Characterization of pigments and ligands in a wall painting fragment from Liternum archaeological park (Italy)

Spectroscopic and MS techniques were used to characterize the pigments and the composition of polar and nonpolar binders of a stray wall painting fragment from Liternum (Italy) archaeological excavation. X‐ray fluorescence and diffraction analysis of the decorations indicated mainly the presence of calcite, quartz, hematite, cinnabar, and cuprorivaite. Infrared spectroscopy, GC coupled to flame‐ionization detector, and MS analysis of the polar and nonpolar components extracted from paint layers from three different color regions revealed the presence of free amino acids, sugars, and fatty acids. Interestingly, LC‐MS shotgun analysis of the red painting region showed the presence of αS1‐casein of buffalo origin. Compared to our previous results from Pompeii's wall paintings, even though the Liternum painting mixture contained also binders of animal origin, the data strongly suggest that in both cases a tempera painting technique was utilized.     

Static–dynamic sequential superheated liquid extraction of phenols and fatty acids from alperujo

Superheated liquids of different polarity have been used for sequential extraction of fatty acids and phenols from alperujo. Multivariate methodology has been used to optimise the static–dynamic extraction. Forty-two minutes are required to complete extraction (20 mg/kg of fatty acids and up to 2,200 mg/kg of hydroxytyrosol in the raw material used). The efficacy of the extraction has been demonstrated and compared with that of conventional methods (Folch and stirring-based methods for fatty acids and phenols, respectively), which needed 4.5 and 24 h for the extraction of fatty acids and phenols, respectively. The non-polar and polar extracts were injected into GC–MS and HPLC–MS–MS equipment, respectively, for individual separation–quantification of the target compounds. The simplicity of the experimental setup and the low costs of the raw material make the proposed method advisable when extraction of both fractions is required.     

Thermodynamic parameters of the interaction of cryptand[222] with amino acids in water at 298.15 K

The interaction of cryptand[222] with amino acids in water, which is weak for most amino acids and controlled by the solvent effect in the case of non-polar amino acids, was studied at 298.15 K by the calorimetric method. Cryptand[222] undergoes selective complex formation with some polar and aromatic amino acids. The thermodynamic functions and equilibrium constants of complex formation of the macrocyclic ligand withl-histidine,l-threonine, andl-glutamine were calculated. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 12, pp. 2285–2288, December, 1999.     

Characterization of medieval proteinaceous painting media using gas chromatography and gas chromatography — mass spectrometry

Proteinaceous organic materials used as ancient painting media were investigated by capillary gas chromatography (GC) and capillary gas chromatography — mass spectrometry (GC/MS). Medieval wall paintings made by the tempera technique were considered and their binding media were studied by the characterization of their main chemical components. The basic methodology is based on the determination of amino acids in samples of paint layers after hydrolysis and derivatization and on the comparison with reference proteinaceous materials. Multivariate chemometric techniques were used to facilitate the recognition of the protein source from chromatographic data. To characterize the binders further, a method was developed for the determination of fatty acids, present as minor components, by GC/MS. The use of fused-silica capillary columns coated with selected stationary phases allowed the separation of amino acid and fatty acid derivatives in a single analytical run.     

Prediction of retention indexes. III. Silylated derivatives of polar compounds.

Polar compounds containing hydroxyl, amino and carboxyl groups, singly or in combination, can be chromatographed after the polar functional groups are silylated. The silylated derivatives of acids, alcohols, amines, diols, amino alcohols, amino acids are shown to behave chromatographically as hydrocarbons, and their retention indexes can be readily predicted from their base values. The column difference, namely, the difference between the retention indexes of the analyte on polar and non-polar columns is minimal for the silylated derivatives in comparison to that observed for the underivatized analytes. This minimal column difference is attributed to the hydrocarbon-like chromatographic characteristics of the silylated derivatives. The retention indexes of the silyl derivatives appear to correlate with the atom number Z of the analyte.     

Tentative identification of polar and mid‐polar compounds in extracts from wine lees by liquid chromatography–tandem mass spectrometry in high‐resolution mode

Sustainable agriculture has a pending goal in the revalorization of agrofood residues. Wine lees are an abundant residue in the oenological industry. This residue, so far, has been used to obtain tartaric acid or pigments but not for being qualitatively characterized as a source of polar and mid‐polar compounds such as flavonoids, phenols and essential amino acids. Lees extracts from 11 Spanish wineries have been analyzed by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) in high resolution mode. The high‐resolution power of LC–MS/MS has led to the tentative identification of the most representative compounds present in wine lees, comprising primary amino acids, anthocyans, flavanols, flavonols, flavones and non‐flavonoid phenolic compounds, among others. Attending to the profile and content of polar and mid‐polar compounds in wine lees, this study underlines the potential of wine lees as an exploitable source to isolate interesting compounds. Copyright © 2015 John Wiley & Sons, Ltd.     

Influence of solvent polarity on the separation of leucine enantiomers by β-cyclodextrin: a molecular mechanics and dynamics simulation

The interaction between leucine and β-cyclodextrin with different solvents was studied by molecular mechanics and dynamics simulations. In order to analyse the influence of the solvent polarity on the inclusion complex formation and separation process of leucine enantiomers by β-cyclodextrin, the organic modifiers were characterised by the same value of dielectric constant in the electrostatic contribution to the interaction energy, and a different molecular configuration of amino acids (neutral or zwitterion). The complexes formed in polar solvents were more stable than those in non-polar solvents with the same dielectric constant, because the electrostatic contribution is negative for the former and positive for the latter. The optimized structures obtained for leucine enantiomers and β-cyclodextrin in vacuo are non-inclusion complexes. The solvent polarity contributes to increasing the probability of the presence in an inner position for the guest, whereas the results for non-polar configurations were smaller and distributed in larger areas. The regions where the enantiomers spend more time in the simulation correspond to locations with greater chiral discrimination. d-Leu was the first eluted enantiomer in every case, except for a polar solvent with $\epsilon =26$.     

Fractionation of casein hydrolysates using polysulfone ultrafiltration hollow fiber membranes

The objective of this study was to characterize the fractionation profile of casein hydrolysates obtained with polysulfone hollow fiber ultrafiltration membranes. The two-step ultrafiltration process developed by Turgeon and Gauthier [J. Food Sci., 55 (1990) 106] was used: a caseinate solution was submitted to proteolysis with chymotrypsin or trypsin, and the reaction mixture (RM) was subsequently ultrafiltered using a 30 kDa (MWCO) hollow-fiber polysulfone membrane. The total hydrolysate permeating from this first step was further fractionated using a 1 kDa (MWCO) membrane, producing the mixture of polypeptides (retentate) and the amino acid fraction (permeate). The effect of enzyme specificity and of membrane retentivitiy on the total composition (total nitrogen, fat, lactose, minerals) and amino acid profile of the fractions was studied. The overall composition of the fractions was not significantly affected by the nature of the enzyme but the degree of hydrolysis and the molecular weight distribution profile analyses showed a marked effect of the enzyme specificity, with trypsin giving a larger proportion of small peptides (< 200 Da) in the mixture of polypeptides. Amino acid profile analyses provided useful information on the phenomena governing the fractionation of amino acids with a polysulfone membrane: (1) the target amino acids of the enzyme are concentrated in the permeate as a result of their presence in all peptides produced by hydrolysis, (2) polar amino acids are retained by the membrane, (3) non-polar amino acids are not selectively rejected by the membrane. Our results suggest that the charge/hydrophobicity balance of the peptides produced is the predominant factor determining the fractionation of casein hydrolysates.     

Solid-phase extraction element based on epoxy polymer monolith for determination of polar organic compounds in aqueous media

A solid-phase extraction element based on epoxy polymer monolith was fabricated for sorptive enrichment of polar compounds from liquid and gaseous samples. After ultrasonication of the element in an aqueous solution for a given period of time, the thermal desorption (TD) using a pyrolyzer with gas chromatography/mass spectrometry (GC/MS), in which TD temperature was programmed from 50 to 250 °C for the analytes absorbed in the element, was used to evaluate the element for basic extraction performance using the aqueous standard mixtures consisting of compounds having varied polarities such as hexanol, isoamyl acetate, linalool, furfural and decanoic acid, in concentrations ranging from 10 μg/L to 1 mg/L. Excellent linear relationships were observed for all compounds in the standard mixture, except decanoic acid. In the extraction of beverages such as red wine, the extraction element showed stronger adsorption characteristics for polar compounds such as alcohols and acids than a non-polar polydimethylsiloxane-based element. This feature is derived from the main polymer structure along with hydroxyl and amino groups present in the epoxy-based monolith polymer matrix.     

MALDI TOF/TOF tandem mass spectrometry as a new tool for amino acid analysis

This is the first report of an application of collisionally induced fragmentation of amino acids (AA) and their derivatives by MALDI TOF/TOF tandem mass spectrometry (MS). In this work, we collected the data on high-energy fragmentation reactions of a large group of protonated amino acids and their derivatives with the goal of determining which product ions are analyte specific and if yields of these fragment could be used for quantitative analysis. From 34 different amino acids (20 alpha-amino acids, beta-amino acids, homocysteine, GABA, and modified AA Met sulfone and sulfoxide, hydroxyproline, etc.) we observed that high yields of the target specific immonium ions and fragmentation patterns are most similar to EI or FAB CID on sector instruments. The major exceptions were two highly basic amino acids, Arg and Orn. It is noted that neither beta-, gamma-, nor delta-amino acids produce immonium ions. As might be predicted from high-energy CID work on peptides from the sectors and TOF/TOF, the presence of specific indicator ions in MALDI tandem MS allows distinguishing isomeric and isobaric amino acids. These indicator ions, in combination with careful control of data acquisition, ensure quantitative analysis of amino acids. We believe our data provide strong basis for the application of MALDI TOF/TOF MS/MS in qualitative and quantitative analysis of amino and organic acids, including application in clinical medicine.     

Spectrometric analysis,phenolics isolation and cytotoxic activity of <Emphasis Type="Italic">Stipagrostis plumosa</Emphasis> (Family Poaceae)

Grasses (family Poaceae) are economically important plants; they are used as crops and animal foods. Stipagrostis plumosa (L.) Munro ex T. Anderson is a member of this family and subjected to chemical and biological studies. The chromatographic techniques,  LC–ESI–MS and GC/MS were used for identification of polar and non-polar compounds in its extract. Ten compounds, including one new flavone glycoside; tricin 7-O-galactoside, three known flavones, three C-glycosyl flavones and three phenolic acids, were isolated from S. plumosa for the first time except tricin. Their structures were elucidated on the basis of extensive spectroscopic interpretation. In addition to the isolated compounds, eleven compounds were tentatively identified using LC–ESI–MS, five of them were detected for the first time from this species. 29 non polar compounds were identified using GC–MS analysis, representing 83.13% of S. plumosa diethyl ether extract. In addition to the DPPH activity evaluation, the crude extract and the isolated compounds were investigated against five human carcinoma cell lines; A549, HCT-116, HepG2, MCF-7 and PC3 at a concentration of 100 μg/ml. From the isolated compounds tricin and luteolin 6,8-di-C-glucoside could be considered as natural-free radical scavenging agents.     

Experimental study on solvent-less sample preparation methods: Membrane extraction with a sorbent interface, thermal membrane desorption application and purge-and-trap

Different solvent-free sample preparation techniques for the enrichment of volatile and semivolatile organic compounds from aqueous samples for subsequent gas chromatographic separation and detection are compared. The methods under study are purge-and-trap, membrane extraction with a sorbent interface in two different configurations, and thermal membrane desorption application. The study has been performed with polar as well as with non-polar compounds in respect to sampling yield, enrichment, repeatability and analysis cycle rate. All experiments have been performed with a mobile GC–MS system.     

Cyano-modified pre-coated plates — a medium polar modification for high performance thin-layer chromatography

Summary The introduction of a cyano-modified, pre-coated layer substantially widens the selectivity of stationary phases in thin-layer chromatography. This is a moderately polar sorbent based on silica gel 60, which can be used both in adsorption chromatography and in reversed-phase chromatography. This new pre-coated layer is particularly suitable for separation of steroids, alkaloids and derivatized amino acids. The possibility of separating habitforming drugs and preservatives in the presence of ionpair reagents is also discussed.     

Certification of NIST standard reference material 2389a, amino acids in 0.1 mol/L HCl—quantification by ID LC-MS/MS

An isotope-dilution liquid chromatography-tandem mass spectrometry (ID LC-MS/MS) measurement procedure was developed to accurately quantify amino acid concentrations in National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 2389a—amino acids in 0.1 mol/L hydrochloric acid. Seventeen amino acids were quantified using selected reaction monitoring on a triple quadrupole mass spectrometer. LC-MS/MS results were compared to gravimetric measurements from the preparation of SRM 2389a—a reference material developed at NIST and intended for use in intra-laboratory calibrations and quality control. Quantitative mass spectrometry results and gravimetric values were statistically combined into NIST-certified mass fraction values with associated uncertainty estimates. Coefficients of variation (CV) for the repeatability of the LC-MS/MS measurements among amino acids ranged from 0.33% to 2.7% with an average CV of 1.2%. Average relative expanded uncertainty of the certified values including Types A and B uncertainties was 3.5%. Mean accuracy of the LC-MS/MS measurements with gravimetric preparation values agreed to within |1.1|% for all amino acids. NIST SRM 2389a will be available for characterization of routine methods for amino acid analysis and serves as a standard for higher-order measurement traceability. This is the first time an ID LC-MS/MS methodology has been applied for quantifying amino acids in a NIST SRM material.     

High-performance liquid chromatography of nucleic acid constituents: Chromatographic examination of novel stationary phases

Summary Silica-bonded stationary phases were developed for the separation of nucleic acid constituents and their properties investigated with homologous oligoriboadenylic acids in electrostatic interaction chromatography and with alkylbenzenes in reversed-phase chromatography. Analysis of retention data confirmed the stratified molecular structure of the surface which consist of a layer of propyl chains anchoredvia siloxane bridges to the silica surface proper and of polar moieties attached to the hydrocarbonaceous functions. The polar top layer contains weak cationic and/or hydrophobic binding sites, is strongly hydrated in contact with aqueous eluents and bars the access by large biopolymers to the hydrocarbonaceous sublayer. In reversed-phase chromatography of small non polar molecules with hydro-organic eluents, however, this layer is accessible and engenders a retentive behavior typical for weak hydro-carbonaceous bonded phases. As a result the stationary phases, depending on the nature of the sample and the mobile phase, exhibit the properties of "soft" phases for the chromatography of biopolymers under mild elution conditions and those of "hard" phases for the separation of small non-polar molecules under conditions generally employed in reversed-phase chromatography. The retention of nucleic acid constituents on most of the stationary phases investigated subject to a dual mechanism as a result of the interplay of electrostatic and hydrophobic interactions between the eluites and the binding sites on the stationary phase surface. Siliceous stationary phases having surface morphology described above are suitable for the separation of nucleic acid constituents having widely ranging molecular weights up to 3 × 10⁶ Daltons provided the support has appropriate pore dimensions. This is demonstrated by the separation of mixtures arising from digesting t-RNA^pha or polyadenylic acids as well as those of ribosomal RNA’s and different forms of the plasmid pBR322 DNA. Presented at the 15th International Symposium on Chromatography, Nürnberg, October 1984     

Combining chip-ESI with APLI (cESILI) as a multimode source for analysis of complex mixtures with ultrahigh-resolution mass spectrometry

Recently we have established atmospheric-pressure laser ionisation (APLI) as a method for coupling time-of-flight mass spectrometric detectors (TOF MS) with chromatographic systems (HPLC and GC) to allow two-photon ionisation of non-polar aromatic compounds. Here we demonstrate that APLI can be combined with chip-electrospray ionisation (cESI) coupled to Fourier-transform-ion cyclotron resonance mass spectrometry (FT-ICR MS) for ultrahigh-resolution analysis of complex samples. With the laser turned off, the analytes are ionised only by ESI, whereas when the laser is switched on non-polar aromatic substances also are ionised. In combination with the extremely high mass resolution of an FT-ICR MS, simultaneous qualitative analysis of polar and non-polar analytes is possible in both positive and negative modes, as is exemplified with a crude oil sample. Nevertheless, ion suppression was observed (up to ca. 70% for D₁₀-pyrene) and thus sample preparation with chromatographic or electrophoretic pre-separation is necessary for quantitative analysis of targets. In addition, for the first time, the dopant-assisted APLI method in combination with cESI (DA-cESILI) was used for determination of 1-nitrocoronene. Philippe Schmitt-Kopplin, Klaus J. Brockmann and Oliver J. Schmitz contributed equally to the work.     

Influence of additives on the clouding phenomenon of chlorpromazine hydrochloride solutions

Herein we report the effect of various additives (viz. alcohols, cycloalcohols, amino acids, sugars, ureas) on the clouding phenomenon observed in 50mM chlorpromazine hydrochloride (CPZ) drug solutions (prepared in 10mM sodium phosphate buffer). Long chain alcohols (except octanol), cyclohexanol and allylalcohol increased the cloud point (CP) followed by a decrease with the increase in alcohol concentration but short chain alcohols affected the CP insignificantly. Effect of amino acids depended upon their nature: acidic and salts of basic amino acids increased the CP while basic amino acids depressed it; non-polar and uncharged polar amino acids caused small changes in CP. Additives of urea family decreased the CP. All sugars caused a decrease in CP, which is in consonance to their effect on the critical micellar concentration. The overall behavior is explained on the basis of additives affecting the solvent as well as micelle aggregation and/or structure.     

Surface-assisted laser desorption/ionization-mass spectrometry using TiO2-coated steel targets for the analysis of small molecules

Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI–MS) measurements in the low-molecular-mass region, ranging from 0 to 1000 Daltons are very often difficult to perform because of signal interferences originating from matrix ions. In order to overcome this problem, a stainless steel target was coated with a homogeneous titanium dioxide layer. The layer obtained was further investigated for its ability to desorb small molecules, e.g., amino acids, sugars, poly(ethylene glycol) (PEG) 200, or extracts from Cynara scolymus leaves. The stability of the layer was determined by repeated measurements on the same target location, which was monitored by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) before and after surface-assisted laser desorption/ionization (SALDI) analysis. In addition, this titanium dioxide layer was compared with an already published method with titanium dioxide nanopowder as inorganic matrix. As a result of this work, the titanium dioxide layer produced minimal background interference, enabling simple interpretation of the detected mass spectra. Furthermore, the TiO₂ coating provides a target that can be reused many times for SALDI–MS measurements.     

Hydrophilic interaction chromatography using amino and silica columns for the determination of polar pharmaceuticals and impurities

Hydrophilic interaction chromatography (HILIC) is described as a useful alternative to reversed-phase chromatography for applications involving polar compounds. In the HILIC mode, an aqueous-organic mobile phase is used with a polar stationary phase to provide normal-phase retention behavior. Silica and amino columns with aqueous-acetonitrile mobile phases offer potential for use in the HILIC mode. An examination of the retention and separation of several pyrimidines, purines, and amides on silica and amino columns from three manufacturers revealed that mobile phases should contain a buffer or acid for pH control to achieve similar and reproducible results among columns from different sources. Amino columns may also be used in an anion-exchange mode, which provides an advantage for some applications. In some cases, silica can provide different selectivity and better separation than an amino column. Example applications include: low-molecular-mass organic acids and amides as impurities in non-polar drug substances, 5-fluorouracil in 5-fluorocytosine, guanine in acyclovir, and different selectivity for polar basic compounds compared to an ion-pairing system.