Application of laser ablation multicollector inductively coupled plasma mass spectrometry for the measurement of calcium and lead isotope ratios in packaging for discriminatory purposes

The potential of high‐precision calcium and lead isotope ratio measurements using laser ablation coupled to multicollector inductively coupled plasma mass spectrometry (LA‐MC‐ICP‐MS) to aid distinction between four genuine and five counterfeit pharmaceutical packaging samples and further classification of counterfeit packaging samples has been evaluated. We highlight the lack of reference materials for LA‐MC‐ICP‐MS isotope ratio measurements in solids. In this case the problem is minimised by using National Institute of Standards and Technology Standard Reference Material (NIST SRM) 915a calcium carbonate (as solid pellets) and NIST SRM610 glass disc for sample bracketing external standardisation. In addition, a new reference material, NIST SRM915b calcium carbonate, has been characterised in‐house for Ca isotope ratios and is used as a reference sample. Significant differences have been found between genuine and counterfeit samples; the method allows detection of counterfeits and aids further classification of packaging samples. Typical expanded uncertainties for measured‐corrected Ca isotope ratio values (⁴³Ca/⁴⁴Ca and ⁴²Ca/⁴⁴Ca) were found to be below 0.06% (k = 2, 95% confidence) and below 0.2% for measured‐corrected Pb isotope ratios (²⁰⁷Pb/²⁰⁶Pb and ²⁰⁸Pb/²⁰⁶Pb). This is the first time that Ca isotope ratios have been measured in packaging materials using LA coupled to a multicollector (MC)‐ICP‐MS instrument. The use of LA‐MC‐ICP‐MS for direct measurement of Ca and Pb isotopic variations in cardboard/ink in packaging has definitive potential to aid counterfeit detection and classification. Copyright © 2010 John Wiley & Sons, Ltd.     

QSAR classification and regression models for β-secretase inhibitors using relative distance matrices

The development of robust QSAR models to predict the activity of molecules of β-secretase inhibitors is an area of interest due to the increase of Alzheimer’s disease in patients in the global population. In this paper, we present a proposal based on the use of relative distance matrices as input data to the QSAR algorithms. These matrices store measurements of distances between the structural characteristics of pairs of molecules and between the molecules and a structural pattern extracted from the whole data set, thus efficiently representing a correlation between structural changes and activity. For the building of the classification and regression models support vector machine, tree complex and Gaussian process algorithms have been used; and for the validation of the models cross-validation, bootstrapping and y-randomizing techniques have been applied. The results obtained are close to 100% in accuracy and area under receiver operating characteristic values in classification, and close to 1.0 for r ² and 0.1 for root mean square error in regression in training and in external validation, proving the ‘goodness’ of the proposal.     

Development and validation of a ultra-high-performance liquid chromatography-UV method for the detection and quantification of erectile dysfunction drugs and some of their analogues found in counterfeit medicines

Pharmaceutical counterfeiting is a permanently growing problem. Control laboratories are constantly analysing counterfeit medicines. In industrialised countries, one of the main counterfeited class of medicines are erectile dysfunction drugs. This paper describes the development and validation of a fast method to detect and quantify the three authorised phosphodiesterase type 5 inhibitors and five analogues. The method is based on the use of a sub-2 microns polar-embedded column with a gradient using acetonitrile as organic modifier and 10 mM ammonium formate buffer (pH 3.5) as aqueous component of the mobile phase. The separation was achieved in less than 4.5 min. The method has also been compared to the registered HPLC method for the assay of Viagra^® which was considered as the reference method. The method is also compatible with on-line coupling mass spectrometry and will significantly reduce analysis times and solvent consumption.     

Classification of Greek Olive Oils from Different Regions by Machine Learning-Aided Laser-Induced Breakdown Spectroscopy and Absorption Spectroscopy

In the present work, the emission and the absorption spectra of numerous Greek olive oil samples and mixtures of them, obtained by two spectroscopic techniques, namely Laser-Induced Breakdown Spectroscopy (LIBS) and Absorption Spectroscopy, and aided by machine learning algorithms, were employed for the discrimination/classification of olive oils regarding their geographical origin. Both emission and absorption spectra were initially preprocessed by means of Principal Component Analysis (PCA) and were subsequently used for the construction of predictive models, employing Linear Discriminant Analysis (LDA) and Support Vector Machines (SVM). All data analysis methodologies were validated by both “k-fold” cross-validation and external validation methods. In all cases, very high classification accuracies were found, up to 100%. The present results demonstrate the advantages of machine learning implementation for improving the capabilities of these spectroscopic techniques as tools for efficient olive oil quality monitoring and control.     

Near-infrared imaging for quantitative analysis of active component in counterfeit dimethomorph using partial least squares regression

Near-infrared (NIR) imaging systems simultaneously record spectral and spatial information. Near-infrared imaging was applied to the identification of (E,Z)-4-(3-(4-chlorophenyl)-3-(3,4-dimethoxyphenyl)acryloyl)morpholine (dimethomorph) in both mixed samples and commercial formulation in this study. The distributions of technical dimethomorph and additive in the heterogeneous counterfeit product were obtained by the relationship imaging (RI) mode. Furthermore, a series of samples which consisted of different contents of uniformly distributed dimethomorph were prepared and three data cubes were generated for each content. The spectra extracted from these images were imported to establish the partial least squares model. The model??s evaluating indicators were: coefficient of determination (R ²) 99.42 %, root mean square error of calibration (RMSEC) 0.02612, root mean square error of cross-validation (RMSECV) 0.01693, RMSECV_mean 0.03577, relative standard error of prediction (RSEP) 0.01999, and residual predictive deviation (RPD) 15.14. Relative error of prediction of the commercial formulation was 0.077, indicating the predicted value correlated with the real content. The chemical value reconstruction image of dimethomorph formulation products was calculated by a MATLAB program. NIR microscopy imaging here manifests its potential in identifying the active component in the counterfeit pesticide and quantifying the active component in its scanned image.     

Exchange properties of cyanide complexes

In the year 2000, at the MARC V conference, the first results obtained at constant count rates with so-called "zero dead time counting" (ZDT) as implemented in ORTEC's DSPEC^{PLUS ®} were presented. In this paper, further experiments are described that were performed to establish how the DSPEC^{PLUS ®} performs at varying count rates. At the same time, the experiments were designed to demonstrate the possible inadequacy of the dual spectrum approach sometimes used to solve the problem of non-Poisson counting statistics encountered in loss-free counting, and to test the "variance spectrum" alternative offered by the DSPEC^{PLUS ®} . It is concluded that the DSPEC^{PLUS ®} performs with good accuracy at dead times lower than 90%, even when count rates vary. It is also concluded that the dual spectrum approach indeed is inadequate. Finally, it is shown that the "variance" spectrum approach provides the correct uncertainties to be used in the treatment of LFC or ZDT data.     

Use of near-infrared reflectance spectroscopy in predicting nitrogen,phosphorus and calcium contents in heterogeneous woody plant species

Near-infrared reflectance spectroscopy was applied to determine nitrogen (N), phosphorus (P) and calcium (Ca) content in leaf samples of 18 woody species. A total of 183 samples from mountain, riparian and dry areas from the Central–Western Iberian Peninsula were collected for this purpose. The wide intervals of variation observed in nutrient concentrations (6.6–45.0 g kg^–1 for N, 0.24–2.97 g kg^–1 for P, and 1.00–20.06 g kg^–1 for Ca) were due to the great heterogeneity of the samples. To develop calibration equations, multiple linear regression, and partial least-squares regression (PLSR) were used. In both cases, three mathematical transformations of the data were applied: log1/R and first and second derivatives. The best calibration statistics were obtained using PLSR and derivative transformations (second derivative for N and first derivative for P and Ca). The following coefficients of multiple determination (R²) and standard errors of cross validation were obtained: 0.99 and 0.93 for N, 0.94 and 0.15 for P, and 0.95 and 0.88 for Ca. In the external validation the standard errors of prediction obtained were 0.76 (N), 0.11 (P) and 0.60 (Ca).     

Comparison of three different enrichment strategies for serum low molecular weight protein identification using shotgun proteomics approach

Serum low-molecular weight (LMW) proteins potentially contain useful biological information and their identification can be used to discover novel potential biomarkers. Given the high complexity of serum samples, in the last years several different prefractionation and enrichment strategies have been developed. In this study three different methods, i.e. hydrogel nanoparticles, Proteominer^® peptide ligand affinity beads and Sartorius Vivaspin^® centrifugal ultrafiltration device, were compared and evaluated in order to select the best strategy for the enrichment and prefractionation of LMW proteins. A shotgun proteomics approach was adopted, with in-solution proteolytic digestion of the whole protein mixture and determination of the resulting peptides by nanoHPLC coupled with a high-resolution Orbitrap LTQ-XL mass spectrometer. Data analysis, focusing on the LMW proteome (MW ≤ 40 kDa), has shown that the hydrogel nanoparticles performed better in enriching the LMW protein profiles, with 115 proteins identified against 93 and 95 for Proteominer^® beads and Sartorius Vivaspin^® device, respectively.     

Carbon nuclear magnetic resonance spectroscopic fingerprinting of commercial gasoline: Pattern-recognition analyses for screening quality control purposes

In this work, the combination of carbon nuclear magnetic resonance (¹³C NMR) fingerprinting with pattern-recognition analyses provides an original and alternative approach to screening commercial gasoline quality. Soft Independent Modelling of Class Analogy (SIMCA) was performed on spectroscopic fingerprints to classify representative commercial gasoline samples, which were selected by Hierarchical Cluster Analyses (HCA) over several months in retails services of gas stations, into previously quality-defined classes. Following optimized ¹³C NMR-SIMCA algorithm, sensitivity values were obtained in the training set (99.0%), with leave-one-out cross-validation, and external prediction set (92.0%). Governmental laboratories could employ this method as a rapid screening analysis to discourage adulteration practices.     

Computational modelling of the antischistosomal activity for neolignan derivatives based on the MIA-SAR approach

Theoretical models for exploring the antischistosomal activity of a dataset of 18 synthetic neolignans are built using the multivariate image analysis applied to structure–activity relationships (MIA-SAR) approach. The obtained models were validated using the accuracy (Acc) in leave-one-out cross-validation, external validation and Y-randomization procedures, yielding correct classification superior to 80%, 70% and 60%, respectively. Additionally, a comparison was made of the models obtained from binary (black and white) and coloured images; the colours (pixel values) were selected to correspond to chemical properties. It was observed that the models obtained from coloured images with pixel values corresponding to electronegativity (known as the aug-MIA-SAR_colour approach) generally yielded superior statistical parameters compared with those obtained from binary images (MIA-SAR) and randomly coloured images (atoms are coloured according to their type) with atomic sizes corresponding to Van der Waals radius (aug-MIA-SAR), respectively. Mechanistic interpretation of the influence of different substituents on the antischistosomal activity revealed that methoxy substituents in the R1 (or R2) and R5 positions of the neolignan scaffold are indispensable for the antischistosomal activity. The obtained results provide knowledge of the possible structural modifications to yield novel neolignan compounds with antischistosomal activity.     

FT-midIR determination of fatty acid profiles, including trans fatty acids, in bakery products after focused microwave-assisted Soxhlet extraction

A study of the feasibility of Fourier transform medium infrared spectroscopy (FT-midIR) for analytical determination of fatty acid profiles, including trans fatty acids, is presented. The training and validation sets—75% (102 samples) and 25% (36 samples) of the samples once the spectral outliers have been removed—to develop FT-midIR general equations, were built with samples from 140 commercial and home-made bakery products. The concentration of the analytes in the samples used for this study is within the typical range found in these kinds of products. Both sets were independent; thus, the validation set was only used for testing the equations. The criterion used for the selection of the validation set was samples with the highest number of neighbours and the most separation between them (H<0.6). Partial least squares regression and cross validation were used for multivariate calibration. The FT-midIR method does not require post-extraction manipulation and gives information about the fatty acid profile in two min. The 14:0, 16:0, 18:0, 18:1 and 18:2 fatty acids can be determined with excellent precision and other fatty acids with good precision according to the Shenk criteria, R ²≥0.90, SEP=1–1.5 SEL and R ²=0.70-0.89, SEP=2–3 SEL, respectively. The results obtained with the proposed method were compared with those provided by the conventional method based on GC-MS. At 95% significance level, the differences between the values obtained for the different fatty acids were within the experimental error. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

HPLC fluorescence determination of nitrites in water using precolumn derivatization with 4-methyl-7-aminocoumarin

Summary A rapid, simple, and sensitive method is described for determination of nitrites in water. Nitrite (NO₂–) ions react with coumarin 120^® (4-methyl-7-aminocoumarin) in sulfuric acid medium to give the corresponding 7-diazo compound. After hydrolysis, this latter yields (95%) the highly fluorescent 4-methyl-7-hydroxycoumarin (4-methylumbelliferone) which is fluorimetrically detected at 380 nm after excitation at 325 nm.In order to avoid interference from both excess coumarin 120^® and the trace amounts of 4-methylumbelliferone which occurs in coumarin 120^® as an impurity, use of HPLC is mandatory; a satisfactory separation is obtained on a cyano stationary phase with apolar hexane-isopropanol (955, v/v) as eluent. Under these conditions, linearity of response is obtained from 1 to 30 g.L^–1 of NO₂–; the limit of detection is 0.5 g.L^–1. The repeatability and reproducibility, expressed as RSD %, are 2.5 and 4.7 % respectively, for n=6 and 5 g.L^–, analytical characteristics which demonstrate the reliability of the proposed method.     

Physicochemical characterization of ion-exchange membranes in water-methanol mixtures

A complete physicochemical characterization of two ion-exchange membranes—CM2 and Nafion^®117—used in electrodialysis and in direct methanol fuel cells (DMFC) has been carried out. For each membrane, in different methanol-water mixtures—0%, 20%, 40%, 60%, 80% and 100%—and at different temperatures (25.0; 40.0 et 55.0 °C), we have measured the variations of the geometrical dimensions, the proton electrical conductivity, the swelling rate and the amount of methanol in the membrane. The FTIR analysis of Nafion^®117 was performed at different methanol contents of the external solution.The results show that the CM2 membrane presents the best geometrical stability, and the lowest conductivity at any methanol content. At high methanol contents, Nafion^®117 is 10 times more conductive than the CM2 membrane. It was found that the methanol is absorbed more by Nafion^®117, and its effect is more noticeable on the microstructure of this membrane, under standard conditions. The high methanol permeability of these membranes, particularly of the Nafion^®117, induces bad cell efficiencies and lifetimes.     

Identification of toxic compounds in wastewater treatment plants during a field experiment

Chemical analysis and toxicity bioassays were used in conjunction to determine the toxic compounds present in wastewater. This combined methodology was applied to wastewater samples collected at two wastewater treatment plants (WWTP) from the area of Barcelona (Spain), during a field experiment carried out from 1-4 April 2000. The efficiency of the WWTP was evaluated by collecting and analyzing samples at various stages of the water treatment process. The samples corresponded to the raw influent, from first settlement—before biological treatment—and from the effluent.Two bioluminescence inhibition assays: ToxAlert^®10 and ToxAlert^®100 from Merck both based on the bioluminescence inhibition of Vibrio fischeri (marine bacterial specie) were used. ToxAlert^®10 is a portable device with no temperature control and uses freeze-dried bacterial reagent and ToxAlert^®100 uses liquid-dried bacterial reagent and the incubation takes place at controlled temperature. Both tests showed similar results.Besides the toxicity studies, the wastewater samples were characterized by various analytical protocols involving the use of solid phase extraction (SPE) followed by liquid chromatography-mass spectrometry (LC-MS). Phenols, non-ionic surfactants, linear alkyl benzene sulphonates, benzene and naphthalene sulphonates and micro-pollutants with high endocrine-disrupting effects like estradiol and ethynyl estradiol were identified at the WWTP.The toxic responses obtained for the samples collected at WWTP were defined by the 50% effective concentration (EC₅₀), the Toxicity Units (TU) and the toxicity impact index (TII₅₀). The toxic effect at the different steps of the WWTP was attributed to the compounds identified and quantified by LC-MS like transformation products of nonylphenol polyethoxylate such as nonylphenol and nonylphenol carboxylate.     

Nuclear fluorination of 2,4-diarylthiazoles with Accufluor

A series of 2,4-diphenylthiazole derivatives were synthesized and directly fluorinated at the 5-position by reaction with the N-F fluorinating reagent Accufluor^®. Although fluorination occurred selectively at the thiazole ring, it was always incomplete and thus yields for the novel fluorinated products were low to moderate (19-43%) following purification to remove starting material. Nonetheless, the target compounds were obtained in a convenient and straightforward manner. Selectfluor^® was not as effective as Accufluor^® as it gave a trace amount of the 5-chlorothiazole that was difficult to remove by chromatography.     

NOR and nitroxide-based HAS in accelerated photooxidation of carbon-chain polymers; Comparison with secondary HAS: An ESRI and ATR FTIR study

Understanding processes resulting in heterogeneous degradation in polymers is of extreme importance for improving their stabilization and minimizing negative impact of photooxidation on the material properties. We adopted modern physical techniques for studies of spatial distribution of intermediates and products of photodegradation during accelerated ageing of four commodity polymers, polypropylene (PP), polyethylene (PE), polystyrene (PS) and poly(ethylene-co-norbornene) (Topas^®, TP) stabilized with hindered amine stabilizer (HAS). Concentration profiles of nitroxides inside polymer plaques along the direction perpendicular to their surface were determined by electron spin resonance imaging (ESRI) as a function of the duration of the accelerated photooxidation. We present data characterizing stabilization activity of three alkoxyamine derivatives of HAS (Tinuvin^® NOR 123, Tinuvin^® NOR 371, Flamestab^® NOR 116), Chimasorb^® 119 structurally similar to Flamestab^® NOR 116, and nitroxide-based HAS Dastib^® 1045 and compare them with the data characterizing stabilization activity of the secondary HAS (>NH) Tinuvin^® 770. ESRI data are complemented by ATR FTIR spectroscopic detection of oxidation products on the surface and inside the plaques and by data characterizing diffusive optical transmittance of the polymer plaques in the spectral region 280-400 nm (terrestrial range of the solar UV radiation).     

Miniaturized electrochemical system for cholinesterase inhibitor detection

The utility of a simple, low-cost detection platform for label-free electrochemical characterization of acetylcholinesterase (AChE) inhibition is demonstrated as a potential tool for screening of small-molecule therapeutic agents for Alzheimer's disease (AD). Technique validation was performed against the standard Ellman's colorimetric assay using the clinically established cholinesterase inhibitor (ChEI), Donepezil (Aricept^®). Electrochemical measurements were obtained by differential pulse voltammetry (DPV) performed using a portable potentiostat system for detection of the enzymatic product, thiocholine (TCh), by direct oxidation on unmodified gold screen-printed electrodes. The IC₅₀ profiles for Donepezil measured in vitro were found to be comparable between both colorimetric and electrochemical detection methods for the analysis of purified human erythrocyte-derived AChE (28 ± 7 nM by DPV; 26 ± 8 nM by Ellman's method). The selectivity of this unmodified electrode system was compared to a range of biological sulfur-containing compounds including cysteine, homocysteine, glutathione and methionine as well as ascorbic acid. Preliminary studies also demonstrated the potential applicability of this electrochemical technique for the analysis of Donepezil in crude cholinesterase samples from anterior cortex homogenates of C57BL/6J mice.     

Isolation of Pu-isotopes from environmental samples using ion chromatography for accelerator mass spectrometry and alpha spectrometry

A radiochemical method for the isolation of plutonium-isotopes from environmental samples, based on the use of specific extraction chromatography resins for actinides (TEVA^®, Eichrom Industries, Inc.), has been set up in our laboratory and optimised for their posterior determination by alpha spectrometry (AS) or accelerator mass spectrometry (AMS). The proposed radiochemical method has replaced in our lab a well-established one based on the use of a relatively un-specific anion-exchange resin (AG^® 1X8, Bio-rad Laboratories, Inc.), because it is clearly less time consuming, reduces the amounts and molarities of acid wastes produced, and reproducibly gives high radiochemical yields.In order to check the reliability of the proposed radiochemical method for the determination of plutonium-isotopes in different environmental matrixes, twin aliquots of a set of samples were prepared with TEVA^® and with AG^® 1X8 resins and measured by AS. Some samples prepared with TEVA^® resins were measured as well by AMS. As it is shown in the text, there is a comfortable agreement between AS and AMS, which adequately validates the method.