone and pyrazole-pyrazinone have been identified as potent and selective Vasopressin1b receptor antagonists. Exploration of the substitution pattern around the core of these templates allowed generation of compounds with high inhibitory potency at the Vasopressin1b receptor, including examples that showed good selectivity with respect to Vasopressin1a, Vasopressin2, and Oxytocin receptor subtypes.

A simple and easily automable method based on solid-phase microextraction followed by gas chromatographic-mass spectrometric analysis was developed for the determination of two potential angiogenesis modulators 17beta-estradiol (17-BE) and 2-methoxyestradiol (2-MEOE) in culture media. Trifluoroacetic anhydride was used as the derivatising agent. A homemade octadecyl silica coating, characterised by a coating thickness of 72 +/- 10 microm and a good thermal stability until 250 degrees C, was prepared. Experimental design was used to optimise the extraction conditions in terms of derivatisation time, derivatisation temperature and time of extraction. As for method validation, lower limits of quantification of 0.17 and 0.015 microg/l for 17beta-estradiol and 2-methoxyestradiol, respectively, were obtained. Finally, the capabilities of the developed fibres were evaluated for the analysis of the investigated analytes developed by granulosa cells in culture media maintained under normoxic, hypoxic and anoxic conditions, in order to better elucidate their possible role in the angiogenic process. An increase of the production of both 17-BE and 2-MEOE in hypoxic and anoxic conditions seems to be related to the effect of oxygen deprivation.

The optimisation of an HTS hit series (1) leading to the identification of structurally novel, selective, orally bioavailable mGluR2 positive modulators GSK1331258 and GSK1331268 is described. Structure-activity relationships, attenuation of dopaminergic activity, and potentiation of mGluR2 responses in rat hippocampal MPP-DG synapses are also reported.

An innovative and simple headspace solid-phase microextraction method using a novel diethoxydiphenylsilane fibre based on in situ derivatization with acetic anhydride was optimized and validated for the gas chromatography-mass spectrometry determination of some monohydroxy metabolites of polycyclic aromatic hydrocarbons, namely 1-hydroxynaphthalene, 2-hydroxynaphtalene, 9-hydroxyfluorene, 2-hydroxyfluorene and 1-hydroxypyrene at trace levels in human urine. Enzymatic hydrolysis was applied before derivatization, whereas extraction conditions, i.e. the effects of time and temperature of extraction and salt addition were investigated by experimental design. Regression models and desirability functions were applied to find the experimental conditions providing the highest global extraction response. These conditions were found in correspondence of an extraction temperature of 90 degrees C, an extraction time of 90 min and 25% NaCl added to urine samples. The capabilities of the developed method were proved obtaining limit of quantitations in the 0.1-2 microg/l range, thus allowing the bio-monitoring of these compounds in human urine. A good precision was observed both in terms of intra-batch and inter-batch repeatability with RSD always lower than 14%. Recoveries ranging from 98(+/-3) to 121(+/-1)% and extraction yields higher than 72% were also obtained. Finally, the analysis of urine specimens of coke-oven workers revealed analytes' concentrations in the 2.2-164 microg/l range, proving the exposure to PAHs of the involved workers.

Dynamic headspace extraction technique coupled to the gas chromatography–mass spectrometry analysis was applied to characterize volatile compounds of both crust and crumb of the Protected Designation of Origin Italian durum wheat sourdough “Altamura” bread. Volatile compounds of crust and crumb were characterized and statistically compared and their relative abundance was also calculated. A total of 89 compounds belonging to different chemical classes were identified in the crust. More abundant compounds detected in the crust were ethanol (20 ± 6%), 2-furfural (14 ± 7%) and 3-methyl-1-butanol (9 ± 5%). A lower number of volatile compounds (74) was identified in crumb, among which ethanol (32 ± 7%), 3-methyl-1-butanol (23 ± 6%) and 3-pentanol (7 ± 3) were the most abundant. The influence of different baking modes (wood- or gas-fired) on volatile compounds, macroscopic appearance and selected physico-chemical parameters (colour and texture) of bread samples were also evaluated. Samples baked in wood-fired oven showed larger amount of volatile compounds such as furans and aldehydes that could positively influence the flavour of the product. Crust of wood-fired breads showed higher amounts of compounds from Maillard reaction, resulting in harder and browner breads than gas-fired samples. Macroscopic appearance of crumb of wood-fired breads showed higher percentages of larger pores, being also less hard and cohesive than gas-fired samples.

Innovative solid-phase microextraction coatings synthesized by sol–gel technology were developed for the determination of environmental pollutants, i.e. polycyclic aromatic hydrocarbons at trace level in water. The fibers obtained, based on the use of (3-aminopropyl)triethoxysilane and diethoxydiphenylsilane were characterised in terms of film thickness, porosity, thermal stability and pH resistance. An excellent thermal stability was obtained in the case of diethoxydiphenylsilane-based coating, with a negligible weight loss until 400 °C. A very good fiber-to-fiber and batch-to-batch repeatability was also obtained with RSD lower than 6% using a mixture of aromatic hydrocarbons in the low ng/l range. Finally, the capabilities of the developed coating for sampling polycyclic aromatic hydrocarbons in water at trace levels were proved obtaining limits of detection about twofold lower than those achieved by using the commercially available polydimethylsiloxane 7 μm fiber.

A review is presented on recent applications of experimental design and optimization techniques for the analysis of compounds of food, biomedical, toxicological and environmental concern. The main features and the significant advantages of chemometric approaches are discussed. Examples related to the determination of substances like xenobiotics or naturally occurring compounds using different analytical techniques, i.e. gas chromatography (GC), liquid chromatography (LC) and capillary electrophoresis (CE) are provided in this Part. The use of experimental design techniques for optimization of extraction techniques is emphasized as well as the importance of experimental design in checking robustness of analytical methods. This survey will attempt to cover the state-of-the-art from 2004 to 2006.

A new NIR method based on multivariate calibration for determination of ethanol in industrially packed wholemeal bread was developed and validated. GC-FID was used as reference method for the determination of actual ethanol concentration of different samples of wholemeal bread with proper content of added ethanol, ranging from 0 to 3.5% (w/w) . Stepwise discriminant analysis was carried out on the NIR dataset, in order to reduce the number of original variables by selecting those that were able to discriminate between the samples of different ethanol concentrations. With the so selected variables a multivariate calibration model was then obtained by multiple linear regression. The prediction power of the linear model was optimized by a new “leave one out” method, so that the number of original variables resulted further reduced.

The capabilities of solid-phase extraction (SPE) and matrix solid-phase dispersion (MSPD) for the determination of the hormones 17β-estradiol, 2-hydroxyestradiol, 4-hydroxyestradiol and 2-methoxyestradiol by gas chromatography–mass spectrometry (GC–MS) in a very complex matrix like porcine follicular fluids were compared, thus proving the highest effectiveness of the SPE technique. Validation was carried out in terms of limit of quantitation (LOQ), precision, accuracy, recovery and stability. LOQ values in the low μg kg −1 were achieved, with all the other parameters satisfying the acceptance criteria for the validation of bioanalytical methods. The applicability of the method to the determination of the hormones in porcine follicular fluids was demonstrated, thus allowing to observe an increase of the concentration of the hormones during the follicular growth.

The capabilities of a previously developed sensor based on quartz crystal microbalance (QCM) were evaluated by using the QCM as detection unit for gas chromatography. Detection and quantitation limits towards eight compounds chosen on the basis of the coordination properties of the receptor were calculated obtaining the lowest value of limit of quantitation in the case of ethanol (LOD = 1.73 × 10 −2 μg; LOQ = 3.69 × 10 −2 μg). A novel and simple method for the determination of ethanol in bread was then developed and applied. The device proposed was proved promising as “on-line sensor” for quality control during bakery production for ethanol concentrations ranging from 0.26% to 1.70% (w/w) in whole-meal bread and from with ethanol concentration ranging from 0.68% to 2.06% (w/w) in durum-wheat bread.

Volatile compounds of roasted barley used in the production of barley coffee, the most common coffee substitute, were analysed by using solid-phase microextraction (SPME) followed by GC-MS and comprehensive GC × GC-TOF-MS, respectively. The optimised SPME extraction conditions in terms of selection of the fibre coating, extraction time and extraction temperature allowed to obtain the highest GC response, thus enhancing the identification capabilities of the developed method. As for the SPME-GC x GC-TOF-MS analysis, 64 compounds with similarity, reverse and probability values above 800, 900 and 6000, respectively, were identified, by using a polar × apolar column set combination; in contrast, GC-MS was able to identify a lower number of compounds, i. e. 40 volatiles. © 2007 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

A retention index (RI) database containing 250 volatile compounds was created on a polar stationary phase column and validated for food aroma characterisation. Precision of the retention indices (RIs) was assessed by performing replicated injections of a representative number of volatiles under the same experimental conditions: differences lower than 1 U were observed for all the compounds. Robustness was evaluated by carrying out injections of the same set of volatile compounds under different experimental conditions, i. e. program temperature, column batches and instrumentation. Excellent results were obtained with a maximum difference in the RI values of 10 U. The capabilities of the created database for food aroma characterisation were finally evaluated by analysing the volatile fractions of different food matrices such as dry sausages, cheese and bread. A great number of volatile compounds were identified in the analysed samples on the basis of their RI, thus proving the usefulness of the RI collections in the field of food analysis.

An acetone extract of Helichrysum italicum ssp. microphyllum afforded the phloroglucinol alpha-pyrone arzanol (1a) as a potent NF-kappaB inhibitor. Arzanol is identical with homoarenol (2a), whose structure should be revised. The phloroglucinol-type structure of arzanol and the 1,2,4-trihydroxyphenyl-type structure of the base-induced fragmentation product of homoarenol could be reconciled in light of a retro-Fries-type fragmentation that triggers a change of the hydroxylation pattern of the aromatic moiety. On the basis of these findings, the structure of arenol, the major constituent of the clinically useful antibiotic arenarin, should be revised from 2b to 1b, solving a long-standing puzzle over its biogenetic derivation. An alpha-pyrone (micropyrone, 7), the monoterpene rac-E-omega-oleoyloxylinalol (10), four known tremetones (9a-d), and the dimeric pyrone helipyrone (8) were also obtained. Arzanol inhibited HIV-1 replication in T cells and the release of pro-inflammatory cytokines in LPS-stimulated primary monocytes, qualifying as a novel plant-derived anti-inflammatory and antiviral chemotype worth further investigation.