Multivariate Curve Resolution With Alternating Least Squares Research Articles

Binding Mechanism of Bovine Serum Albumin to Troxerutin by Synchronous Fluorescence Spectroscopy and Chemometrics

Previous works have reported the binding of Bovine Serum Albumin (BSA) to troxerutin by absorption and fluorescence spectroscopy. In order to extend the investigation of their binding characteristics, the study presented here was designed to evaluate their interaction by the synchronous fluorescence spectroscopy and Multivariate Curve Resolution With Alternating Least Squares (MCR-ALS) under simulative physiological conditions. The Evolving Factor Analysis (EFA) and MCR-ALS analysis results show that there are three chemical species in the troxerutin-BSA system, which reveal that the fluorescence quenching of BSA caused by troxerutin may be a static quenching mechanism. The spectral profile of each compound was obtained by the soft of MCR-ALS. The obtained average binding constant is 6.43×105 mol-1 L for BSA and troxerutin concentration ratio is 7 while that is 6.89×105 mol-1 L for troxerutin and BSA concentration ratio is 5. The study might further help us to better understand the structural features and toxicological action of troxerutin at the molecular level.

Independent components analysis as a means to have initial estimates for multivariate curve resolution-alternating least squares

Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS) is a curve resolution method based on a bilinear model which assumes that the observed spectra are a linear combination of the spectra of the pure components in the system. The algorithm steps include the determination of the number of components by rank analysis methods, initial estimates for the concentrations and/or spectra and an iterative optimization. Sometimes, suitable results may not be achieved when MCR-ALS is applied. One reason for this is the importance of the initial estimates of the spectral profiles. In that case, the MCR-ALS algorithm may reach a local minimum instead of a global minimum and this can result in ineffective curve resolution. The most popular algorithm used to find the initial estimates (PURE derived from SIMPLISMA) suffers from an essential drawback, which is the necessity to have “pure” variables related to a single spectral component, which cannot be expected in all cases because of the strong signal overlapping as in the Ultraviolet–Visible (UV–Vis) spectroscopy. This work summarizes this problem, presenting a case study based on UV–Vis spectroscopy of heated olive oil. To solve the problems of the need for “pure” variables and to avoid local minima with MCR-ALS, Independent Components Analysis (ICA) was used to calculate initial estimates for MCR-ALS. The results from this study suggest that this use of ICA prior to MCR-ALS improves the resolution for UV–Vis data and provides acceptable resolution results when compared to the most used method, PURE.

Thermally induced solid-state transformation of cimetidine. A multi-spectroscopic/chemometrics determination of the kinetics of the process and structural elucidation of one of the products as a stable N3-enamino tautomer

Exposure of cimetidine (CIM) to dry heat (160–180°C) afforded, upon cooling, a glassy solid containing new and hitherto unknown products. The kinetics of this process was studied by a second order chemometrics-assisted multi-spectroscopic approach. Proton and carbon-13 nuclear magnetic resonance (NMR), as well as ultraviolet and infrared spectroscopic data were jointly used, whereas multivariate curve resolution with alternating least squares (MCR-ALS) was employed as the chemometrics method to extract process information. It was established that drug degradation follows a first order kinetics.One of the products was structurally characterized by mono- and bi-dimensional NMR experiments. It was found to be the N3-enamino tautomer (TAU) of CIM, resulting from the thermal isomerization of the double bond of the cyanoguanidine moiety of the drug, from the imine form to its N3-enamine state.The thus generated tautomer demonstrated to be stable for months in the glassy solid and in methanolic solutions. A theoretical study of CIM and TAU revealed that the latter is less stable; however, the energy barrier for tautomer interconversion is high enough, precluding the process to proceed rapidly at room temperature.

Fourier transform infrared imaging and microscopy studies of Pinus radiata pulps regarding the simultaneous saccharification and fermentation process

The distribution and chemical patterns of lignocellulosic components at microscopic scale and their effect on the simultaneous saccharification and fermentation process (SSF) in the production of bioethanol from Pinus radiata pulps were analyzed by the application of diverse microscopical techniques, including scanning electronic microscopy (SEM), confocal laser scanning microscopy (CLSM) and attenuated total reflectance (ATR) – Fourier transform infrared microspectroscopy. This last technique was accompanied with multivariate methods, including principal component analysis (PCA) and multivariate curve resolution with alternating least squares (MCR-ALS) to evaluate the distribution patterns and to generate pure spectra of the lignocellulosic components of fibers. The results indicate that the information obtained by the techniques is complementary (ultrastructure, confocality and chemical characterization) and that the distribution of components affects the SSF yield, identifying lignin coalescence droplets as a characteristic factor to increase the SSF yield. Therefore, multivariate analysis of the infrared spectra enabled the in situ identification of the cellulose, lignin and lignin-carbohydrates arrangements. These techniques could be used to investigate the lignocellulosic components distribution and consequently their recalcitrance in many applications where minimal sample manipulation and microscale chemical information is required.

MCR OF THE QUENCHING OF THE EEM OF FLUORESCENCE OF AFLATOXINS (B1, G1) BY GOLD NANOPARTICLES

In This research, gold nanoparticles were synthesized and functionalized by the antibody of aflatoxins. The quenching of the fluorescence of excitation emission matrices (EEM) of two type of aflatoxins (B1, G1), provoked by the gold nanoparticles, was studied by principal component analysis (PCA) and multivariate curve resolution with alternating least squares (MCR-ALS). These aflatoxins show quenching in the presence of the gold-labeled antibodies. Solutions containing a constant concentration of aflatoxins and variable concentrations of gold-labeled antibodies have been prepared and the fluorescence spectra have been recorded by scanning excitation and emission wavelengths. Excitation and emission spectra and quenching profiles were extracted from EEM using MCR-ALS. Applying suitable constraints lead to satisfactory results for the determination of aflatoxins in all samples.

THERMAL DEGRADATION OF TOCOPHEROL AND OXIDATION PRODUCTS IN DIFFERENT OLIVE OIL CLASSES USING UV-VIS SPECTROSCOPY AND MCR-ALS

Extra virgin olive oil, virgin olive oil and mixed oil were analyzed by UV-Vis spectroscopy and Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS). When the oils were heated at high temperatures (frying), oxidation products were formed which were harmful to human health and antioxidants present were degraded. The aim of this study was to evaluate tocopherol (an antioxidant present in oils) and the behaviour of oxidation products in different olive oil classes. MCR-ALS results showed that degradation started at 70 oC for extra virgin olive oil, while tocopherol concentration decreased and oxidation products increased starting at 40 oC in virgin olive oil. In mixed oil, commercialized as a mixture of 85% soybean oil and 15% olive oil, tocopherol concentration started to decrease and oxidation products increase at 30 oC. The results suggested that extra virgin olive oil offered more resistance to increasing temperatures, while virgin olive oil and mixed oil were less resistant.

Detection of explosives on the surface of banknotes by Raman hyperspectral imaging and independent component analysis

The aim of this study was to develop a methodology using Raman hyperspectral imaging and chemometric methods for identification of pre- and post-blast explosive residues on banknote surfaces. The explosives studied were of military, commercial and propellant uses. After the acquisition of the hyperspectral imaging, independent component analysis (ICA) was applied to extract the pure spectra and the distribution of the corresponding image constituents. The performance of the methodology was evaluated by the explained variance and the lack of fit of the models, by comparing the ICA recovered spectra with the reference spectra using correlation coefficients and by the presence of rotational ambiguity in the ICA solutions. The methodology was applied to forensic samples to solve an automated teller machine explosion case. Independent component analysis proved to be a suitable method of resolving curves, achieving equivalent performance with the multivariate curve resolution with alternating least squares (MCR-ALS) method. At low concentrations, MCR-ALS presents some limitations, as it did not provide the correct solution. The detection limit of the methodology presented in this study was 50μgcm−2.

Monitoring simultaneous photocatalytic-ozonation of mixture of pharmaceuticals in the presence of immobilized TiO2 nanoparticles using MCR-ALS: Identification of intermediates and multi-response optimization approach

The present study has focused on the degradation of a mixture of three pharmaceuticals, i.e. methyldopa (MDP), nalidixic acid (NAD) and famotidine (FAM) which were quantified simultaneously during photocatalytic-ozonation process. The experiments were conducted in a semi-batch reactor where TiO2 nanoparticles (crystallites mean size 8nm) were immobilized on ceramic plates irradiated by UV-A light in the proximity of oxygen and/or ozone. The surface morphology and roughness of the bare and TiO2-coated ceramic plates were analyzed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). An analytical methodology was successfully developed based on both recording ultraviolet–visible (UV–Vis) spectra during the degradation process and a data analysis using multivariate curve resolution with alternating least squares (MCR-ALS). This methodology enabled the researchers to obtain the concentration and spectral profiles of the chemical compounds which were involved in the process. A central composite design was used to study the effect of several factors on multiple responses namely MDP removal (Y1), NAD removal (Y2) and FAM removal (Y3) in the simultaneous photocatalytic-ozonation of these pharmaceuticals. A multi-response optimization procedure based on global desirability of the factors was used to simultaneously maximize Y1, Y2 and Y3. The results of the global desirability revealed that 8mg/L MAD, 8mg/L NAD, 8mg/L FAM, 6L/h ozone flow rate and a 30min-reaction time were the best conditions under which the optimized values of various responses were Y1=95.03%, Y2=84.93% and Y3=99.15%. Also, the intermediate products of pharmaceuticals generated in the photocatalytic-ozonation process were identified by gas chromatography coupled to mass spectrometry.

Parallel factor analysis and multivariate curve resolution as data fusion tools to supervise a stream

In this work, a new method is proposed to monitor the distribution, evolution and correlation of dissolved organic matter on the superficial water of a stream with respect to physicochemical variables that characterize the basin and season sampling of each campaign. The method is based on measuring fluorescence emission–excitation matrices and some physicochemical parameters of water samples through both time and space. In a first phase, parallel factor analysis (PARAFAC) or multivariate curve resolution with alternating least-squares (MCR-ALS) were applied to extract the information on the relative proportions of each fluorophore on each sample. Then, MCR-ALS was applied again to the entire database, in order to study the spatial and time distribution. This methodology was used to study the behavior of a basin stream that is significantly modified by anthropic activities.

Liquid chromatography with diode array detection and multivariate curve resolution for the selective and sensitive quantification of estrogens in natural waters

Following the green analytical chemistry principles, an efficient strategy involving second-order data provided by liquid chromatography (LC) with diode array detection (DAD) was applied for the simultaneous determination of estriol, 17β-estradiol, 17α-ethinylestradiol and estrone in natural water samples. After a simple pre-concentration step, LC–DAD matrix data were rapidly obtained (in less than 5min) with a chromatographic system operating isocratically. Applying a second-order calibration algorithm based on multivariate curve resolution with alternating least-squares (MCR-ALS), successful resolution was achieved in the presence of sample constituents that strongly coelute with the analytes. The flexibility of this multivariate model allowed the quantification of the four estrogens in tap, mineral, underground and river water samples. Limits of detection in the range between 3 and 13ngL−1, and relative prediction errors from 2 to 11% were achieved.

Thermal edible oil evaluation by UV–Vis spectroscopy and chemometrics

Edible oils such as colza, corn, sunflower, soybean and olive were analysed by UV–Vis spectroscopy and Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS). When vegetable oils were heated at high temperatures (frying), oxidation products were formed which were harmful to human health in addition to degrading the antioxidants present, and this study aimed to evaluate tocopherol (one antioxidant present in oils) and the behaviour of oxidation products in edible oils. The MCR-ALS results showed that the degradation started at 110°C and 85°C, respectively, for sunflower and colza oils, while tocopherol concentration decreased and oxidation products increased starting at 70°C in olive oil. In soybean and corn oils, tocopherol concentration started to decrease and oxidation products increased at 50°C. The results suggested that sunflower, colza and olive oils offered more resistance to increasing temperatures, while soybean and corn oils were less resistant.

A dynamic thermal ATR-FTIR/chemometric approach to the analysis of polymorphic interconversions. Cimetidine as a model drug

Crystal polymorphism of active ingredients is relevant to the pharmaceutical industry, since polymorphic changes taking place during manufacture or storage of pharmaceutical formulations can affect critical properties of the products. Cimetidine (CIM) has several relevant solid state forms, including four polymorphs (A, B, C and D), an amorphous form (AM) and a monohydrate (M1). Dehydration of M1 has been reported to yield mixtures of polymorphs A, B and C or just a single form.Standards of the solid forms of CIM were prepared and unequivocally characterized by FTIR spectroscopy, digital microscopy, differential scanning calorimetry and solid state 13C NMR spectroscopy. Multivariate curve resolution with alternating least squares (MCR-ALS) was coupled to variable temperature attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) to dynamically characterize the behavior of form M1 of CIM over a temperature range from ambient to 160°C, without sample pretreatment.MCR-ALS analysis of ATR-FTIR spectra obtained from the tested solid under variable temperature conditions unveiled the pure spectra of the species involved in the polymorphic transitions. This allowed the simultaneous observation of thermochemical and thermophysical events associated to the changes involved in the solid forms, enabling their unequivocal identification and improving the understanding of their thermal behavior.It was demonstrated that under the experimental conditions, dehydration of M1 initially results in the formation of polymorph B; after melting and upon cooling, the latter yields an amorphous solid (AM). It was concluded that the ATR-FTIR/MCR association is a promising and useful technique for monitoring solid-state phase transformations.

Chemometric modeling of organic contaminant sources in surface waters of a mediterranean river basin

Chemometric methods are applied to the analysis and interpretation of large multivariate datasets obtained in environmental monitoring studies. Concentrations of multiple organic compounds were measured in river samples taken from several sampling sites, at various geographical locations, during a number of campaigns and/or sampling time periods. Samples were collected and analyzed as part of an extensive multi-annual monitoring program from a mediterranean river basin (in Catalonia, at the northeast of Spain) by the Water Quality Regional Agency. Due to the great amount of multivariate data stored in environmental databases and to their complexity, chemometric modeling methods such as Principal Component Analysis (PCA) and Multivariate Curve Resolution with Alternating Least-Squares (MCR-ALS) coupled with appropriate mapping representations are proposed for the evaluation of the environmental quality of the studied rivers. Results achieved in this study are intended to be a contribution to water quality assessment and evaluation of contamination of surface waters in river basins, and to support public policies of environmental control and management of the regions under study.

MULTIVARIATE CURVE RESOLUTION WITH ALTERNATING LEAST SQUARES: DESCRIPTION, OPERATION AND APLICATIONS

Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS) is a resolution method that has been efficiently applied in many different fields, such as process analysis, environmental data and, more recently, hyperspectral image analysis. When applied to second order data (or to three-way data) arrays, recovery of the underlying basis vectors in both measurement orders (i.e. signal and concentration orders) from the data matrix can be achieved without ambiguities if the trilinear model constraint is considered during the ALS optimization. This work summarizes different protocols of MCR-ALS application, presenting a case study: near-infrared image spectroscopy.

An eco-friendly strategy, using on-line monitoring and dilution coupled to a second-order chemometric method, for the construction of dissolution curves of combined pharmaceutical associations

A simple, precise, economic and minimally operator-dependent method was developed under green analytical chemistry principles, for the simultaneous construction of the dissolution curves of a pharmaceutical association in short time and without employing organic solvents, allowing important savings of labor and resources. The carvedilol (CAR) and hydrochlorothiazide (HCT) combined formulation was employed as a model. The method (OD/UV-MCR) involves on-line sample dilution (OD) and UV detection of the analytes, coupled to multivariate curve resolution with alternating least squares (MCR-ALS).OD/UV-MCR proved to be robust and was successfully validated in accordance to ICH guidelines, fulfilling acceptance criteria for specificity (r2 of spectral correlation>0.950), linearity [r>0.999 (N=25) in the ranges 1.00–31.1mgl−1 and 0.51–15.2mgl−1 for CAR and HCT, respectively] and precision (RSD<2%). Accuracy was assessed by point-to-point comparison between the dissolution profiles furnished by the proposed method with those provided by HPLC analysis, evidencing the usefulness of this monitoring system.In addition, OD/UV-MCR was successfully employed for the comparative analysis of three lots of commercial formulations of the CAR–HCT pharmaceutical association, belonging to a couple of different brands, employing Moore and Flanner's f2 similarity indicator.

A Multivariate Curve Resolution Approach to the Study of the Degradation Kinetics of Valsartan under Photolytic and Acid Conditions

ABSTRACTThe analysis of UV‐spectrophotometric data with second‐order chemometrics techniques, including multivariate curve resolution with alternating least‐squares (MCR‐ALS) and hybrid hard‐ and soft MCR (HS‐MCR), was examined as an alternative tool for studying the kinetics of drug degradation under stress conditions, employing valsartan (VAL) as a model drug. Despite small structural and spectroscopic differences between VAL and its degradation products, MCR‐ALS and HS‐MCR were able to detect the generation of two photoneutral degradation products (DP‐1 and DP‐2) and a single acid hydrolysis product (DP‐3), providing good approximations to their pure spectra and concentration profiles, from which estimations of the kinetic profiles and rate constants were obtained. Kinetic models based on first‐order reactions explained the degradation processes. MCR‐ALS and HS‐MCR analyses yielded similar rate constants; however, the latter was capable of more properly fitting the experimental data to a kinetic model in the case of drug photolysis. The results were confirmed by comparison with data obtained by HPLC analysis of the degraded samples.

Quantitative analysis of biodiesel in blends of biodiesel and conventional diesel by comprehensive two-dimensional gas chromatography and multivariate curve resolution

In this paper, a method to determine the composition of blends of biodiesel with mineral diesel (BXX) by multivariate curve resolution with Alternating Least Squares (MRC-ALS) combined to comprehensive two-dimensional gas chromatography with Flame Ionization Detection (GC×GC-FID) is presented. Chromatographic profiles of BXX blends produced with biodiesels from different sources were used as input data. An initial evaluation carried out after multiway principal component analysis (MPCA) was used to reveal regions of the chromatograms were the signal was likely to be dependent on the concentration of biodiesel, regardless its vegetable source. After this preliminary step MCR-ALS modeling was carried out only using relevant parts of the chromatograms. The resulting procedure was able to predict accurately the concentration of biodiesel in the BXX samples regardless of its origin.

Native fluorescence spectra of human cancerous and normal breast tissues analyzed with non-negative constraint methods

The native fluorescence spectra of human cancerous and normal breast tissues were investigated using the selected excitation wavelength of 340 nm to excite key building block molecules, such as reduced nicotinamide adenine dinucleotide (NADH), collagen, and flavin. The measured emission spectra were analyzed using a non-negative constraint method, namely multivariate curve resolution with alternating least-squares (MCR-ALS). The results indicate that the biochemical changes of tissue can be exposed by native fluorescence spectra analysis. The MCR-ALS-extracted components corresponding to the key fluorophores in breast tissue, such as collagen, NADH, and flavin, show differences of relative contents of fluorophores in cancerous and normal breast tissues. This research demonstrates that the native fluorescence spectroscopy measurements are effective for detecting changes of fluorophores composition in tissues due to the development of cancer. Native fluorescence spectroscopy analyzed by MCR-ALS may have the potential to be a new armamentarium.

Optical Detection of Meat Spoilage Using Fluorescence Spectroscopy with Selective Excitation Wavelength

The native fluorescence (FL) spectra of muscle foods (meat) stored at 4 °C (refrigerated) and 25 °C (at room temperature) were measured with the selected excitation wavelength of 340 nm as a function of storage time to detect the meat spoilage status. The contributions of the principal biochemical components to the FL spectra were extracted using Multivariate Curve Resolution with Alternating Least-Squares (MCR-ALS). The change of the reduced nicotinamide adenine dinucleotide (NADH) content was found from the measured FL spectra and the MCR-ALS analysis, which reflects the microbial spoilage of muscle foods involved in the metabolic processes. This study presents the possibility that the change of relative content of NADH determined by native FL spectroscopy may be used as a "fingerprint" or criterion for monitoring the spoilage status of muscle foods.

Chemometrics approach for determination and optimization of simultaneous photooxidative decolourization of a mixture of three textile dyes

In this paper simultaneous photooxidation of three textile dyes (CI Basic Red 46 (BR46), Malachite Green (MG) and CI Basic Blue 3 (BB3)) by UV/H2O2 process is reported. A rapid analytical methodology based on recording UV–Vis spectra during the photooxidative decolourization process and the data treatment using multivariate curve resolution with alternating least squares (MCR-ALS) was developed. The three textile dyes were quantified simultaneously despite the overlap of their spectra. Also, the central composite design was applied for optimization of the photooxidative decolourization of the solution containing three dyes using the UV/H2O2 process. The investigated factors (variables) were the reaction time, initial concentration of the three dyes and initial concentration of H2O2. The optimum initial concentration of the three dyes, H2O2 initial concentration and reaction time were found to be 4 mg/L, 48 mg/L and 30 min, respectively. The photodegradation products of three dyes were identified by the GC–MS technique.