Determination of elemental compositions by gas chromatography - time of flight mass spectrometry using chemical and electron ionization.

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Fatty acid neutral losses observed in tandem mass spectrometry with collision-induced dissociation allows regiochemical assignment of sulfoquinovosyl-diacylglycerols

A full characterization of sulfoquinovosyldiacylglycerols (SQDGs) in the lipid extract of spinach leaves has been achieved using liquid chromatography/electrospray ionization-linear quadrupole ion trap mass spectrometry (MS). Low-energy collision-induced dissociation tandem MS (MS/MS) of the deprotonated species [M - H]- was exploited for a detailed study of sulfolipid fragmentation. Losses of neutral fatty acids from the acyl side chains (i.e. [M - H - RCOOH]-) were found to prevail over ketene losses ([M - H - R'CHCO]-) or carboxylates of long-chain fatty acids ([RCOO]-), as expected for gas-phase acidity of SQDG ions. A new concerted mechanism for RCOOH elimination, based on a charge-remote fragmentation, is proposed. The preferential loss of a fatty acids molecule from the sn-1 position (i.e. [M - H - R1COOH]-) of the glycerol backbone, most likely due to kinetic control of the gas-phase fragmentation process, was exploited for the regiochemical assignment of the investigated sulfolipids. As a result, 24 SQDGs were detected and identified in the lipid extract of spinach leaves, their number and variety being unprecedented in the field of plant sulfolipids. Moreover, the prevailing presence of a palmitic acyl chain (16:0) on the glycerol sn-2 position of spinach SQDGs suggests a prokaryotic or chloroplastic path as the main route for their biosynthesis

Hydrophilic interaction and reversed phase mixed-mode liquid chromatography coupled to high resolution tandem mass spectrometry for polar lipids analysis

A hydrophilic interaction liquid chromatography (HILIC) fused-core column (150 × 2.1 mm ID, 2.7 μm particle size) and a short reversed-phase liquid chromatography (RPLC) column (20 mm × 2.1 mm ID, 1.9 μm) were serially coupled to perform mixed-mode chromatography (MMC) on complex mixtures of phospholipids (PL). Mobile phase composition and gradient elution program were, preliminarily, optimized using a mixture of phosphatidylcholines (PC), phosphatidylethanolamines (PE), their corresponding lyso-forms (LPC and LPE), and sphingomyelins (SM). Thus a mixture of PC extracted from soybean was characterized by MMC coupled to electrospray ionization (ESI) high-resolution Fourier-transform mass spectrometry (FTMS) using an orbital trap analyzer. Several previously undiscovered PC, including positional isomers (i.e. 16:0/19:1 and 19:1/16:0) of PC 35:1 and skeletal isomers (i.e. 18:1/18:2 and 18:0/18:3) of PC 36:3 were identified. Therefore, high-resolution MS/MS spectra unveiled the occurrence of isomers for several overall side chain compositions. The proposed MMC-ESI-FTMS/MS approach revealed an unprecedented capability in disclosing complexity of an actual lipid extract, thus representing a very promising approach to lipidomics

Determination of Maltitol, Isomaltitol, and Lactitol by High-pH Anion-Exchange Chromatography with Pulsed Amperometric Detection

Disaccharide alditols (DAs) such as maltitol, isomaltitol, and lactitol are increasingly being employed in food industry by virtue of their low hydroscopicity, high stability, and good bulking properties. Still, these compounds are reduced-calorie sweeteners, so they are successfully employed in many dietetic foods, like candies, chocolates, baked products, ice creams, and beverages. Here we describe the determination of maltitol, isomaltitol, and lactitol, along with other common carbohydrates, in some foodstuffs such as toffees, biscuits, creams, sponge cakes, chocolates, roasted malt, and chicory leaves. Separations were accomplished by high-pH anion-exchange chromatography (HPAEC) with pulsed amperometric detection using 40 mM NaOH + 1 mM Ba(CH 3 COO) 2 as the mobile phase. The optimal detection potential (E DET ) +0.10 V) was established in voltammetric experiments carried out in batch and flowing stream solutions. Under optimized conditions there was no need for both postcolumn addition of strong bases to the eluent and, even more important, column regeneration between runs. A pellicular column with a relatively low ion-exchange capacity was adopted, which allows a rapid separation of sorbitol, isomaltitol, lactitol, maltitol, glucose, fructose, sucrose, and lactose. The presence in the alkaline mobile phase of barium ions improved selectivity and reproducibility besides shorter analysis times as well. Limits of detection were on the order of 10-20 pmol injected. The contents of DAs and other free sugars in some dietetic foods were evaluated by calibration graphs

Mixed-valent ruthenium oxide - ruthenium cyanide inorganic film on glassy carbon electrodes as an amperometric sensor of aliphatic alcohols

A mixed-valent ruthenium oxide-ruthenium cyanide film on glassy carbon (GC/mvRuO-RuCN) electrode exhibits excellent electrocatalytic activity toward oxidation of simple aliphatic alcohols and polyhydric compounds in acidic media. Electrochemical formation of the ruthenium oxide-based chemically modified electrode can be accomplished by potential cycling or potentiostatic control in diluted sulfuric acid solutions. The attractive electrooxidation capabilities of hydroxyl-containing compounds at this modified electrode are highlighted in terms of sensitivity, stability, and catalytic action. Remarkably, the molar response of the catalytic oxidation increases on increasing the chain length of aliphatic alcohols. For example, the molar response ratio between 1-butanol and methanol is 37 in 25 mM sulfuric acid. Chromatographic separations with electrochemical detection using the GC/mvRuO-RuCN modified electrode allo rr very simple quantitation of aliphatic alcohols in real samples with linear calibration plots over about 3 orders of magnitude. The detection limits for ethanol, 1-propanol, 1-butanol, and 1-pentanol are 4, 0.8, 1, and 2 nmol injected (S/N = 3), respectively

Determination Of Mono- And Disaccharides In Milk And Milk Products By High-Performance Anion-Exchange Chromatography With Pulsed Amperometric Detection

A simple and sensitive liquid chromatographic method for the separation and quantification of mono- and disaccharides in raw- and processed-milk is described. Samples of cows’, buffalos’, sheeps’ and goats’ milk were analyzed upon clarification and appropriate dilution for the quantification of lactose, galactose, glucose and N-acetylglucosamine (GlcNAc). The separation was accomplished by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC–PAD), using a gold working electrode and dilute alkaline eluents modified by a millimolar concentration of barium acetate. The eluent composition employed was designed to provide optimum separation with respect to the selected sample, without interference from the matrix components. The analytical method was successfully employed for the determination of mono- and disaccharides naturally occurring in dairy milk, mozzarella cheese and whey samples, with high sensitivity and accuracy

Profiling of N-Acyl-Homoserine Lactones by Liquid Chromatography Coupled with Electrospray Ionization and a Hybrid Quadrupole Linear-Ion-Trap and Fourier-Transform Ion-Cyclotron-Resonance Mass Spectrometry (LC-ESI-LTQ-FTICR-MS)

A method for the comprehensive profiling of the N-acyl-homoserine lactone (AHL) family of bacterial quorum-sensing molecules is presented using liquid chromatography (LC) coupled to a hybrid quadrupole linear ion trap (LTQ) and Fourier-transform ion-cyclotron-resonance mass spectrometer (FTICR). We demonstrate an increase in signal intensity in MS with electrospray ionization (ESI) of the protonated molecules, [M + H]+, by using acetonitrile (ACN) instead of methanol (MeOH) as the organic solvent under the conditions in which the samples were supplied to the probe by direct infusion at constant flow rates. The presence of ACN prevents the formation of methanol adducts such as [M + MeOH + H]+ and [M + MeOH + Na]+, while also lowering the signal intensity of sodiated [M + Na]+ ions. Sensitivity of these signaling molecules in terms of signal-to-noise ratio (S/N) using low-resolution LTQ-MS and high-resolution FTICR-MS were compared under reversed-phase (RP) LC separations with ESI interface. Special emphasis was paid to the choice of the separation column, its elution conditions and detection of the major AHL compounds produced by the Serratia liquefaciens strain ATCC 27592. The most promising results were obtained using a RP C16-amide column eluted with a linear mobile phase gradient ACN/H2O containing 0.1% formic acid. The whole set of AHL homologs in bacterial extracts was detected in the extracted-ion chromatographic (XIC) mode, and the calculations of molecular formulae were performed by including the isotopic pattern. This mode of displaying data, with a very narrow mass-to-charge ratio window (i.e. ±0.0010 as m/z unit) around each selected ion, has allowed the identification of all the eight known homoserine lactones, viz. C4-HSL, 3-oxo-C6-HSL, C6-HSL, 3-oxo-C8-HSL, C8-HSL, C10-HSL, C12- HSL andC14-HSL. In addition, at least four uncommon signalingmediators previously unreported, namely, 3-oxo-C10 : 1-HSL, 3-oxo-C11 : 2-HSL, 3-oxo-C13 : 2-HSL and 3-OH-C16-HSL, were identified and characterized; their roles in cell-to-cell communication has to be elucidated

Improved Determination of Taurine by High-Performance Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection

As taurine is a very important compound involved in a large number of metabolic processes, it is naturally present in the mammal tissues and is often deliberately added in some foods as a fortifying component. A detailed knowledge of taurine metabolic roles in biological systems can be obtained only if a sensitive, reliable and rapid analytical method is available. This article describes the successful application of high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-IPAD) for taurine determination in egg white and yolk samples, as well extracts of human serum and urine. Applications are shown for determination of taurine in soft drinks and pharmaceutical preparations where the taurine content was evaluated by standard additions. These results were achieved without prior derivatization of taurine

Sensitive quantification of iodide by ion-exchange chromatography with electrochemical detection at a modified platinum electrode

A rapid and very sensitive method for the accurate determination of free iodide in real samples is described. The method is based on anion-exchange chromatographic separation coupled with amperometric detection at a modified platinum electrode under constant applied potential (+0.85 V vs. Ag AgCl). An experimental setup with an in-line and very effective method of electrode modification is proposed using an amperometric thin-layer cross-flow detector and a flowing solution 300 mg/L of iodide; the working electrode is polarised to the limiting current for oxidation of iodide to iodine in acidic solutions with the consequent formation of an iodine-based film. The results indicated that the modified electrode exhibits high analytical response for iodide electrooxidation with good stability and long-life. The signal intensity of daily experimental sessions (8 h), during which standards and real samples were repeatedly injected, exhibits a moderate lowering (i.e. <6%). Using a mixture of 25 mM HNO3 and 50 mM NaNO3 as an eluent phase in ion-exchange chromatography, the detection limit of iodide was estimated to be 0.5 ug/L (S/N=3) with an injection volume of 50 uL. This method was applied successfully to quantify the iodide content of milk samples and in wastewaters as well as trace amounts in common vegetables and solutions containing high chloride levels

Structural characterization of major soyasaponins in traditional cultivars of Fagioli di Sarconi beans investigated by high resolution tandem mass spectrometry

Major soyasaponins, i.e., soyasaponins I, V, βg, and αg from traditional Fagioli di Sarconi beans (Phaseolus vulgaris L., ecotype Tabacchino), were analyzed by reversed-phase liquid chromatography–mass spectrometry (MS) using high-resolution Fourier transform ion cyclotron resonance (FTICR) MS on electrospray ionization in positive-ion mode. Fagioli di Sarconi beans are protected by the European Union [Commission Regulation (EC) No 1263/96] with the mark PGI (for “Protected Geographical Indication”), and are cultivated in Basilicata (southern Italy). Protonated adducts of soyasaponins I, V, βg, and αg were observed at m/z 943.5262, 959.5213, 1069.5583, and 1085.5534, respectively. Gas-phase dissociation of soyasaponins by infrared multiphoton dissociation FTICR MS was performed using a CO2 laser source at a wavelength of 10.6 μm. Most of the fragment ions were identified unambiguously by using the high-resolution and accurate mass value provided by the FTICR mass spectrometer. All soyasaponins exhibit a sequential and neutral loss of sugar moieties at relatively short irradiation times (i.e., less than 50 ms). When the pulse length was increased, a more pronounced fragmentation occurred, with several signals in the lower part of the mass spectrum. In the case of soyasaponins βg and αg, the occurrence of the conjugated product ion at m/z 127.0389 ([C6H6O3 + H]+, 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one) was evidenced. Coupling reversed-phase liquid chromatography with high-performance FTICR MS in combination with infrared multiphoton dissociation tandem MS proved to be very promising for the structural characterization of soyasaponins, and is also suitable for the rapid and accurate structural investigation of other saponins