Role of the co-surfactant nature in soybean w/o microemulsions.

Abstract The influence of the co-surfactant on physicochemical properties of w/o soybean oil microemulsions (MEs) has been studied. In spite of the similarity in phase diagrams, the MEs display remarkable differences when examined by electrical conductivity, dynamic light scattering (DLS), small angle X-ray scattering (SAXS) and linear voltammetry. When different short-chain alcohols were employed as co-surfactants, together with sodium dodecyl sulfate (SDS) as surfactant, the DLS results indicated the systems to be monodisperse. Both the electrical conductivity of the MEs and the hydrodynamic radii of the droplets (RH) increased with water content while RH diminished as temperature increas…

Use of linear discriminant analysis applied to vibrational spectroscopy data to characterize commercial varnishes employed for art purposes.

An improvement of methodologies for characterising synthetic resins used in varnishes employed for art purposes has been suggested. Several kinds of standard of the most common polymeric resins (acrylic, vinyl, poly(vinyl alcohol), alkyd, cellulose nitrate, latex, polyester, polyurethane, epoxy, organosilicic, and ketonic) were analyzed by Fourier transform infrared (FTIR) spectroscopy. Synthetic resins characterization is based on the mathematical treatment of their whole spectrum, dividing it in 13 sections, avoiding the one-by-one interpretation of the absorption bands. The mathematical model takes as variables the maximal absorbance of each section, and each synthetic standard resin as …

Chemical initiation for butyl and lauryl acrylate monolithic columns for CEC.

Butyl acrylate (BA)- and lauryl acrylate (LA)-based monolithic stationary phases for CEC were synthesized, using a redox system as initiator of polymerization. BA monoliths were initiated with ammonium peroxodisulfate, whereas LA columns were obtained with lauroyl peroxide as initiator. In both cases, TEMED was used to activate the process. The influence of porogenic solvent composition on both morphological and electrochromatographic properties of the resulting monoliths was investigated. Excellent efficiencies (minimum plate heights of 4.2-6.3 microm for BA columns and 2.6-5.3 microm for LA stationary phases, for a PAHs mixture) were achieved. The capability of separation of both types of…

Lauroyl peroxide as thermal initiator of lauryl methacrylate monolithic columns for CEC

The preparation of lauryl methacrylate (LMA)-based monolithic columns for CEC using lauroyl peroxide (LPO) as thermal initiator of polymerization has been investigated. The influence of initiator amount and composition of porogenic solvent on the physical and electrochromatographic properties of the resulting LMA-based monoliths was evaluated. A comparison with LMA-based columns thermally polymerized with AIBN was performed. At a given porogenic solvent composition, LMA stationary phases initiated with LPO showed higher permeabilities and better efficiency values than those prepared using AIBN as initiator. The optimum polymerization mixture found for LPO initiator provided a minimum plate …

Use of electronic nose to determine defect percentage in oils. Comparison with sensory panel results

Abstract An electronic nose based on an array of 6 metal oxide semiconductor sensors was used, jointly with linear discriminant analysis (LDA) and artificial neural network (ANN) method, to classify oils containing the five typical virgin olive oil (VOO) sensory defects (fusty, mouldy, muddy, rancid and winey). For this purpose, these defects, available as single standards of the International Olive Council, were added to refined sunflower oil. According to the LDA models and the ANN method, the defected samples were correctly classified. On the other hand, the electronic nose data was used to predict the defect percentage added to sunflower oil using multiple linear regression models. All …

Classification of vegetable oils according to their botanical origin using amino acid profiles established by direct infusion mass spectrometry.

Amino acid profiles, established by direct infusion mass spectrometry, have been used to classify vegetable oils according to their botanical origin. The proteins present in hazelnut, sunflower, corn, soybean, olive, avocado, peanut and grapeseed oils were precipitated with acetone, and the residue was hydrolyzed in acid medium, diluted in a hydrochloric acid/ethanol mixture, and infused into the mass spectrometer. The spectra of the hydrolyzed protein extracts showed [M+H]+ ions of the following amino acids: glycine, alanine, serine, proline, valine, threonine, cysteine, isoleucine + leucine, aspartic acid, lysine, glutamic acid, methionine, histidine, phenylalanine, arginine and tyrosine.…

Bio-metal-organic frameworks for molecular recognition and sorbent extraction of hydrophilic vitamins followed by their determination using HPLC-UV

A bio-metal-organic framework (bio-MOF) derived from the amino acid L-serine has been prepared in bulk form and evaluated as sorbent for the molecular recognition and extraction of B-vitamins. The functional pores of bio-MOF exhibit high amounts of hydroxyl groups jointly directing other supramolecular host-guest interactions thus providing the recognition of B-vitamins in fruit juices and energy drinks. Single-crystal X-ray diffraction studies reveal the specific B-vitamin binding sites and the existence of multiple hydrogen bonds between these target molecules and the framework. It offered unique snapshots to accomplish an efficient capture of these solutes in complex aqueous matrices. Fo…

Authentication of extra virgin olive oils by Fourier-transform infrared spectroscopy

Fourier-transform infrared spectroscopy (FTIR), followed by multivariate treatment of the spectral data, was used to classify vegetable oils according to their botanical origin, and also to establish the composition of binary mixtures of extra virgin olive oil (EVOO) with other low cost edible oils. Oil samples corresponding to five different botanical origins (EVOO, sunflower, corn, soybean and hazelnut) were used. The wavelength scale of the FTIR spectra of the oils was divided in 26 regions. The normalized absorbance peak areas within these regions were used as predictors. Classification of the oil samples according to their botanical origin was achieved by linear discriminant analysis (…

Orange proteomic fingerprinting: From fruit to commercial juices.

Combinatorial peptide ligand library technology, coupled to mass spectrometry, has been applied to extensively map the proteome of orange pulp and peel and, via this fingerprinting, to detect its presence in commercial orange juices and drinks. The native and denaturing extraction protocols have captured 1109 orange proteins, as identified by LC-MS/MS. This proteomic map has been searched in an orange concentrate, from a Spanish juice manufacturer, as well as in commercial orange juices and soft drinks. The presence of numerous orange proteins in commercial juices has demonstrated the genuineness of these products, prepared by using orange fruits as original ingredients. However, the low nu…

Methacrylate ester-based monolithic columns for nano-LC separation of tocopherols in vegetable oils

The separation and determination of tocopherols (Ts) in vegetable oils by nano-LC chromatography with UV-vis detection using lauryl methacrylate ester-based monolithic columns has been developed. The separation of Ts was optimized in terms of mobile phase composition on the basis of the best compromise among efficiency, resolution and analysis time. Using a mobile phase composed of ACN/methanol/water, an excellent resolution between Ts was achieved within 18 min. The LODs were lower than 0.26 μg/mL, being repeatability values of retention time and peak area below 0.15 and 3.1%, respectively. The method was applied to the quantification of Ts and tocotrienols present in several vegetable oil…

Capillary electrophoresis enhanced by automatic two-way background correction using cubic smoothing splines and multivariate data analysis applied to the characterisation of mixtures of surfactants

Mixtures of the surfactant classes coconut diethanolamide, cocamido propyl betaine and alkylbenzene sulfonate were separated by capillary electrophoresis in several media containing organic solvents and anionic solvophobic agents. Good resolution between both the surfactant classes and the homologues within the classes was achieved in a BGE containing 80 mM borate buffer of pH 8.5, 20% n-propanol and 40 mM sodium deoxycholate. Full resolution, assistance in peak assignment to the classes (including the recognition of solutes not belonging to the classes), and improvement of the signal-to-noise ratio was achieved by multivariate data analysis of the time-wavelength electropherograms. Cubic s…

Recognition and alignment of variables from UV–vis chromatograms and application to industrial enzyme digests classification

Abstract In the last years, industrial applications of chemometrics have largely increased due to their capacity to extract important information from complex records as chromatograms or spectra data. The use of chemometric methods also can avoid the use of detectors of elevated cost. In this work, a procedure to recognize the relevant chemical information contained in complex UV–vis chromatograms, after a trypsin digestion, to identify the three enzyme main classes (proteases, amylases and cellulases) commonly employed in the cleaning industry, has been developed. In order to recognize the chromatogram peaks, six indices of peak identity or identifiers were defined. A program written in MA…

Determination of cow’s milk in non-bovine and mixed cheeses by capillary electrophoresis of whey proteins in acidic isoelectric buffers

An improved method for the determination of cow's milk in non-bovine cheese is reported: electrophoresis of whey proteins in acidic, isoelectric buffers. Two background electrolytes (BGEs) have been tested: (i) 50 mM iminodiacetic acid (pH=isoelectric point=2.30 at 25 degrees C), 0.5% hydroxyethylcellulose, 0.1% Tween 20 and 6 M urea (apparent pH 3.1), E=300 V/cm, for the separation of alpha-lactalbumins (alpha-LAs); (ii) a BGE with the same composition, but supplemented with 10% Tween 20, E=450 V/cm, for the fractionation of beta-lactoglobulins (beta-LGs). Surfactants have a discriminating effect on the retention behaviour of the bovine alpha-LA and beta-LG proteins, owing to the different…

Separation and determination of homologues of linear alkylbenzenesulfonates by nonaqueous capillary zone electrophoresis using alkylammonium salts in ethanol

The separation of linear alkylbenzene sulfonates (LAS) by nonaqueous capillary electrophoresis (NACE) using negative polarity, and a buffer containing acetic acid and an alkylamine in nonaqueous ethanol, has been investigated. Several primary, secondary, and tertiary alkylamines with alkyl chains of different length were compared. The solutes travelled against the electroosmotic flow (EOF), and at the same time were braked by association with the alkylamine molecules or with the alkylammonium ions. The best resolution between adjacent LAS homologues (R approximately 2.1), partial isomer resolution in two peaks, and at the same time an excellent repeatability, was obtained with a small dipen…

Prediction of the curing time of Spanish hams using peptide profiles established by capillary zone electrophoresis

Abstract A capillary zone electrophoresis procedure for the prediction of curing time of Spanish hams using peptide profiles has been developed. Excellent resolution between the seven peptide peaks was achieved within 30 min analysis time with a BGE containing 60 mM MgSO 4 and 50 mM phosphate at pH 2.5. Using hams with curing times of 6, 8 and 12 months, both linear discriminant analysis (LDA) and multiple linear regression (MLR) models were constructed. In both cases, two different normalisation procedures of the peak areas were compared. Using LDA, all the ham samples corresponding to the three categories were correctly classified. Using MLR, the ham curing time could be predicted with av…

Determination of non-ionic and anionic surfactants in industrial products by separation on a weak ion-exchanger, derivatization and liquid chromatography

Abstract A method for the determination of priority surfactants, including fatty alcohol ethoxylates (FAE), alkylether sulfates (AES) and linear alkylbenzene sulfonates (LAS) is described. The samples were diluted with 50% methanol at pH 4 prior to solid-phase extraction on a weak anionic exchanger (WAX). The AES and LAS surfactant classes were retained, whereas the non-ionic components, including most FAE oligomers were eluted. After washing the WAX cartridge to remove cations, the remaining hydrophobic FAE oligomers were eluted using hot 80% methanol at pH 4 (at ca. 50 °C). These two eluates were combined to constitute the non-ionic fraction. Then, AES and LAS were eluted using 80% MeOH w…

Influence of photo-initiators in the preparation of methacrylate monoliths into poly(ethylene-co-tetrafluoroethylene) tubing for microbore HPLC.

[EN] In this study, poly(butyl methacrylate-co-ethyleneglycol dimethacrylate) polymeric monoliths were in situ developed within 0.75 mm i.d. poly(ethylene-co-tetrafluoroethylene) (ETFE) tubing by UV polymerization via three different free-radical initiators fscce-azobisisobutyronitrile (AIBN), 2,2-dimethoxy-2-phenylacetophenone (DMPA) and 2-methyl-4'-(methylthio)-2-morpholinopropiophenone (MTMPP). The influence of the nature of each photo-initiator and irradiation time on the morphological features of the polymer was investigated by scanning electron microscopy, and the chromatographic properties of the resulting microbore columns were evaluated using alkyl benzenes as test substances. The …

Determination of cow's milk and ripening time in nonbovine cheese by capillary electrophoresis of the ethanol-water protein fraction

A novel method is reported for analyzing adulteration of goat and ewe cheeses with cow's milk: capillary zone electrophoresis (CZE) in isoelectric, acidic buffers (50 mM imino diacetic acid, IDA, pH = pI 2.3). The cheese samples were extracted with a 20:80 v/v ethanol-water mixture in presence of 3 M urea and 1% beta-mercaptoethanol for 1 h. After centrifugation and lipid extraction, the samples were dissolved in 50 mM IDA, 6 M urea and 0.5% hydroxyethyl cellulose and analyzed by CZE at 700 V/cm. A total of 18 characteristic peaks were resolved among the three types of cheeses and 18 variables were defined as their respective areas. There was excellent similarity among the electrophoretic p…

Prediction of the genetic variety of Spanish extra virgin olive oils using fatty acid and phenolic compound profiles established by direct infusion mass spectrometry.

Abstract The genetic varieties of Spanish extra virgin olive oils (Arbequina, Hojiblanca and Picual) were predicted by direct infusion of the samples in the electrospray ionization source of a mass spectrometer, followed by linear discriminant analysis of the spectral data. The samples were 1:50 diluted (v/v) with an 85:15 propanol/methanol (v/v) mixture containing 40 mM KOH and infused. The abundances of the [M–H] − peaks of the free fatty acids (7 peaks) and 28 phenolic compounds (20 peaks) were measured. Ratios of pairs of peak abundances were used as predictors in the construction of the linear discriminant analysis models. An excellent resolution between the three genetic varieties was…

Characterization of hydroxyaromatic compounds in vegetable oils by capillary electrophoresis with direct injection in an oil-miscible KOH/propanol/methanol medium.

The separation of hydroxyaromatic compounds in vegetable oils, including synthetic antioxidants (3-tert-butyl-4-hydroxyanisol and 2,6-di-tert-butyl-4-hydroxytoluene), E-vitamers and other natural oil components, by nonaqueous capillary electrophoresis in an oil-miscible background electrolyte (BGE) was investigated. The BGE contained 40 mM KOH in a methanol/1-propanol (PrOH) mixture (15:85 v/v). The oil samples were 1:1 diluted with PrOH and directly injected in the capillary. Under negative polarity (cathode at the injection end), the anionic solutes moved faster than the electroosmotic flow, being well-resolved among them and from the triacylglycerols. Using virgin palm, extra virgin oliv…

Chemical analysis and antioxidant activity of the essential oils of three Piperaceae species growing in the central region of Cuba.

The present study describes the phytochemical profile and antioxidant activity of the essential oils of three Piperaceae species collected in the central region of Cuba. The essential oils of Piper aduncum, P. auritum and P. umbellatum leaves, obtained by hydrodistillation, were analyzed by gas chromatography-mass spectrometry. The main components of P. aduncum oil were piperitone (34%), camphor (17.1%), camphene (10.9%), 1,8-cineol (8.7%) and viridiflorol (7.4%), whereas that of P. auritum and P. umbellatum was safrole (71.8 and 26.4%, respectively). The antioxidant properties of the essential oils were also evaluated using several assays for radical scavenging ability (DPPH test and redu…

Enzyme class identification in cleaning products by hydrolysis followed by derivatization with o-phthaldialdehyde, HPLC and linear discriminant analysis.

The enzymes present in raw materials of the cleaning industry (enzyme industrial concentrates) and in household cleaners were isolated by precipitation with acetone and hydrolyzed with HCl. The resulting amino acids were derivatized with o-phthaldialdehyde, and the derivatives were separated by HPLC. The peaks of 14 amino acids were observed using a C18 column and a multi-segmented gradient of acetonitrile-water in the presence of a 5 mM citric/citrate buffer of pH 6.5. Using either normalized peak areas (divided by the sum of the peak areas of the chromatogram) or ratios of pairs of peak areas as predictor variables, linear discriminant analysis models, capable of predicting the enzyme cla…

Rapid differentiation of commercial juices and blends by using sugar profiles obtained by capillary zone electrophoresis with indirect UV detection.

A method for the determination of sugars in several fruit juices and nectars by capillary zone electrophoresis with indirect UV-vis detection has been developed. Under optimal conditions, commercial fruit juices and nectars from several fruits were analyzed, and the sugar and cyclamate contents were quantified in less than 6 min. A study for the detection of blends of high-value juices (orange and pineapple) with cheaper alternatives was also developed. For this purpose, different chemometric techniques, based on sugar content ratios, were applied. Linear discriminant analysis showed that fruit juices can be distinguished according to the fruit type, juice blends also being differentiated. …

Efficient Extraction of Olive Pulp and Stone Proteins by using an Enzyme-Assisted Method

An efficient protein extraction protocol for proteins from olive pulp and stone by using enzymes was developed. For this purpose, different parameters that affect the extraction process, such as enzyme type and content, pH, and extraction temperature and time, were tested. The influence of these factors on protein recovery was examined using the standard Bradford assay, while the extracted proteins were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The best extraction conditions were achieved at pH 7.0 and 5% (v/v) Palatase® 20000 L (lipase) for pulp and Lecitase® Ultra (phospholipase) for stone proteins. The optimal extraction temperature and time w…

Host-guest interactions for extracting antibiotics with a γ-cyclodextrin poly(glycidyl-co-ethylene dimethacrylate) hybrid sorbent

Abstract A procedure for the solid-phase extraction of antibiotics (enoxacin, ofloxacin, norfloxacin, ciprofloxacin, and sparfloxacin) in water has been developed. The sorbent used is based on a poly(glycidyl-co-ethylene dimethacrylate) network, whose previously modified surface has been functionalized with γ-cyclodextrin through a click-chemistry reaction. The architecture of the material has been characterized by thermogravimetric analysis, N2 adsorption-desorption, Raman spectroscopy, confocal microscopy, and scanning electron microscopy, showing good capability to be used as a filler for extraction cartridges. The optimization of the extraction methodology shows good intra-day and inter…

Comparison on photo-initiators for the preparation of methacrylate monolithic columns for capillary electrochromatography.

The synthesis of lauryl methacrylate monoliths for capillary electrochromatography by UV polymerization using several free-radical initiators (alpha,alpha'-azobisisobutyronitrile, 2,2-dimethoxy-2-phenylacetophenone, dibenzoyl peroxide (BPO) and lauroyl peroxide (LPO)) has been investigated. Using a 1,4-butanediol/1-propanol mixture as porogenic solvent, the influence of each initiator and its content on the morphological and electrochromatographical properties of beds was evaluated. Under their respective optimum content, satisfactory separations of a test mixture of PAHs with similar efficiencies (minimum plate heights of 8.0-12.7 microm obtained from Van Deemter plots) were achieved for t…

Determination of thyreostatics in animal feed by micellar electrokinetic chromatography

The determination of the thyreostatics 2-thiouracil, its derivatives (4-methyl-2-thiouracil, 4-propyl-2-thiouracil and 4-phenyl-2-thiouracil) and methimazole in manufactured dried animal feed by micellar electrokinetic chromatography (MEKC) is described. A 99 +/- 5% extraction yield at the 20 micrograms g-1 level (n = 8) was achieved by shaking the milled fodder with methanol-1 M NaOH (80 + 20). Aliquots of the supernatant were injected in a 75 microns x 33.5 cm uncoated silica capillary using pressure; separation was performed at 23 degrees C with 15 kV (positive polarity) in a background electrolyte (BGE) containing 40 mM sodium dihydrogenphosphate, 50 mM sodium dodecyl sulfate and 15 mM …

Preparation and evaluation of butyl acrylate-based monolithic columns for CEC using ammonium peroxodisulfate as a chemical initiator.

Acrylate-ester-based monoliths for CEC using peroxodisulfate as a chemical initiator were prepared. The influence of two ternary porogenic solvents on the physical and chromatographic properties of butyl acrylate monolithic stationary phases was investigated. The composition and the ratio of porogenic solvent were adjusted to obtain highly permeable rigid monoliths with adequate column efficiency. Among the prepared butyl acrylate monoliths, those polymerized from a ternary porogenic solvent of acetonitrile/ethanol/water exhibited the most promising performance with a minimum plate height for naphthalene of 10.5 microm and a bed permeability of 7.3 x 10(-14) m(2). A comparison in terms of e…

Metal oxide semiconductor sensors for monitoring of oxidative status evolution and sensory analysis of virgin olive oils with different phenolic content

An electronic nose based on an array of six metal oxide semiconductor sensors was used to monitor the oxidative status of virgin olive oils (VOO) during storage. VOO samples, with and without phenolic compounds, were stored at 60 °C for 7 weeks. Once a week, absorbance at 232 and 270 nm, oxidized stability index, electronic nose, and sensory analysis were evaluated. Linear discriminant analysis models were constructed in order to classify samples according to oxidative levels. Based on these models, VOO samples with and without phenolic compounds at different storage times, divided in eight categories, were correctly classified also achieving a good correlation for sensory analysis. The met…

Photo-polymerized lauryl methacrylate monolithic columns for CEC using lauroyl peroxide as initiator

Lauryl methacrylate (LMA)-ester based monolithic columns photo-polymerized using lauroyl peroxide (LPO) as initiator were prepared, and their morphological and CEC properties were studied. The composition of the polymerization mixture (i.e. ratios of monomers/porogenic solvents, 1,4-butanediol/1-propanol and LMA/crosslinker) was optimized. The morphological and chromatographic properties of LMA columns were evaluated by means of SEM pictures and van Deemter plots of PAHs, respectively. The polymerization mixture selected as optimal provided a fast separation of a mixture of PAHs with excellent efficiencies (minimum plate heights of 8.9-11.1 μm). Satisfactory column-to-column (RS D < 4.5%) a…

A hybrid nano-MOF/polymer material for trace analysis of fluoroquinolones in complex matrices at microscale by on-line solid-phase extraction capillary electrophoresis

Abstract A hybrid material (nano-metal organic framework@organic polymer, named as nano-MOF@polymer) was applied for the first time as sorbent for on-line solid-phase extraction capillary electrophoresis with ultraviolet detection (SPE-CE-UV). The resulting material was prepared building layer-by-layer a HKUST-1 (Hong Kong University of Science and Technology-1) nano-MOF onto the polymer surface, which allowed controlling the thickness and maximizing the active surface area. The sorbent was widely characterized at micro- and nano-scale to validate the synthesis and to establish the material properties. Then, fritless microcartridges (2 mm) were assembled by packing only a few micrograms of …

Micellar-organic versus aqueous-organic mobile phases for the screening of β-blockers

A comparative study of the performance of reversed-phase liquid chromatography with micellar-organic (MLC) and aqueous-organic (RPLC) mobile phases is reported for the separation of 16 -blockers (acebutolol, alprenolol, atenolol, bisoprolol, carteolol, celiprolol, esmolol, labetalol, metoprolol, nadolol, oxprenolol, pindolol, practolol, propranolol, sotalol, and timolol). MLC with hybrid mobile phases of sodium dodecyl sulfate (SDS) and propanol is revealed as a very competitive technique for the screening of these drugs. Using a conventional Spherisorb C18 column, the theoretical plates ( N) and asymmetry factors (B/A) for the optimal mobile phase compositions were in the ranges N = 2200–4…

3D printed fluidic platform with in-situ covalently immobilized polymer monolithic column for automatic solid-phase extraction

Abstract In this work, 3D stereolithographic printing is proposed for the first time for the fabrication of fluidic devices aimed at in-situ covalent immobilization of polymer monolithic columns. Integration in advanced flow injection systems capitalized upon programmable flow was realized for fully automatic solid-phase extraction (SPE) and clean-up procedures as a ‘front-end’ to on-line liquid chromatography. The as-fabricated 3D-printed extraction column devices were designed to tolerate the pressure drop of forward-flow fluidic systems when handling large sample volumes as demonstrated by the determination of anti-microbial agents, plastic additives and monomers as models of emerging co…

On the determination of underivatised fatty alcohol ethoxylates by electrospray ionisation–mass spectrometry

Abstract The oligomers of fatty alcohol ethoxylates (FAEs) exhibit large sensitivity differences in mass spectrometry with electrospray ionisation (ESI–MS) and atmospheric pressure chemical ionization (APCI). Standards of the oligomers from m = 1 to 7 ethylene oxide units (EOs) and linear alkyl chains from n = 10 to 18 carbon atoms were infused to examine the relative sensitivities or response factors in several media. The response factors of the [M + H]+ and [M + Na]+ peaks in 9:1 acetonitrile/water and methanol/water media containing acid buffers increased following irregular patterns when n and m increased. In methanol/water the response factors depended on the parity of m, being larger …

Preparation of organic monolithic columns in polytetrafluoroethylene tubes for reversed-phase liquid chromatography

[EN] In this work, a method for the preparation and anchoring of polymeric monoliths in a polytetrafluoroethylene (PTFE) tubing as a column housing for microbore HPLC is described. In order to assure a covalent attachment of the monolith to the inner wall of the PTFE tube, a two-step procedure was developed. Two surface etching reagents, a commercial sodium naphthalene solution (Fluoroetch®), or mixtures of H2O2 and H2SO4, were tried and compared. Then, the obtained hydroxyl groups on the PTFE surface were modified by methacryloylation. Attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy and scanning electron microscopy (SEM) confirmed the successful modification…

Rapid determination of sterols in vegetable oils by CEC using methacrylate ester-based monolithic columns

A method for the determination of sterols in vegetable oils by CEC with UV-Vis detection, using methacrylate ester-based monolithic columns, has been developed. To prepare the columns, polymerization mixtures containing monomers of different hydrophobicities were tried. The influence of composition of polymerization mixture was optimized in terms of porogenic solvent, monomers/porogens and monomer/crosslinker ratios. The composition of the mobile phase was also studied. The optimum monolith was obtained with lauryl methacrylate monomer at 60:40% (wt:wt) lauryl methacrylate/ethylene dimethacrylate ratio and 60 wt% porogens with 20 wt% of 1,4-butanediol (12 wt% 1,4-butanediol in the polymeriz…

According to the CPLL proteome sheriffs, not all aperitifs are created equal!

Combinatorial peptide ligand libraries (CPLLs) have been adopted for investigating the proteome of a popular aperitif in Northern Italy, called "Amaro Branzi", stated to be an infusion of a secret herbal mixture, of which some ingredients are declared on the label, namely Angelica officinalis, Gentiana lutea and orange peel, sweetened by a final addition of honey. In order to assess the genuineness of this commercial liqueur, we have prepared extracts of the three vegetable ingredients, assessed their proteomes, and compared them to the one found in the aperitif. The amaro's proteome was identified via prior capture with CPLLs at two different pH values (2.2 and 4.8). Via mass spectrometry …

Chromium(VI) oxide oxidation of non-ethoxylated and ethoxylated alcohols for determination by electrospray ionization mass spectrometry

A new derivatization procedure to increase the sensitivity of electrospray ionization mass spectrometry (ESI-MS) to non-ethoxylated and ethoxylated alcohols was investigated. The analytes were oxidized with chromium(VI) oxide and the resulting carboxylic and ethoxy-carboxylic acids were isolated by extraction with ethyl acetate; the extracts were alkalinized and infused into the ESI-MS system working in the negative-ion mode. The yields of the combined oxidation-extraction were ca. 100% for non-ethoxylated fatty alcohols dissolved in acetone and they decreased moderately in samples containing increasing amounts of water (e.g., a 75% yield was obtained with 50% water). Ethoxylated alcohols w…

Study of elution behaviour with gradient voltage in CEC using methacrylate monolithic columns.

A theoretical study on the retention behaviour and chromatographic performance of neutral solutes using a lauryl methacrylate-based monolithic column under voltage gradient mode in CEC was carried out. Through a flexible mathematical function based on a modified Gaussian model, the peak shape of compounds was firstly fitted under constant and gradient voltage. Using the peak shape parameters and retention time, the estimation of global chromatographic performance, efficiency and peak capacity under several voltage conditions was performed. The influence of voltage gradient on the separation efficiency is discussed and simple equations are presented to calculate retention and peak widths und…

Characterization of the alcoholic fraction of vegetable oils by derivatization with diphenic anhydride followed by high-performance liquid chromatography with spectrophotometric and mass spectrometric detection

Abstract Aliphatic and triterpene alcohols present in vegetable oils have been identified and determined by HPLC using UV–vis and MS detection after previous derivatization with diphenic anhydride. The alcoholic fraction was obtained by saponification, extraction and TLC (according to the European Union official procedure). Derivatization was performed in tetrahydrofuran in the presence of suspended grinded urea, which increases the reaction rate and yield. Derivatized extracts were chromatographed on a C8 column using gradient elution with acetonitrile/water mixtures containing 0.1% acetic acid, with UV–vis followed by negative-ion mode MS detection. Using linear discriminant analysis of t…

On the use of mixing reactors in flow injection analysis with thermal lens calorimetric detection

Abstract In flow injection analysis with thermal lens calorimetric detection, mixing of reagent streams of different composition before the detection cell produces a large background noise. The noise is due to the heterogeneity of the mixture which produces refractive changes in the probe beam direction. Procedures to measure the homogeneity of the mixture of solutions are derived, and used to evaluate and improve the mixing efficiencies of packed bead reactors. Using well-designed reactors, the mixing noise can be reduced below the noise levels due to other sources.

Preparation of monolithic polymer-magnetite nanoparticle composites into poly(ethylene-co-tetrafluoroethylene) tubes for uses in micro-bore HPLC separation and extraction of phosphorylated compounds

[EN] This paper describes the fabrication of a novel microbore monolithic column modified with magnetite nanoparticles (MNPs) prepared in a poly(ethylene-co-tetrafluoroethylene) (EFTE) tubing, and its application as stationary phase for the chromatographic separation of phosphorylated compounds. In order to obtain the composite column, a two-step procedure was performed. The formation of a glycidyl methacrylate-based monolith inside the activated ETFE tube was firstly carried out. Then, two incorporation approaches of MNPs in monoliths were investigated. The generic polymer was modified with 3-aminopropyltrimethoxysilane (APTMS) to be subsequently attached to MNP surfaces. Alternatively, AP…

Phenolic profiles of olive mill wastewaters treated by membrane filtration systems

BACKGROUND Olive mill wastewater is an important by-product obtained after olive oil extraction. In this investigation, the phenolic composition of olive mill wastewater treated with a semi-industrial membrane filtration system, including ultrafiltration and reverse osmosis modules, was studied. In particular, untreated olive mill wastewater was compared with the permeate and the concentrate fractions of each filtration step. RESULTS 3,4-(dihydroxyphenyl) ethanol and p-(hydroxyphenyl) ethanol were the main compounds of all olive mill wastewater analyzed. A total of 32 compounds, detected at 240 or 280 nm by HPLC-DAD, were considered for phenol quantification. Ultrafiltration reduced phenol …

Capillary Electrophoresis of Free Fatty Acids by Indirect Ultraviolet Detection: Application to the Classification of Vegetable Oils According to Their Botanical Origin

A method for the determination of fatty acids in vegetable oils by capillary electrophoresis with indirect UV–vis detection has been developed. The separation of fatty acids was optimized in terms of Brij surfactant nature and concentration and organic modifier (2-propanol) percentage. The optimal background electrolyte consisted of 10 mM p-hydroxybenzoate, 5 mM Tris at pH 8.8, 80 mM Brij 98, 40% acetonitrile, and 10% 2-propanol. Under these conditions, vegetable oils from five botanical origins (avocado, corn, extra virgin olive, hazelnut, and soybean) were analyzed and the fatty acid contents established. Linear discriminant analysis (LDA) models were constructed using fatty acid peak are…

Cyclodextrins as a Key Piece in Nanostructured Materials: Quantitation and Remediation of Pollutants

Separation and pre-concentration of trace pollutants from their matrix by reversible formation of inclusion complexes has turned into a widely studied field, especially for the benefits provided to different areas. Cyclodextrins are non-toxic oligosaccharides that are well known for their host–guest chemistry, low prices, and negligible environmental impact. Therefore, they have been widely used as chiral selectors and delivery systems in the pharmaceutical and food industry over time. However, their use for extraction purposes is hampered by their high solubility in water. This difficulty is being overcome with a variety of investigations in materials science. The setting-up of novel solid…

Highly Efficient Removal of Neonicotinoid Insecticides by Thioether-Based (Multivariate) Metal–Organic Frameworks

Circumventing the impact of agrochemicals on aquatic environments has become a necessity for health and ecological reasons. Herein, we report the use of a family of five eco-friendly water-stable isoreticular metal-organic frameworks (MOFs), prepared from amino acids, as adsorbents for the removal of neonicotinoid insecticides (thiamethoxam, clothianidin, imidacloprid, acetamiprid, and thiacloprid) from water. Among them, the three MOFs containing thioether-based residues show remarkable removal efficiency. In particular, the novel multivariate MOF {SrIICuII6[(S,S)-methox]1.5[(S,S)-Mecysmox]1.50(OH)2(H2O)}·36H2O (5), featuring narrow functional channels decorated with both -CH2SCH3 and -CH2…

Incorporation of metal-organic framework amino-modified MIL-101 into glycidyl methacrylate monoliths for nano LC separation.

Metal-organic frameworks consisting of amino-modified MIL-101(M: Cr, Al, and Fe) crystals have been synthesized and subsequently incorporated to glycidyl methacrylate monoliths to develop novel stationary phases for nano-liquid chromatography. Two incorporation approaches of these materials in monoliths were explored. The metal-organic framework materials were firstly attached to the pore surface through reaction of epoxy groups present in the parent glycidyl methacrylate-based monolith. Alternatively, NH2 -MIL-101(M) were admixed in the polymerization mixture. Using short time UV-initiated polymerization, monolithic beds with homogenously dispersed metal-organic frameworks were obtained. T…

Quality control of fruit juices by using organic acids determined by capillary zone electrophoresis with poly(vinyl alcohol)-coated bubble cell capillaries

An enhanced method for the determination of organic acids in several fruit juices by capillary zone electrophoresis (CZE) with direct UV-Vis detection has been developed in this work. First, a study with simulated real juice samples was done to find the best separation conditions. Next, several commercial fruit juices were analyzed, and the organic acid contents were quantified in less than 12 min using a poly(vinyl alcohol)-coated fused-silica 'bubble cell' capillary. The present method is reliable, fast and provides detection limits comprised between 0.1 and 2.5 μg mL(-1). Moreover, different chemometric techniques, based on CZE data, were examined. Linear discriminant analysis allowed th…

Smart Materials for Solid-Phase Extraction Applications

Optical recognition and removal of Hg(II) using a new self-chemosensor based on a modified amino-functionalized Al-MOF

Abstract We developed a simple self-chemical optical sensor for the monitoring and removal of ultra-trace levels of Hg(II) from aqueous media. The development of this sensor was based on the covalent attachment of amino-functionalized aluminum-based MOF particles with ninhydrin. The new sensor is densely coated with a chelating ligand to permit an ultra-fast, selective, pH-dependent visualization for removal of Hg(II) with detection limit (LOD∼0.494 μg L−1). Monitoring was accomplished via both a colorimetric signal visible to the naked eye as well as UV–vis absorption spectroscopy. Digital image-based colorimetric analysis has also used as a semi-quantitative analysis for determination the…

Improvement of the electrophoretic protein profiles of Leguminosae gums extracts using gamanase and application to the evaluation of carob–guar mixtures

Abstract A quantitative assay for guar gum in carob gum, based on the extraction of proteins in acetonitrile–water (7:3), separation by capillary electrophoresis and multiple linear regression (MLR) using the areas of nine selected peaks as predictors, was improved by performing the extraction in the presence of gamanase. In the absence of the enzyme, peak migration times and areas depended on the guar content, which complicated peak identification and evaluation. Manual correction of the migration times by comparison with standard electropherograms obtained with pure carob and carob–guar mixtures was required; however, when the proteins were extracted under sonication at 60 °C for 30 min i…

New In-Depth Analytical Approach of the Porcine Seminal Plasma Proteome Reveals Potential Fertility Biomarkers

A complete characterization of the proteome of seminal plasma (SP) is an essential step to understand how SP influences sperm function and fertility after artificial insemination (AI). The purpose of this study was to identify which among characterized proteins in boar SP were differently expressed among AI boars with significantly different fertility outcomes. A total of 872 SP proteins, 390 of them belonging specifically to Sus Scrofa taxonomy, were identified (Experiment 1) by using a novel proteomic approach that combined size exclusion chromatography and solid-phase extraction as prefractionation steps prior to Nano LC-ESI-MS/MS analysis. The SP proteomes of 26 boars showing significan…

Classification of extra virgin olive oils according to their geographical origin using phenolic compound profiles obtained by capillary electrochromatography

Abstract A simple and reliable method for the evaluation of the phenolic fraction of extra virgin olive oils (EVOO) by capillary electrochromatography (CEC) with UV–Vis detection, using lauryl acrylate (LA) ester-based monolithic columns, has been developed. The percentages of the porogenic solvents in the polymerization mixture, and the mobile phase composition, were optimized. The optimum monolith was obtained with a monomers/porogens ratio of 40:60% (wt/wt) using a LA/1,3-butanediol diacrylate ratio of 70:30% (wt/wt) and a 1,4-butanediol/1-propanol ratio of 25:75% (wt/wt). A satisfactory resolution between the phenolic compounds was achieved in less than 25 min with a 15:85 (v/v) ACN–wat…

Determination of the four major surfactant classes in cleaning products by reversed-phase liquid chromatography using serially connected UV and evaporative light-scattering detection.

Abstract A method for the simultaneous determination of the most frequently used surfactant families –linear alkyl benzenesulphonates (LAS), alkyl ether sulphates (AES), fatty alcohol ethoxylates (FAE) and oleins (soaps, fatty acid salts) – in cleaning products, has been developed. The common reversed phase octyl (C8), pentafluorophenyl and biphenyl columns were not capable of separating the anionic LAS and AES classes; however, since only LAS absorbs in the UV, these two classes were independently quantified using a C8 column and serially connected UV and ELSD detection. The best compromise to resolve the four surfactant classes and the oligomers within the classes was achieved with a C8 c…

Micellar liquid chromatography: suitable technique for screening analysis.

The screening capability of micellar liquid chromatography (MLC) is discussed using the reported chromatographic data of several sets of compounds (amino acids, beta-blockers, diuretics, phenethylamines, phenols, polynuclear aromatic hydrocarbons, steroids and sulfonamides) and new results (sulfonamides and steroids). The chromatographic data are treated with an interpretive optimisation resolution procedure to obtain the best separation conditions. Usually, the pH and the concentration of surfactant (sodium dodecyl sulfate, SDS, or cetyltrimethylammonium bromide) for the optimal mobile phase were 2.5-3 and0.12 M, respectively. The nature and concentration of organic solvent depended on the…

Electrokinetic capillary chromatography in a polar continuous‐phase water‐in‐oil microemulsion constituted by water, sodium dodecyl sulfate, and n ‐pentanol

A water-in-oil (w/o) microemulsion (ME) constituted by 15% Tris buffer, pH 8.4, in water and 85% sodium dodecyl sulfate (SDS)/n-pentanol 1:4 mixture, capable of dissolving up to 30% vegetable oils and lard, was used as background electrolyte in reverse microemulsion electrokinetic capillary chromatography (RMEEKC). Owing to the free SDS ions in the continuous phase and some degree of percolation, the ME showed a high conductivity (0.65 mS. cm(-1) at 25 degrees C) and sustained a very stable capillary current. Previous rinsing of the capillary with a quaternary ammonium salt for electroosmotic flow (EOF) reduction, a series of nonionic and anionic solutes dissolved either in the ME or in fat…

Determination ofl-Ascorbic Acid and Total Ascorbic Acid in Vascular and Nonvascular Plants by Capillary Zone Electrophoresis

A capillary zone electrophoresis (CZE) procedure for the determination of l-ascorbic acid (AA) and total ascorbic acid (TAA, as the sum of AA and dehydroascorbic acid) in vascular plants, lichens, bryophytes, and liverworts is described. The samples were frozen in liquid nitrogen and extracted with 2% metaphosphoric acid. To determine AA, an aliquot was directly injected in a fused-silica capillary. The determination of TAA was performed upon its reduction with dl-homocysteine at pH 7. The background electrolyte contained 60 mM sodium chloride, 60 mM sodium dihydrogen phosphate, 1 x 10(-4)% hexadimetrine bromide, and NaOH up to pH 7. The procedure was rapid and highly reproducible; the limi…

CEC column behaviour of butyl and lauryl methacrylate monoliths prepared in non-aqueous media

Polymeric monolithic stationary phases for capillary electrochromatography were prepared using two bulk monomers, butyl methacrylate (BMA) and lauryl methacrylate (LMA), by in situ polymerization in non-aqueous media. The effect of 1,4-butanediol/1-propanol ratio on porous properties was investigated separately for each monomer, keeping the proportion of monomers to pore-forming solvents fixed at 40:60 wt:wt. Also, mixtures of BMA and LMA at different 1,4-butanediol/1-propanol ratios were studied for tailoring the morphological features of the monolithic columns. The chromatographic performance of the different columns was evaluated by means of van Deemter plots of polycyclic aromatic hydro…

Sample streaks and smears in immobilized pH gradient gels

In immobilized pH gradient (IPG) gel formulations as wide as pH 4-9, encompassing neutrality and containing the pK 7.0 acrylamido buffer as one of the buffering ions, smears are directly proportional to the total amount of the pK 7.0 species. At a total level of 10 mM pK 7.0 in these gel formulations, severe smears occur not only for mildly hydrophobic proteins (e.g., recombinant alcalase and termamylase) but also for the relatively hydrophilic pI marker proteins. Streaks and smears are essentially abolished in recipes devoid of the pK 7.0 compound or in formulations containing a maximum of 3 mM of this component. Although partitioning in water/n-octanol has shown the pK 7.0 acrylamido buff…

Solid-phase extraction based on ground methacrylate monolith modified with gold nanoparticles for isolation of proteins

In this study, a novel polymeric material functionalized with gold nanoparticles (AuNPs) was prepared as solid-phase extraction (SPE) sorbent for isolation of proteins. The sorbent was synthesized from a powdered poly(glycidyl-co-ethylene dimethacrylate) monolith, and modified with ammonia, followed by immobilization of AuNPs on the pore surface of the material. To evaluate the performance of this SPE support, proteins were selected as test solutes, being the extraction conditions and other parameters (loading capacity and regenerative ability of sorbent) established. The results indicated that this sorbent could be employed to selectively capture proteins according to their pI, on the basi…

Improving Fractionation of Human Milk Proteins through Calcium Phosphate Coprecipitation and Their Rapid Characterization by Capillary Electrophoresis

This work describes a simple sample pretreatment method for the fractionation of human milk proteins (into their two main groups, whey and caseins) prior to their analysis. The protein-extraction protocol is based on the addition of calcium phosphate to nonadjusted pH human milk. The combination of calcium ions with phosphate results in an effective coprecipitation of caseins. To assess the suitability of this fractionation protocol, the protein extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), LC-MS/MS, and capillary electrophoresis (CE) analysis. The results evidence a significant decrease in contamination of casein fraction with whey proteins…

Use of gold nanoparticle-coated sorbent materials for the selective preconcentration of sulfonylurea herbicides in water samples and determination by capillary liquid chromatography.

Abstract Two new gold nanoparticle (NP) coated materials (silica supported on gold NP with and without ionic liquid) were synthesized for solid phase extraction of sulfonylurea herbicides (SUHs), such as bensulfuron-methyl (BSM), metsulfuron-methyl (MSM), pyrazosulfuron-methyl (PSM), thifensulfuron-methyl (TFM) and triasulfuron (TS), from water samples, followed by capillary liquid chromatography coupled to diode array detection (CLC–DAD). Several factors influencing the preconcentration efficiency of SUHs and its subsequent determination, such as pH of the sample, eluent and reusability of sorbents, have been investigated. Under the optimum conditions, the developed method allowed the dete…

Evaluation of the quality of olive oil using fatty acid profiles by direct infusion electrospray ionization mass spectrometry

Abstract Electrospray ionization mass spectrometry is used to predict the olive oil quality according to European Union marketing standards. Samples were 1:50 diluted in an alkaline 85:15 (v/v) propanol/methanol mixture and directly infused into the electrospray ionization source of an ion trap mass spectrometer. The establishment of ratios of the peak abundances of the free fatty acids followed by linear discriminant analysis was employed to predict the olive oil quality grade. In addition, using multiple linear regression and partial least-squares regression, the percentages of extra virgin and virgin olive oils in binary mixtures were predicted with 5–11% average prediction errors.

A new proposal for the determination of polychlorinated biphenyls in environmental water by using host-guest adsorption

Abstract Polychlorinated biphenyls (PCBs) are ubiquitous environmental pollutants whose wide industrial use has been banned over the years in most countries due to their persistence and bioaccumulation. In fact, the International Agency for Research on Cancer defined them in 2016 as carcinogenic to humans based on sufficient evidence of an increased risk of cancer, being children and pregnant or lactating women the most vulnerable population subgroups. In this work, a new alternative for the determination of polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB138, PCB153, and PCB180) in water samples has been developed by using a cyclodextrin-containing silica microparticulated material as …

In syringe hybrid monoliths modified with gold nanoparticles for selective extraction of glutathione in biological fluids prior to its determination by HPLC.

Abstract In this work, a simple device for extraction glutathione (GSH) in biological fluids using a hybrid monolithic material within a polypropylene syringe is developed. For this purpose, glycidyl methacrylate-based monolith was firstly prepared within this housing material, and the polymer was modified with different ligands (ammonia, cysteamine and cystamine). The resulting materials (containing amine or thiol groups, respectively) were then functionalized with gold nanoparticles (AuNPs). The hybrid material that gave the largest AuNPs coverage was selected as solid-phase (SPE) sorbent and several variables affecting the extraction recovery of this compound were investigated. Under opt…

Characterization of industrial alkylpolyphosphonates by infusion electrospray ionization-ion trap mass spectrometry with identification of the impurities by tandem capillary zone electrophoresis

Technical grade diethylene-triaminepentakis(methylenephosphonic acid) (I), dihexamethylene-triaminepentakis(methylenephosphonic acid) (II), ethylene-diaminetetrakis(methylenephosphonic acid) (III), hexamethylene-diaminetetrakis(methylenephosphonic acid) (IV), amino-tris(methylenephosphonic acid) (V), hydroxyethyl-aminobis(methylenephosphonic acid) (VI), 1-hydroxyethylidene-1,1-diphosphonic acid (VII), and 2-phosphonobutane-1,2,4-tricarboxylic acid (VIII) were characterized by ion trap mass spectrometry with electrospray ionization (ESI-ITMS). Using the negative ion mode and acid and alkaline media, peak series corresponding to the nominal compounds and to impurities with a lower number of p…

Triacylglycerol Analysis in Human Milk and Other Mammalian Species: Small-Scale Sample Preparation, Characterization, and Statistical Classification Using HPLC-ELSD Profiles.

In this work, a method for the separation of triacylglycerols (TAGs) present in human milk and from other mammalian species by reversed-phase high-performance liquid chromatography using a core–shell particle packed column with UV and evaporative light-scattering detectors is described. Under optimal conditions, a mobile phase containing acetonitrile/n-pentanol at 10 °C gave an excellent resolution among more than 50 TAG peaks. A small-scale method for fat extraction in these milks (particularly of interest for human milk samples) using minimal amounts of sample and reagents was also developed. The proposed extraction protocol and the traditional method were compared, giving similar results…

Isolation of human milk whey proteins by solid phase extraction with a polymeric material modified with gold nanoparticles

Abstract This work describes a method for the isolation of human milk whey proteins by solid-phase extraction (SPE) with a polymeric material modified with gold nanoparticles. Human serum albumin, α-lactalbumin, lactoferrin and lysozyme were selected as target proteins to establish the performance of SPE support. Several experimental variables (pH and ionic strength) that affect the SPE protocol were investigated to achieve the maximum extraction efficiency. Under optimal conditions, the SPE sorbent gave excellent recoveries, and offered a high permeability and reusability (> 20 times). The feasibility of this methodology was successfully demonstrated by isolating the target proteins in a m…

Direct infusion mass spectrometry as a fingerprint of protein-binding media used in works of art

A direct infusion mass spectrometry method for the characterization of proteinaceous glues from binding media used in pictorial works of art prior to conservation or restoration treatment is proposed. Amino acids are released by acid hydrolysis and dissolved in a mixture of acidic water and ethanol. This mixture is directly infused into a mass spectrometer without any derivatization. The mass spectrometer is operated in positive ion electrospray mode (ESI-MS) to yield [M+H]+ ions for the amino acids. Relative amounts of each amino acid are calculated for each protein (beef and porcine gelatines, albumin, casein and egg). The analyzed proteins were satisfactorily distinguished. The method is…

Determination of fatty alcohol ethoxylates with diphenic anhydride derivatization and liquid chromatography with spectrophotometric detection

A method for the determination of fatty alcohol ethoxylates (FAEs) using diphenic anhydride as derivatization reagent and RP-HPLC separation with UV-vis detection is presented and compared to derivatization with maleic, phthalic, and other cyclic anhydrides. With these anhydrides, the reaction rates increased when urea was added to the reaction medium, and the yields did not decrease when the samples contained moderate amounts of water. Gradient elution on a C8 column was performed with water/acetonitrile in the presence of 0.1% acetic acid. The use of diphenic anhydride was advantageous for both the chromatographic separation and the detection. Specifically, sensitivity at 200 and 220 nm w…

Determination of tocopherols in vegetable oils by CEC using methacrylate ester-based monolithic columns

The separation and determination of tocopherols (Ts) in vegetable oils by CEC using methacrylate ester-based monolithic columns has been developed. The effects of pore size of the monolithic columns were studied, and the composition of mobile phase was optimized. The optimal pore size of the monolith was obtained with 12 wt% 1,4-butanediol in the polymerization mixture. Excellent resolution between tocopherols was achieved within 10 min analysis time with a 99:1 v/v MeOH-aqueous buffer containing 5 mM tris(hydroxymethyl)aminomethane at pH 8.0. The LODs were lower than 2.3 microg/mL, and interday and column-to-column reproducibilities at 25 microg/mL were better than 5.6%. Using a 93:7 v/v M…

Classification of olive leaves and pulp extracts by comprehensive two-dimensional liquid chromatography of polyphenolic fingerprints.

Abstract The development of a new comprehensive two-dimensional liquid chromatographic method is described, to obtain the profiles of polyphenolic compounds present in olive (Olea europaea L.) leaves and pulps from different genetic origin. Optimisation of the stationary phase nature, particle size, column length and internal diameter, as well as other separation conditions, was performed. Along the study, three stationary phases (C18, PFP and phenyl) in the first dimension (1D), and five (C18, amide, cyano, phenyl and PFP) in the second dimension (2D) were combined to obtain the maximal number of resolved peaks. The optimised method successfully characterised the presence of 26 and 29 comm…

The emerging role of 3D printing in the fabrication of detection systems

Abstract 3D printing is fast evolving as an additive manufacturing technique that has been adopted in (bio)analytical science because of the ample variety of materials and technologies currently available for highly affordable prototyping. This review focuses on the unique characteristics of 3D printing for manufacturing of optical and electrochemical detection systems, and sampling interfaces for analytical purposes using fused deposition modelling, vat polymerization (stereolithography and digital light processing) and photopolymer inkjet printing. The majority of works surveyed within the time span of mid-2018 to mid-2020 encompassed the fabrication of several components of the detection…

Identification of Leguminosae gums and evaluation of carob-guar mixtures by capillary zone electrophoresis of protein extracts.

A procedure for the extraction and capillary zone electrophoresis (CZE) separation of proteins from carob, guar and tara gums in a background electrolyte (BGE) of pH 9 containing 0.1% polyvinyl alcohol is described. The CZE protein profiles exhibit characteristic peaks for each one of the Leguminosae gums, which can be used to construct models capable of identifying samples of carob, guar and tara gums, and predicting the guar content in binary carob-guar mixtures of different geographical origin and harvested in different years. The classification and prediction models are constructed by using linear discriminant analysis (LDA) and multiple linear regression (MLR), respectively. An excelle…

Study of chemical changes produced in virgin olive oils with different phenolic contents during an accelerated storage treatment.

Chemical changes produced in an extra virgin olive oil sample in the presence (EVOO) and absence (EVOOP) of its phenolic fraction during an accelerated storage treatment at 60 degrees C up to 7 weeks were studied. Modifications in phenol content, as well as changes in several quality parameters (free acidity, peroxide value, UV absorbance, fatty acid composition, oxidative stability index, and tocopherol content) were also evaluated under the same storage conditions and compared to those of the same sample deprived of phenolic compounds. When the phenolic extract of the EVOO was studied, a decrease of the antioxidants first present in the sample and an increase of the oxidized products were…

Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability: II. Properties of N-acryloylaminopropanol

The physico-chemical properties and the electrophoretic behavior of the novel set of monomers reported by (Simo-Alfonso et al., Electrophoresis 1996, 17, 723-731) have been evaluated. Of utmost importance was the combination of high hydrophilicity and extreme hydrolytic stability, most desired properties for, any electrophoretic matrix, especially for protein fractionation. One of these monomers (N-acryloylaminopropanol, AAP) was found indeed to be extremely hydrophilic (with a partition coefficient P of only 0.10, vs. P = 0.13 for N-acryloylaminoethoxyethanol and P = 0.20 for acrylamide) and to possess excellent stability to alkaline hydrolysis. Its hydrolysis constant (0.008 L mol-1 min-1…

Classification of pumpkin seed oils according to their species and genetic variety by attenuated total reflection Fourier-transform infrared spectroscopy.

Attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR), followed by multivariate treatment of the spectral data, was used to classify seed oils of the genus Cucurbita (pumpkins) according to their species as C. maxima, C. pepo, and C. moschata. Also, C. moschata seed oils were classified according to their genetic variety as RG, Inivit C-88, and Inivit C-2000. Up to 23 wavelength regions were selected on the spectra, each region corresponding to a peak or shoulder. The normalized absorbance peak areas within these regions were used as predictors. Using linear discriminant analysis (LDA), an excellent resolution among all categories concerning both Cucurbita species a…

Proteomic fingerprinting of mistletoe (Viscum album L.) via combinatorial peptide ligand libraries and mass spectrometry analysis

Abstract Combinatorial peptide ligand libraries (CPLLs), coupled to mass spectrometry (MS) analysis, have been used to investigate in depth the proteome of Viscum album L. (VA), commonly named European mistletoe, in order to provide a first proteomic fingerprinting. For this purpose, the proteins were captured via CPLLs at two different pH values (acidic and neutral). A total of 648 non-redundant proteins were identified by using two different databases. The two pH values, chosen for bead incubations, have contributed to increment the capture ability: 56% and 31% of CPLLs species were respectively recognized at pH 7.2 and at pH 2.2. Finally the biological function of identified proteins was…

Polymer-based materials modified with magnetite nanoparticles for enrichment of phospholipids

[EN] A polymeric material modified with magnetic nanoparticles (MNPs) has been synthesized and evaluated as sorbent both for solid-phase extraction (SPE) and dispersive magnetic solid-phase extraction (MSPE) of phospholipids (PLs) in human milk samples. The synthesized sorbent was characterized by scanning electron microscopy and its iron content was also determined. Several experimental variables that affect the extraction performance (e.g. loading solvent, breakthrough volume and loading capacity) were investigated and a comparison between conventional SPE and MSPE modalities was done. The proposed method was satisfactorily applied to the analysis of PLs in human milk fat extracts in diff…

Single-pump heart-cutting two-dimensional liquid chromatography applied to the determination of fatty alcohol ethoxylates.

Abstract A setup for heart-cutting bi-dimensional liquid chromatography (LC–LC), constructed with a chromatograph provided with a single pump, an auxiliary 6-port 2-position valve (V 6/2 ) and a column selector valve (V CS ), is described. The possible ways of connecting the two valves for LC–LC, namely with V 6/2 first followed by V CS and vice versa, are compared. The possibility of using the setups for preconcentration followed by the backwards transfer of the preconcentrated solutes to the detector or to a second column is also shown. The V 6/2 -first configuration for LC–LC was applied to the characterization of industrial fatty alcohol ethoxylates (FAEs) using UV–vis detection. For th…

Artichoke and Cynar liqueur: two (not quite) entangled proteomes.

Combinatorial peptide ligand libraries (CPLLs) have been adopted to investigate the proteome of artichoke extracts, of a home-made alcoholic infusion and of the Italian Cynar liqueur. The aim of study was not only to perform the deepest investigation so far of the artichoke proteome but also to assess the genuineness of the commercial aperitif via a three-pronged attack. First, different extraction techniques have been used for the characterization of the artichoke's proteome, secondly a home-made infusion has been analyzed and finally the proteome of the commercial drink was checked. The artichoke proteome has been evaluated via prior capture with CPLLs at four different pH (2.2, 4.0, 7.2 …

Enzyme-assisted extraction of proteins from Citrus fruits and prediction of their cultivar using protein profiles obtained by capillary gel electrophoresis

Abstract The suitability of protein profiles established by capillary gel electrophoresis (CGE) as a tool to discriminate between 11 cultivars of Citrus (orange and tangerine) peel and pulp was evaluated in this work. Before CGE analysis, different extraction buffers (which included enzyme-assisted treatments) were compared. The best results were achieved using 5% ( v/v ) Celluclast ® 1.5 L and 5% ( v/v ) Palatase ® 20,000 L buffers for Citrus peel and pulp protein extracts, respectively. The resulting protein profiles obtained were used to construct linear discriminant analysis (LDA) models able to distinguish Citrus peel and pulp samples according to their cultivar. In both cases, all sam…

Classification of vegetable oils according to their botanical origin using n-alkane profiles established by GC-MS.

n-Alkane profiles established by gas chromatography-mass spectrometry (GC-MS) were used to classify vegetable oils according to their botanical origin. The n-alkanes present in corn, grapeseed, hazelnut, olive, peanut and sunflower oils were isolated by means of alkaline hydrolysis followed by silica gel column chromatography of the unsaponifiable fractions. The n-alkane fraction was constituted mainly of n-alkanes in the range C8-C35, although only those most abundant (15 n-alkanes, from 21 to 35 carbon No.) were used as original variables to construct linear discriminant analysis (LDA) models. Ratios of the peak areas selected by pairs were used as predictors. All the oils were correctly …

Use of protein profiles established by CZE to predict the cultivar of olive leaves and pulps

Intact protein profiles established by CZE have been used to predict the cultivar of olive leaves and pulps. For this purpose, proteins were extracted using a mild enzyme-assisted extraction method, which provided higher protein recoveries and a lower environmental impact than other previously described methods. These extracts were subjected to CZE determination under basic conditions using a BGE composed of 50 mM phosphate, 50 mM tetraborate and 0.1% PVA at pH 9. Nine and 14 common peaks, for leaf and pulp samples, respectively, were identified in the nine cultivars studied in this work. In addition, and using linear discriminant analysis of the CZE data, olive leaf and pulp samples belong…

A new mathematical function for describing electrophoretic peaks.

A new model is proposed for characterizing skewed electrophoretic peaks, which is a combination of leading and trailing edge functions, empirically modified to get a rapid recovery of the baseline. The peak model is a sum of square roots and is called thereby "combined square roots (CSR) model". The flexibility of the model was checked on theoretical and experimental peaks with asymmetries in the range of 0-10 (expressed as the ratio of the distance between the center and the trailing edge, and the center and the leading edge of the chromatographic peak, measured at 10% of peak height). Excellent fits were found in all cases. The new model was compared with other three models that have show…

Boronate affinity sorbents based on thiol-functionalized polysiloxane-polymethacrylate composite materials in syringe format for selective extraction of glycopeptides

Abstract In this work, two novel boronate affinity monolithic materials able to extract glycopeptides within a polypropylene syringe are described and compared. The first material was synthesized from glycidyl methacrylate (GMA)-based monoliths modified with poly-3-mercaptopropyl-methylsiloxane (PMPMS) followed by attachment of 4-vinylphenylboronic acid (VPBA) via thiol-ene click reaction. The second material was prepared by using gold nanoparticle (AuNP)-modified monoliths as substrate followed by subsequent attachment of PMPMS and VPBA. The resulting materials were used as sorbents for solid-phase extraction (SPE) to selectively preconcentrate glycopeptides from horseradish peroxidase (HR…

Recent advances in aptamer-based miniaturized extraction approaches in food analysis

Abstract Sample preparation is a relevant step in food analysis to achieve adequate extraction and preconcentration of target analytes before their introduction in the analytical system. Current trends in sample preparation involve moving towards miniaturized extraction devices combined with advanced affinity-based sorbents. Within affinity-based materials, aptamer-based ones have attracted much attention due to their excellent molecular recognition properties, high stability, and the possibility of immobilization onto the surface of different supporting materials, such as nanoparticles, monolithic stationary phases, etc. This review gives an overview of the literature published regarding a…

Preparation and characterization of octadecyl acrylate monoliths for capillary electrochromatography by photochemical, thermal, and chemical initiation†

Monolithic stationary phases based on octadecyl acrylate for CEC using different initiating systems (UV irradiation, thermal, and chemical initiation) in the presence of lauroyl peroxide as initiator were synthesized. For each initiation mode, the influence of the porogenic solvent composition on both the morphological and electrochromatographic properties of the resulting monoliths was investigated. Under optimal conditions, excellent efficiencies for the photochemically and chemically polymerized monoliths (minimum plate heights of 6.9–10.7 and 6.5–12.6 μm, respectively) were achieved. Thermally initiated columns gave lower efficiency values, permeabilities, and longer analysis times comp…

Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability: I. Synthetic route and product characterization.

The novel acrylamido monomer reported by our group (N-acryloylaminoethoxyethanol, AAEE; Chiari et al., Electrophoresis 1994, 15, 177-186), found to combine high hydrophilicity with extraordinary resistance to alkaline hydrolysis, has come under closer scrutiny due to unexpected and random autopolymerization while stored as a 1/1 v/v water solution at 4 degrees C (possibly due to a greater oxidability of the ether group). We have additionally found a unique degradation pathway of the monomer, called "1-6 H-transfer", by which the C1 (on the double bond site), by constantly ramming against the C6, next to the ether oxygen (O7, which in fact favors the transfer of the hydrogen atom by C1), pro…

Determination of alkylphenol ethoxylates by micellar electrokinetic chromatography with bile salts

Octyl- and nonylphenol ethoxylates (OPEs and NPEs) with different numbers of ethoxy units (average values: n = 10 and N = 40 for OPEs, and n = 10 for NPEs) were separated by micellar electrokinetic chromatography under positive polarity using an 80 mM borate buffer of pH 8.5 containing sodium deoxycholate (SDC) or sodium cholate (SC). When sodium dodecyl sulfate (SDS) was added to the background electrolyte (BGE) in the absence of the bile salt, a single peak at a migration time longer than that of the EOF was obtained. Substituting the SDS by a bile salt, the homologues were resolved. At the same bile salt concentration, resolution between the homologues was higher with SDC than using SC. …

Poly(ethylene glycol) diacrylate based monolithic capillary columns for the analysis of polar small solutes by capillary electrochromatography

Monolithic stationary phases based on poly(ethylene glycol) diacrylates for capillary electrochromatography were developed. Several poly(ethylene glycol) diacrylates (Mn 250, 575, and 700) were used as single monomers and the resulting columns were carefully compared. Methanol and ethyl ether were selected as porogenic solvents, and in all cases ultraviolet radiation was selected as initiation method to prepare polymeric monoliths. The influence of the monomer chain length and ratio monomer/porogen on the morphological and electrochromatographic properties of the resulting monoliths was investigated. Several families of compounds with different polarity (alkyl benzenes, organophosphorous pe…

Rapid Evaluation of Oxidized Fatty Acid Concentration in Virgin Olive Oils Using Metal Oxide Semiconductor Sensors and Multiple Linear Regression

This works aims to set up a rapid and nondestructive method to evaluate the advanced oxidation of virgin olive oils (VOOs). An electronic nose based on an array of six metal oxide semiconductor sensors was used, jointly with multiple linear regression (MLR), to predict the oxidized fatty acid (OFA) concentration in VOO samples characterized by different oxidative status. An MLR model constructed using five predictors was able to predict OFA concentration with an average validation error of 9%.

Rapid characterization of alkylpolyphosphonates by CZE with indirect photometric and mass spectrometric detection.

Methods for the rapid characterization of industrial alkylpolyphosphonates (APPs) by infusion MS and CZE with indirect photometric (IPD) and MS detection are described. Technical-grade APPs, including alkylaminepolyphosphonates with 3-5 phosphonate groups and different hydrocarbon skeletons, hydroxyethyl-amino-bis(methylenephosphonic acid), hydroxyethylidene-diphosphonic acid, and 2-phosphonobutane-1,2,4-tricarboxylic acid, were examined. A 10 mM solution of adenosine triphosphate disodium salt at pH 2.2 was used as BGE. The nominal compounds of the industrial APPs and their impurities were well resolved in less than 15 min. The peaks were identified by using extracted ion electropherograms…

Classification of Extra Virgin Olive Oils Produced at La Comunitat Valenciana According to Their Genetic Variety Using Sterol Profiles Established by High-Performance Liquid Chromatography with Mass Spectrometry Detection

A method to classify extra virgin olive oils (EVOOs) according to their genetic variety using sterol profiles obtained by high-performance liquid chromatography (HPLC) with mass spectrometry (MS) detection has been developed. Sterol extracts were chromatographed on a dC18 Atlantis column (100x3 mm, 3 microm) with a gradient of acetonitrile/water (0.01% acetic acid) at a flow rate of 1.0 mL min(-1) and positive-ion mode MS detection. Using linear discriminant analysis of the HPLC-MS data (extracted ion chromatograms), EVOO samples belonging to six genetic varieties cultivated at La Comunitat Valenciana, Spain (Arbequina, Borriolenca, Canetera, Farga, Picual, and Serrana), were correctly clas…

Characterization and quantitation of mixtures of alkyl ether sulfates and carboxylic acids by capillary electrophoresis with indirect photometric detection

The separation, characterization, and determination of mixtures of alkyl ether sulfates (AES) and fatty acids (C10-C16) in background electrolytes (BGEs) containing acetonitrile (ACN)-water mixtures is addressed. Due to inhibition of the ionization of the carboxylate groups, the migration time and the resolution between the fatty acids decreased when the water content of the BGE was reduced, but efficiency and resolution between the AES oligomers improved. The migration times increased and resolution improved by substituting 5% ACN by an equivalent amount of dioxane. A complete separation of the two surfactant classes, up to the AES oligomers with 8 ethylene oxide units (EOs) with respect t…

Determination of antibiotics in meat samples using analytical methodologies: A review

Antibiotics are widely used to prevent or treat some diseases in human and veterinary medicine and also as animal growth promoters. The presence of these compounds in foods derived from food-producing animals can be a risk for human health. Consequently, regulatory agencies have set maximum residue limits for antibiotics in food samples. Therefore, the development of novel methodologies for its determination in food samples is required. Specifically, the analysis and quantification of these substances in meat tissues is a challenge for the analytical chemistry research community. This is due to the complexity of the matrix and the low detection limits required by the regulatory agencies. In…

Performance of a series of novel N-substituted acrylamides in capillary electrophoresis of DNA fragments

DNA separations by capillary electrophoresis in viscous solutions of novel polymers, made with Ω-hydroxyl, N-substituted acrylamides (notably N-acryloyl amino propanol, AAP and N-acryloyl amino butanol, AAB) are evaluated. Whereas in standard poly(acrylamide), at 6% concentration, the theoretical plate number (N) does not exceed 500 000, in 6% poly(AAP) N reaches 922 000 and in 6% poly(AAB) N values as high as 1 200 000 are obtained. Also, copolymers of AAP and AAB give N values in excess of 1 million plates. The two novel monomers (AAP and AAB) remain extremely stable during alkaline hydrolysis and display very good hydrophilicity, while being devoid of the noxious habit of auto-polymeriza…

Classification of extra virgin olive oils according to their genetic variety using linear discriminant analysis of sterol profiles established by ultra-performance liquid chromatography with mass spectrometry detection

Abstract A method to classify extra virgin olive oils (EVOOs) according to their genetic variety using sterol profiles obtained by ultra-performance liquid chromatography (UPLC) with atmospheric pressure chemical ionization mass spectrometry (APCI-MS) detection has been developed. The optimal separation conditions were obtained using a gradient acetonitrile/water (0.01% acetic acid) at a flow rate of 0.8 mL min − 1 and 10 °C. Linear discriminant analysis (LDA) models were constructed with the 11 UPLC-APCI-MS sterol peaks taken from the selective ion recording mode chromatograms. Ratios of the peak areas selected by pairs were used as predictors. With the sequential application of two LDA mo…

Rapid classification of enzymes in cleaning products by hydrolysis, mass spectrometry and linear discriminant analysis

A method for the rapid classification of proteases, lipases, amylases and cellulases used as enhancers in cleaning products, based on precipitation with acetone, hydrolysis with HCl, dilution of the hydrolysates with ethanol, and direct infusion into the electrospray ion source of an ion-trap mass spectrometer, has been developed. The abundances of the ([M+H]+ ions of the amino acids, from the hydrolysates of both the enzyme industrial concentrates and the detergent bases spiked with them, were used to construct linear discriminant analysis models, capable of distinguishing between the enzyme classes. For this purpose, the variables were normalized as follows: (A) the ion abundance of each …

Application of Organic Monolithic Materials to Enantioseparation in Capillary Separation Techniques.

This review article is primarily focused on the state-of-the-art of enantioseparations on organic monolithic materials. The article gives an overview of the chiral stationary phases and its application in capillary electrochromatography (CEC), and capillary- and nano-liquid chromatography (cLC and nLC). Since thousands of publications have been emerged from 2000’s and citing all these papers would extend the scope of this review; then, recent developments from last 10 years (2006 to 2016) will be mentioned. Mostly, stationary phases based on copolymers obtained from chiral functional monomers and surface modifications of organic monoliths with chiral ligands will be discussed. The effective…

Physical chemical properties and kinetics of redox processes in water/soybean oil microemulsions

Microemulsoes de agua em oleo (ME w/o) constituidas de agua, oleo de soja, SDS (dodecil sulfato de sodio) e diversos alcoois de cadeia curta foram caracterizadas sob o ponto de vista fisico-quimico e de propriedades eletroanaliticas. Foram testados diferentes co-surfactantes e razoes surfactante:cosurfactante, sendo a ME com a composicao mais favoravel utilizada para estudar a cinetica de processos redox. Para tanto, realizaram-se medidas de voltametria ciclica utilizando como eletrodo de trabalho um ultramicroeletrodo de disco de Pt e, como eletrodo de referencia e auxiliar, foram usados Ag/AgCl e Pt, respectivamente, e ferroceno como uma sonda. Verificou-se que a estabilidade termodinâmic…

Preparation and evaluation of lauryl methacrylate monoliths with embedded silver nanoparticles for capillary electrochromatography

In this article, capillary columns constituted by lauryl methacrylate monoliths with embedded silver nanoparticles (AgNPs) were developed and tested. Two incorporation approaches of AgNPs in monoliths were explored. The AgNPs were either photogenerated in situ during polymerization of the monolith by UV irradiation, or incorporated to the polymerization mixture (ex situ). The influence of the AgNP concentration on the morphological and chromatographic properties of the polymer matrix was investigated, and both the in situ and ex situ approaches were comparatively discussed. The morphology of the monoliths was characterized by electron microscopic techniques, and their electrochromatographic…

Poly(ethylene glycol) diacrylate-based solid-phase extraction for determination of sulfonamides in meat samples

Abstract In this work, a sorbent based on poly(ethylene glycol) diacrylate (PEGDA) polymer has been synthesized for solid-phase extraction (SPE) of sulfonamides (sulfaguanidine, sulfadiazine, sulfathiazole, sulfapyridine, sulfamethazine, sulfamonomethoxine, sulfamethoxazole, sulfisoxazole) from different meat matrices, which were subsequently determined by HPLC-DAD. Several extraction parameters such as loading and elution solvents as well as other variables (loading capacity, breakthrough volume and reusability) influencing on the analytical performance of the sorbent were optimized. Under the optimal conditions, the developed method was successfully applied to determine the eight sulfonam…

Classification of Pecorino cheeses produced in Italy according to their ripening time and manufacturing technique using Fourier transform infrared spectroscopy

Fourier-transform infrared spectroscopy, followed by linear discriminant analysis of the spectral data, was used to classify Italian Pecorino cheeses according to their ripening time and manufacturing technique. The Fourier transform infrared spectra of the cheeses were divided into 18 regions and the normalized absorbance peak areas within these regions were used as predictors. Linear discriminant analysis models were constructed to classify Pecorino cheeses according to different ripening stages (hard and semi-hard) or according to their manufacturing technique (fossa and nonfossa cheeses). An excellent resolution was achieved according to both ripening time and manufacturing technique. A…

Photografted fluoropolymers as novel chromatographic supports for polymeric monolithic stationary phases

[EN] In this study, porous polymer monoliths were in situ synthesized in fluoropolymers tubing to prepare microbore HPLC columns. To ensure the formation of robust homogeneous polymer monoliths in these housing supports, the inner surface of fluoropolymer tubing was modified in a two-step photografting process. Raman spectroscopy and scanning electron microscopy (SEM) confirmed the successful modification of the inner poly(ethylene-co-tetrafluoroethylene) (ETFE) wall and the subsequent attachment of a monolith onto the wall. Poly(glycidyl methacrylate-co-divinylbenzene), poly(butyl methacrylate-co-ethyleneglycol dimethacrylate) and poly(styrene-co-divinylbenzene) monoliths were in situ synt…

Design, characterization and comparison of materials based on β and γ cyclodextrin covalently connected to microporous silica for environmental analysis

Abstract Determination of organic pollutants in environmental samples presents great difficulties due to the lack of sensitivity and selectivity in many of the existing analytical methods. In this work, the efficiency of materials based on silica structures containing bounded γ-cyclodextrin has been evaluated to determinate phenolic compounds and polycyclic aromatic hydrocarbons in air and water samples, respectively, in comparison with materials made of β-cyclodextrin. According to the results obtained for the material characterization, the new γ-cyclodextrin solid phase does not apparently present any porosity when used in air samples, but it has been shown to work efficiently for the pre…

Chemical Composition, Antioxidant Properties and Antimicrobial Activity of the Essential Oil of Murraya Paniculata Leaves from the Mountains of Central Cuba

The essential oil of Murraya paniculata L leaves from the mountains of the Central Region of Cuba, obtained by hydrodistillation, was analyzed by gas chromatography-mass spectrometry. Eighteen compounds, accounting for 95.1% of the oil were identified. The major component was β-caryophyllene (ca. 30%). The antioxidant activity of essential oil was evaluated against Cucurbita seed oil by peroxide, thiobarbituric acid and p-anisidine methods. The essential oil showed stronger antioxidant activity than that of butylated hydroxyanisole and butylated hydroxytoluene, but lower than that of propyl gallate. Moreover, this antioxidant activity was supported by the complementary antioxidant assay in…

Determination of fatty alcohol ethoxylates and alkylether sulfates by anionic exchange separation, derivatization with a cyclic anhydride and liquid chromatography

A method for the separation, characterization and determination of fatty alcohol ethoxylates (FAE) and alkylether sulfates (AES) in industrial and environmental samples is described. Separation of the two surfactant classes was achieved in a 50:50 methanol-water medium by retaining AES on a strong anionic exchanger (SAX) whereas most FAE were eluted. After washing the SAX cartridges to remove cations, the residual hydrophobic FAE were eluted by increasing methanol to 80%. Finally, AES were eluted using 80:20 and 95:5 methanol-concentrated aqueous HCl mixtures. Methanol and water were removed from the FAE and AES fractions, and the residues were dissolved in 1,4-dioxane. In this medium, este…

Fast Separation and Determination of Sterols in Vegetable Oils by Ultraperformance Liquid Chromatography with Atmospheric Pressure Chemical Ionization Mass Spectrometry Detection

A method for the determination of sterols in vegetable oils by ultraperformance liquid chromatography (UPLC) with atmospheric pressure chemical ionization mass spectrometry detection has been developed. The separation of sterols was optimized in terms of mobile phase composition, column temperature and flow rate. The optimal conditions were achieved using an Acquity UPLC BEH C18 column (50 x 2.1 mm, 1.7 microm) with a mobile phase consistent of acetonitrile/water (0.01% acetic acid) using a linear gradient, at a flow rate of 0.8 mL min(-1) and column temperature of 10 degrees C, giving a total analysis time below 5 min. The determination was performed in selective ion recording mode. The li…

Prediction of wheat dough W and P/L inflation test parameters by capillary zone electrophoresis of a protein extract followed by multivariate regression.

A procedure for the evaluation of the wheat flour hardness, based on capillary electrophoresis of a protein extract in an isoelectric acidic buffer, was developed. The 13 flour samples were extracted twice, and two injections of each extract were made. Separations were performed in a background electrolyte (BGE) containing 40 mM aspartic acid, 6 M urea, and 0.5% hydroxyethylcellulose at 60 degrees C. Using the normalized and corrected areas of 79 peaks and peak groups, a partial least squares regression (PLS1) model was able to predict the flour strength or dough deformation work (W) and the dough tenacity/extensibility ratio (P/L) (Alveograph parameters) with an average relative standard d…

Current trends in affinity-based monoliths in microextraction approaches: A review

Abstract This article reviews the research contributions along the past five years concerning to monolithic materials for the development of affinity-based sorbents in the field of microextraction techniques. The first part of this paper includes an introduction regarding monolithic affinity media and information of different binding agents (such as immunoglobulin-binding proteins, enzymes, lectins, antibodies, aptamers, dyes and immobilized metal ions and nanoparticles, among others) that can act as affinity ligands. Then, the preparation of monoliths and ligand immobilization strategies as well as the different available formats (syringes, pipette tips, spin columns, capillaries, disks an…

Peroxodisulfate as a chemical initiator for methacrylate-ester monolithic columns for capillary electrochromatography

Organic monolithic stationary phases for CEC were synthesized in situ in fused-silica capillaries. Polymerization mixtures were composed of butyl methacrylate, ethylene dimethacrylate, and [2-(methacryloyloxy)ethyl]trimethyl ammonium chloride in the presence of a porogenic solvent, using ammonium peroxodisulfate as chemical initiator, and N,N,N',N'-tetramethylethylenediamine to activate the reaction. The influence of the amount of initiator, temperature, and composition of porogenic solvent on the physical and chromatographic properties of monolithic stationary phases has been investigated. A minimum plate height of 14.5 microm was obtained at 18 wt% of 1,4-butanediol in the polymerization …

Rapid evaluation of oxidised fatty acid concentration in virgin olive oil using Fourier-transform infrared spectroscopy and multiple linear regression

Abstract Fourier-transform infrared spectroscopy (FTIR), followed by multivariate treatment of spectral data, was used to evaluate the oxidised fatty acid (OFA) concentration in virgin olive oil samples characterised by different oxidative status. The entire FTIR spectra (4000–700 cm−1) of oils were divided in 25 wavelength regions. The normalised absorbances of the peak areas within these regions were used as predictors. In order to predict the OFA concentration, multiple linear regression (MLR) models were performed. After a cube root transformation of data, an MLR model constructed using eight predictors was able to predict OFA concentration with an average error of 17%. The main wavelen…

Determination of aerobic-anaerobic metabolism-related compounds in aChaoborus flavicans population by infusion ion trap mass spectrometry of extracts of individual larvae

In a daily migration, the aquatic larvae of Chaoborus flavicans (a phantom midge) alternate oxygen-saturated and anoxic lake strata. To investigate this cycle, larvae were collected at a natural environment, and acetate, propionate, pyruvate, lactate, glycerol, phosphate, maleate, succinate, glucose and citrate were determined. Each larva was homogenized with 200 microL water and deproteinized with a spin-filter; 50 microL aliquots were mixed with 50 microL of a buffer containing 80 mM propylamine, 20 mM HCl and 0.06 mM 2,4-dihydroxybenzoic acid (internal standard) in methanol. The extracts were infused in an electrospray ionization ion-trap mass spectrometer. The limits of detection for th…

Use of triacylglycerol profiles established by high performance liquid chromatography with ultraviolet–visible detection to predict the botanical origin of vegetable oils

A method for the determination of triacylglycerols (TAGs) in vegetable oils from different botanical origins by HPLC with UV–vis detection has been developed. Using a core-shell particle packed column (C18, 2.6 micron), TAG separation was optimized in terms of mobile phase composition and column temperature. Using isocratic elution with acetonitrile/n-pentanol at 10 °C, excellent efficiency with good resolution between most of the TAG peak pairs, within a total analysis time of 15 min, was achieved. Using mass spectrometry detection, a total of 15 peaks, which were common to oils of six different botanical origins (corn, extra virgin olive, grapeseed, hazelnut, peanut and soybean) were iden…

Extraction of β-blockers from urine with a polymeric monolith modified with 1-allyl-3-methylimidazolium chloride in spin column format

Abstract A glycidyl methacrylate-based monolith was modified with imidazolium-based ionic liquid (IL) to be used as stationary phase for solid-phase extraction (SPE). The host monolithic support was prepared by in-situ UV polymerization in spin column format. Two approaches were developed to incorporate the IL into the polymeric monolithic matrix: generation of IL onto the surface monolith, and copolymerization by addition of the IL to the polymerization mixture, which gave the best results. The resulting sorbent materials were morphologically characterized and used for the isolation of five β-blockers from human urine samples. All SPE steps were accomplished by centrifugation, which reduce…

Separation of homologues and isomers of linear alkylbenzenesulfonates by capillary electrophoresis with sodium dodecyl sulfate, carboxylic acids and bile salts.

The ability of several anionic compounds, including carboxylic and dicarboxylic acids, sodium dodecyl sulfate (SDS), and sodium deoxycholate (SDC) and other bile salts, to separate the C(10)-C(13) homologues and the corresponding 20 positional isomers of linear alkylbenzenesulfonates (LAS) by capillary electrophoresis was studied. Up to 19 peaks and a shoulder were observed with a background electrolyte (BGE) containing 10 mM phosphate (pH 6.8), 30% acetonitrile and 40 mM SDS, and 18 peaks were obtained with a BGE containing 10 mM borate (pH 9), 40% ethanol and 40 mM palmitic acid (PA). Resolution increased with the alkyl chain length of the carboxylic acid. Dicarboxylic acids with a short …

Determination of cationic surfactants by capillary zone electrophoresis and micellar electrokinetic chromatography with deoxycholate micelles in the presence of large organic solvent concentrations.

Mixtures of the cationic surfactants benzalkonium chloride (BKC) and cetylpyridinium chloride (CPC) were quickly resolved and reproducibly and reliably determined by using background electrolytes (BGEs) containing 80 mM borate, pH 8.5, bile salts and large concentrations of an organic solvent. When the bile salt is present, the separation mechanism changes from capillary zone electrophoresis (CZE) to a mixed micellar electrokinetic chromatography (MEKC)-CZE, with predominant MEKC interactions, which lead to an excellent resolution of all the solutes, including the C12-C18 homologues of BKC and CPC. A BGE containing 50 mM sodium deoxycholate and 30% ethanol for an extreme resolution, or 20% …

Resolution of overlapped non-absorbing and absorbing solutes using either an absorption null-balance detection window or multivariate deconvolution applied to capillary electrophoresis of anionic surfactants.

Abstract Non-absorbing alkyl ether sulfates (AES) can be separated using anthraquinone-2-carboxylic acid (AQCA) as a probe; however, absorbing alkyl benzene sulfonates (ABS), if present, interfere indirect detection of most AES oligomers. Overcoming of this interference, as well as the simultaneous characterisation and evaluation of AES, fatty acids and ABS, was accomplished by using a diode-array detector and the procedures here discussed. First, it was shown that ABS can be made undetectable by using a 9 nm wide and 227 nm centred charge-absorptivity null-balance detection window (NBDW), where its contribution to the absorbance cancels the dilution effects that its presence induces on the…

Derivatization of hydroxyl functional groups for liquid chromatography and capillary electroseparation

The derivatization reactions commonly used to enhance the analytical signal in the HPLC and CE determination of compounds with hydroxyl functional groups are revised. Focus is placed on the determination of compounds having aliphatic alcohols and phenols while lacking other reactive functional groups. The derivatization with acyl chlorides, organic anhydrides, isocyanates and a variety of other approaches, including oxidation of primary and secondary alcohols, sulfonation, esterification with carboxylic acids, and the use of azides, sulfonyl chlorides and other reagents having miscellaneous leaving groups, is covered. Reactions mainly addressed to introduce a chromophore or a fluorophore in…

Acrylate ester-based monolithic columns for capillary electrochromatography separation of triacylglycerols in vegetable oils

Abstract A simple and reliable method for the evaluation of triacylglycerols (TAGs) in vegetable oils by capillary electrochromatography (CEC) with UV–Vis detection, using octadecyl acrylate (ODA) ester-based monolithic columns, has been developed. The percentages of the porogenic solvents in the polymerization mixture, and the mobile phase composition, were optimized. The optimum monolith was obtained at the following ratios: 40:60% (wt/wt) monomers/porogens, 60:40% (wt/wt) ODA/1,3-butanediol diacrylate and 23:77% (wt/wt) 1,4-butanediol/1-propanol (14 wt% 1,4-butanediol in the polymerization mixture). A satisfactory resolution between TAGs was achieved in less than 12 min with a 65:35 (v/v…

Classification of vegetable oils according to their botanical origin using amino acid profiles established by High Performance Liquid Chromatography with UV–vis detection: A first approach

Abstract A preliminary study using amino acid profiles to classify oils according to their botanical origin has been performed. Amino acid profiles were obtained from hydrolysis of proteins present in vegetable oils, and established by High Performance Liquid Chromatography (HPLC) with UV–vis detection. Proteins present in hazelnut, corn, soybean, olive, avocado, peanut and grapeseed oils were precipitated with acetone, and the residue was hydrolysed in acid medium. The amino acids obtained were derivatized with o -phthaldialdehyde and separated by HPLC. Peaks corresponding to 18 amino acids were observed using a C18 column and a gradient of acetonitrile–water in the presence of a 5 mM citr…

Preparation and characterization of hexyl methacrylate monolithic columns for CEC

The preparation of hexyl methacrylate (HMA) monolithic columns for CEC separations has been investigated with two initiation systems: (i) ammonium peroxodisulphate and TEMED to activate the polymerization reaction, and (ii) by thermal initiation with AIBN. For both initiators, the influence of composition of porogenic solvent on morphological and chromatographic properties of monoliths was investigated. Two porogenic solvent systems, aqueous and non-aqueous media, were also studied for monolithic beds polymerized with AIBN. Under optimal conditions, low minimum plate heights (9.6 mum for peroxodisulphate, 8.4 and 10.0 mum for AIBN in aqueous and non-aqueous porogenic solvents, respectively)…

Recent Advances in Affinity MOF-Based Sorbents with Sample Preparation Purposes

This review summarizes the recent advances concerning metal–organic frameworks (MOFs) modified with several biomolecules (e.g., amino acids, nucleobases, proteins, antibodies, aptamers, etc.) as ligands to prepare affinity-based sorbents for application in the sample preparation field. The preparation and incorporation strategies of these MOF-based affinity materials were described. Additionally, the different types of ligands that can be employed for the synthesis of these biocomposites and their application as sorbents for the selective extraction of molecules and clean-up of complex real samples is reported. The most important features of the developed biocomposites will be discussed thr…

Determination of benzomercaptans in environmental complex samples by combining zeolitic imidazolate framework-8-based solid-phase extraction and high-performance liquid chromatography with UV detection

[EN] In this work, the synthesis of zeolitic imidazolate framework-8 (ZIF-8) crystals and their subsequent application as effective sorbents for extraction and preconcentration of several benzomercaptans from environmental complex samples is described. These materials were prepared by solvothermal approach varying the concentration of n-butylamine modulator to modify the surface of the metal-organic framework. The resulting materials were characterized by scanning and transmission electron microscopy, powder X-ray diffraction and Fourier transform infrared spectroscopy. The ZIF-8 material that gave the best features was selected as extractive phase and the influence of various parameters (s…

Composition, industrial processing and applications of rice bran γ-oryzanol

Abstract Rice bran oil (RBO) (20–25 wt% in rice bran) is a unique rich source of commercially-important bioactive phytochemicals, most of them of interest in nutrition, pharmacy and cosmetics. The unsaponifiable constituents of RBO include mainly tocols (vitamin E, 0.10–0.14%) and γ-oryzanol (esters of trans- ferulic acid with sterols and triterpenic alcohols, 0.9–2.9%). The following topics concerning γ-oryzanol are reviewed: analytical methods for characterisation and determination; influence of genetic and environmental factors on the composition of rice bran; extraction approaches, including supercritical CO 2 and subcritical water; and biomedical and industrial applications, including …

Proteomic fingerprinting of apple fruit, juice, and cider via combinatorial peptide ligand libraries and MS analysis

Combinatorial peptide ligand libraries coupled to MS was applied to extensively map the proteome of apple fruit, and to detect its presence in commercial apple juice and cider to evaluate their authenticity and genuineness. Using the Uniprot_Malus database, 96 proteins were detected in apples, among which 30 proteins were specifically captured via combinatorial peptide ligand libraries. Next, three proteins, previously recognized in fruits, were found in apple juice, which were involved in cellular metabolism of fruit maturation and in allergenic reactions. On the other hand, only one Malus allergen was identified in cider beads eluate, demonstrating that the industrial processes did not pr…

Optimised procedures for the reversed-phase liquid chromatographic analysis of formulations containing tricyclic antidepressants.

The chromatographic behaviour (retention, selectivity, peak shape and resolution) of seven tricyclic antidepressants (TCAs), amitryptiline, clomipramine, doxepin, imipramine, maprotiline, nortryptiline and trimipramine, was examined. Conventional unendcapped Cs and C18 columns and an endcapped XTerra MS C18 column recommended for the analysis of basic compounds were used together with acetonitrile-water and micellar sodium dodecylsulfate (SDS)-pentanol mobile phases. The two best combinations were XTerra C18/acetonitrile, which yielded the largest efficiencies and resolution, and C8/SDS-pentanol, which eliminated the peak tails that were still observed with the XTerra C18 column. Both the s…

Determination of water-soluble vitamins in energy and sport drinks by micellar electrokinetic capillary chromatography

Abstract A method for the determination of water-soluble vitamins in several energy and sport drinks by micellar electrokinetic chromatography (MEKC) has been developed in this work. The separation of vitamins was studied in terms of background electrolyte composition (borate content, pH, surfactant type and content) and in other MEKC parameters. A study of the possible compounds found in the vitamin-enriched drinks that could interfere in vitamin determination was also performed, and a modified procedure with enhanced resolution was developed. The proposed method was successfully applied to the analysis of water-soluble vitamins in a variety of energy and sport drinks and also in fruit nec…

Determination of sulphonamides in human urine by azo dye precolumn derivatization and micellar liquid chromatography

Abstract A high-performance liquid chromatographic method for the determination of sulphonamides in urine is reported. The drugs (sulphadiazine, sulphaguanidine, sulphamethizole, sulphamethoxazole, and sulphathiazole) were diazotized with nitrite and coupled with N-(1-naphthyl)ethylenediamine dihydrochloride in a sodium dodecyl sulphate (SDS) micellar medium. Separation of the sulphonamide azo dyes was performed on a C18 column with a 0.05 M SDS-2.4% pentanol mobile phase, which permitted the direct injection of the urine samples. The limits of detection were in the 0.1–0.3 μg/ml range.

Comparison of thermal&amp;hyphen; and photo&amp;hyphen;polymerization of lauryl methacrylate monolithic columns for CEC

Lauryl methacrylate-based (LMA) monolithic columns for CEC, prepared using either thermal initiation or by UV-irradiation in the presence of AIBN have been compared. Thermal polymerization was carried out at 70 degrees C for 20 h. For UV initiation, the effects of the time exposure to UV light and irradiation energy were investigated. For each initiation process, the influence of composition of porogenic solvent (1,4-butanediol/1-propanol ratio) on the physical and electrochromatographic properties of the resulting monoliths was also evaluated. Photochemically lauryl methacrylate stationary phases initiated showed higher permeabilities and better efficiencies than those prepared by thermal …

Infusion mass spectrometry as a fingerprint to characterize varnishes in oil pictorial artworks.

Mass spectrometry methodology to characterize drying oil used as binding media and varnishes in pictorial artworks, prior to conservation or restoration treatment, is proposed. The analytical treatment requires prior basic hydrolysis of the samples to release the fatty acids : caprylic, pelargonic, capric, sebacic, azelaic, suberic, eicosanoic, lauric, mirystic, palmitic, linolenic, linoleic, oleic and stearic, followed by separation from the matrix by a hexane/water extraction. After removing the solvent, the remaining solid is dissolved in potassium hydroxide, propanol and methanol. The mixture is directly infused into a mass spectrometer without any previous derivatization or separation …

Use of an enzyme-assisted method to improve protein extraction from olive leaves.

The improvement of protein extraction from olive leaves using an enzyme-assisted protocol has been investigated. Using a cellulase enzyme (Celluclast® 1.5L), different parameters that affect the extraction process, such as the influence and amount of organic solvent, enzyme amount, pH and extraction temperature and time, were optimised. The influence of these factors was examined using the standard Bradford assay and the extracted proteins were characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum extraction parameters were: 30% acetonitrile, 5% (v/v) Celluclast® 1.5L at pH 5.0 and 55°C for 15min. Under these conditions, several protein extracts…

High-performance micellar liquid chromatography determination of sulphonamides in pharmaceuticals after azodye precolumn derivatization

Abstract A chromatographic procedure with precolumn derivatization to form the N-(1-naphthyl)ethylenediamine dihydrochloride azodyes is proposed for the analysis of several sulphonamides (sodium sulphacetamide, sulphadiazine, sulphaguanidine, sulphamerazine, sulphamethizole, sulphamethoxazole, sulphanilamide and sulphathiazole) in pharmaceutical preparations (tablets, pills, capsules, suspensions and drops). The separation is performed with a 0.05 M sodium dodecyl sulphate/2.4% pentanol eluent at pH 7. The precolumn derivatization improved the resolution in the chromatograms and increased the selectivity in the determination of mixtures of sulphonamides and in preparations where other drugs…

Micellar liquid Chromatographic determination of diuretics by diazotization and coupling with the Bratton-Marshall reagent

Abstract A Chromatographic procedure that uses micellar mobile phases of sodium dodecyl sulphate (SDS) and propanol at pH 3 is reported for the determination, in urine samples, of thiazide diuretics (althiazide, bendroflumethiazide, chlorothiazide, hydrochlorothiazide, hydroflumethiazide, and trichloromethiazide), and other diuretics having a sulphonamide substituent in the benzene ring. Precolumn derivatization with sodium nitrite and N -(1-naphthyl)ethylenediamine, to form the coloured azo dyes of the hydrolyzed diuretics, decreased the polarity of the compounds and, consequently, their retention in the micellar eluents, and permitted detection in the visible region, eliminating thus the …

Single-pump bi-dimensional LC applied to the characterization of derivatized fatty alcohol ethoxylates.

A column selection valve and a 6-port 2-position injection valve were combined to implement bi-dimensional LC in a system driven by a single pump. The system was applied to the separation of linear fatty alcohol ethoxylates with 12-18 carbon atoms in the hydrocarbon moiety, previous derivatization with diphenic anhydride. Separation on the first separation dimension (C(1)) was carried out with a propyl-diol column and an acetonitrile/aqueous ammonium acetate gradient. Fatty alcohol ethoxylate hydrocarbon series (oligomers with the same number of carbon atoms in the alkyl chain but differing in the number of ethylene oxide units) were resolved without distinguishing the oligomers within the …

Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability: III. DNA separations by capillary electrophoresis in poly(N-acryloylaminopropanol).

Separation of DNA fragments in a novel polymer network, consisting of N-acryloylaminopropanol (AAP) is reported. The performance of this novel monomer, as a sieving liquid polymer in capillary zone electrophoresis, was evaluated. In 50 microns ID capillaries, an 8% solution of poly (AAP) can afford apex-resolution of the 123/124 bp adjacent pair of DNA fragments in marker V, typically unresolved in any poly (acrylamide) formulation. It is proposed that the distal-OH group in the AAP molecule can form transient H-bonds with the DNA double helix. Molecular modeling shows a meandering structure for poly (AAP), lacing the walls of half a cylinder, with kinks protruding at regular intervals, pot…

Direct determination of oleic acid in soybean oil by capacitively coupled contactless conductivity detection capillary electrophoresis in an oil-miscible KOH/1-propanol/methanol medium

Este trabalho teve por objetivo desenvolver um método analítico direto e rápido para a determinação de ácido oleico em óleo de soja por eletroforese capilar com detecção condutométrica sem contato. O eletrólito de corrida empregado foi uma mistura metanol/1-propanol (1:6 v/v) contendo 4 × 10-2 mol L-1 de KOH e 10% (v/v) em etileno glicol. As amostras foram preparadas pela solubilização de 50 g L-1 de óleo de soja e 1,33 × 10-3 de ácido salicílico (padrão interno) no eletrólito de corrida. Os ensaios quantitativos foram realizados adicionando ácido oleico puro às amostras, na faixa entre 0,53 e 2,13 × 10-3 mol L-1. Sob polaridade negativa, os solutos aniônicos deslocaram-se mais rapidamente …

Study of peak shape and efficiency in butyl acrylate-based monolithic columns for capillary electrochromatography.

Abstract The study and modelling peak shape of in capillary electrochromatography (CEC), obtained using butyl acrylate (BA)-based monolithic columns, is described in this article. A modified-Gaussian model, which is a function of conventional experimental parameters: retention time (tR), peak height (H0) and standard deviation (σ0) at the peak maximum, and left (A) and right (B) halfwidths, was used to describe the peaks of thiourea and several polyaromatic hydrocarbons compounds, which were eluted under several applied voltages. A mean relative error below 2% in the prediction of peak shape is obtained. Based on peak parameters, an easy and reliable estimation of global chromatographic per…

Tannin analysis of chestnut bark samples (Castanea sativa Mill.) by HPLC-DAD-MS.

In the present investigation, an HPLC-DAD/ESI-MS method for the complete analysis of tannins and other phenolic compounds of different commercial chestnut bark samples was developed. A total of seven compounds (vescalin, castalin, gallic acid, vescalagin, 1-O-galloyl castalagin, castalagin and ellagic acid) were separated and quantified, being 1-O-galloyl castalagin tentatively identified and found for the first time in chestnut bark samples. Thus, this method provided information regarding the composition and quality of chestnut bark samples, which is required since these samples are commercialised due to their biochemical properties as ingredients of food supplements.