Positive Mass Spectrometry Research Articles

Monolignol lithium cation basicity estimates and lithium adduct ion optimized geometries

Mass spectrometric analysis of lignin for developing biomaterials requires advances of characterization techniques. Positive ion mass spectrometry of lignin model compounds using lithium has recently been explored as a viable alternative to current negative mode techniques. To date, little is known about the impact of lithium adduct ion formation on relative response factors of lignin and lignin decomposition products. In this contribution, we report estimates of lithium cation basicity for synthetic monolignols H, G and S using Cooks’ kinetic method on a linear quadrupole ion trap mass spectrometer. Optimized geometries and interaction energies have also been calculated by DFT methods to quantify the electrostatic cation coordination. Based on a combination of experimental and computational evidence, lithium appears to preferentially bind to the phenol and methoxy substituents on the aromatic ring of monolignols. The strength of this interaction increases with the number of methoxy substituents (S > G > H). This work serves as a basis of understanding for future work in developing lithium adducted lignin mass spectrometric analytical methods.

Characterization and sequencing of lithium cationized β-O-4 lignin oligomers using higher-energy collisional dissociation mass spectrometry

The prospect of developing new bio-based products from lignin partially depends on the development of effective analytical techniques to characterize the end products from lignin degradation experiments. To date, the most utilized mass spectrometric technique for characterizing lignin oligomers has been negative ion mass spectrometry. Positive ion mass spectrometry remains a relatively unexplored approach for lignin sequencing. Here we report on the sequencing of lignin oligomers using lithium cationization positive ion electrospray followed by higher-energy collisional dissociation (HCD) tandem mass spectrometry on a high-resolution accurate-mass Orbitrap mass spectrometer. Thirteen synthetic lignin model oligomers containing β-O-4 and 4-O-α linkages were analyzed and sequenced based on their HCD tandem mass spectrometry fragmentation patterns. Proposed sequence-specific fragmentation pathways are provided. The method presented in this study provides a potential application of lithium cationization tandem mass spectrometry for the analysis of lignin degradation products.

Mass spectrometry for characterization of homologous piperidine alkaloids and their activity as acetylcholinesterase inhibitors.

Piperidine alkaloids from Senna spectabilis constitute a rare class of natural products with several biological activities. However, the absence of chromophores makes their structural elucidation by conventional methods a great challenge. In this context, mass spectrometry emerges as a powerful tool for metabolomics studies. The piperidine alkaloids (-)-cassine and (-)-spectaline and the semisynthetic derivatives (-)-3-O-acetylcassine and (-)-3-O-acetylspectaline were investigated by electrospray ionization tandem mass spectrometry (ESI-MS/MS) in the positive mode and electron ionization mass spectrometry (EI-MS). ESI fragmentation studies were performed with a quadrupole time-of-flight instrument; N2 was used as collision gas. The acetylcholinesterase inhibitory activity of the investigated compounds was evaluated by bioautography and microplate screening assays. ESI-MS/MS and EI-MS provided valuable and complementary information about the structure of the piperidine compounds. Collision-induced dissociation experiments (MS/MS) revealed that neutral elimination of water or acetic acid is the major fragmentation pathway, which agrees with the stereochemistry proposed for (-)-cassine and (-)-spectaline and the semisynthetic derivatives (-)-3-O-acetylcassine and (-)-3-O-acetylspectaline. The ESI-MS/MS and EI-MS studies allowed us to propose fragmentation mechanisms for piperidine alkaloids and derivatives. Therefore, mass spectrometry is an important tool for characterizing the structure of these compounds and for supporting further metabolomics studies.

Liquid chromatography–tandem MS/MS method for simultaneous quantification of paracetamol, chlorzoxazone and aceclofenac in human plasma: An application to a clinical pharmacokinetic study

A facile, fast and specific method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous quantitation of paracetamol, chlorzoxazone and aceclofenac in human plasma was developed and validated. Sample preparation was achieved by liquid-liquid extraction. The analysis was performed on a reversed-phase C18 HPLC column (5 μm, 4.6 × 50 mm) using acetonitrile-10 mM ammonium formate pH 3.0 (65:35, v/v) as the mobile phase where atrovastatin was used as an internal standard. A very small injection volume (3 μL) was applied and the run time was 2.0 min. The detection was carried out by electrospray positive and negative ionization mass spectrometry in the multiple-reaction monitoring mode. The developed method was capable of determining the analytes over the concentration ranges of 0.03-30.0, 0.015-15.00 and 0.15-15.00 μg/mL for paracetamol, chlorzoxazone and aceclofenac, respectively. Intraday and interday precisions (as coefficient of variation) were found to be ≤12.3% with an accuracy (as relative error) of ±5.0%. The method was successfully applied to a pharmacokinetic study of the three analytes after being orally administered to six healthy volunteers.

Coupling supercritical fluid chromatography to positive ion atmospheric pressure ionization mass spectrometry: Ionization optimization of halogenated environmental contaminants

Currently used analytical techniques for halogenated aromatic environmental contaminants such as polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs), also known as legacy persistent organic pollutants, are based on gas chromatographic separation of target analytes and detection by mass spectrometry. The coupling of packed column supercritical fluid chromatography (SFC) to atmospheric pressure ionization mass spectrometry (API/MS) could allow for the concurrent analysis of thermally labile and legacy halogenated environmental contaminants if ionization can be sufficiently optimized. The evaluation of positive ion atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) as well as possible charge transfer dopants for the generation of molecular ion isotopomeric clusters of halogenated environmental contaminants with minimal fragmentation has been completed. Using the investigated parameters, positive ion APPI was found to be the more sensitive technique. Of the aromatic and cycloalkane dopants investigated, only fluorobenzene and trifluorotoluene were found to be effective dopants for the halogenated aromatic target analytes (PCDDs, PCDFs, and PCBs). Experiments involving deuterated dopants confirmed that reactive species generated by cycloalkanes were quenched by the SFC eluent rendering them unusable in conjunction with the investigated separation technique. Alternatively, aromatic dopants were found to be less susceptible to quenching by the SFC eluent and fluorobenzene was determined to be the most effective charge transfer dopant for PCDDs, PCDFs, and PCBs. To demonstrate the applicability of the optimized ionization conditions, SFC-API/MS has been used for the concurrent analysis of legacy halogenated aromatic environmental contaminants (PCDDs, PCDFs, and PCBs) and thermally labile analytes (α, β, and γ isomers of hexabromocyclododecane).

Positive ion mass spectrometry for fragmentation of anthracene by low energy electron impact

SynopsisWe have studied the fragmentation of anthracene molecules induced by low energy electron impact. A reflectron time-of-flight mass spectrometer has been used to mass resolve and detect positively charged fragments. Using computer controlled data acquisition, mass spectra have been measured as a function of electron impact from 5 to 100 eV. The mass spectra show clear indications of double and triple ionization. Ion yield curves and appearance energies for most of the positive ions have been extracted from the data.

Positive ion mass spectrometry for fragmentation of 5-fluorouracil by low energy electron impact

SynopsisFragmentation of 5-fluorouracil in the gas phase has been studied using low energy electron impact. Positive ions have been mass resolved and detected using a reflectron time-of-flight mass spectrometer. Mass spectra have been measured as a function of electron impact from 5 to 100 eV, and ion yield curves and appearance energies for most of the positively charged fragments have been extracted.

Determination of warfarin and warfarin alcohols in dried blood spots by ultra-high performance liquid chromatography coupled to electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS)

A method for the simultaneous determination of warfarin and its active metabolites (warfarin alcohols) in dried blood spots samples was developed and validated. The procedure consisted of the collection of blood sample spots on Whatman 903 filter paper and the liquid extraction of the analytes by a 3:1v/v methanol/acetonitrile mixture from 6mm diameter disks punched out from dried blood spots. Extracted samples were analyzed by ultra-high performance liquid chromatography coupled to electrospray positive ionization and tandem mass spectrometry. Chromatographic separation was carried out in isocratic condition at 25°C on a Poroshell 120 EC-C18 reversed phase column with a mobile phase consisting of 65% water containing 0.1% (v/v) formic acid and 35% acetonitrile containing 0.1% (v/v) formic acid at a flow rate of 0.5mLmin−1.The method did not show any detectable interference or matrix effect. The limits of detection were 0.007, 0.006 and 0.004ng/mL for RR/SS-warfarin alcohols, RS/SR-warfarin alcohols and warfarin, respectively. Warfarin and warfarin alcohols recovery from dried blood spots sample was almost quantitative (range 96–103%). The intra- and inter-day precision (expressed as relative standard deviation) was always below 10%. Time stability studies highlighted that the concentrations of warfarin and both diastereoisomers of warfarin alcohols in a pooled blood sample were stable for at least 1month at −20°C and after 4cycles of thaw–freeze. Moreover, warfarin, RR/SS- and RS/SR-warfarin alcohols concentrations in the dried blood spots were stable for at least 7days at 25°C.Hematocrit value had a minor influence on the analytical result compared to blood drop volume. The method was successfully applied to the analysis of 15 blood samples from patients undergoing warfarin therapy and demonstrated its suitability as an alternative for warfarin and warfarin alcohols measurement in plasma. Good correlations between concentrations in plasma and dried blood spots were demonstrated for all three analytes, with correlation coefficients r≥0.95. Warfarin concentration in dried blood spot correlated well with warfarin dosage (r=0.70, p<0.01) but not with international normalized ratio (r=0.47, p=0.0784) when all the enrolled patients (n=15) were considered. However, this correlation increased (r=0.80, p=0.0302) if a single patient was monitored over time.These preliminary results highlighted that the developed method can provide useful information for the therapeutic drug monitoring of warfarin and its active metabolites, although further studies in a larger clinical trial are needed.

SiNx coatings deposited by reactive high power impulse magnetron sputtering: Process parameters influencing the residual coating stress

The residual coating stress and its control is of key importance for the performance and reliability of silicon nitride (SiNx) coatings for biomedical applications. This study explores the most important deposition process parameters to tailor the residual coating stress and hence improve the adhesion of SiNx coatings deposited by reactive high power impulse magnetron sputtering (rHiPIMS). Reactive sputter deposition and plasma characterization were conducted in an industrial deposition chamber equipped with pure Si targets in N2/Ar ambient. Reactive HiPIMS processes using N2-to-Ar flow ratios of 0 and 0.28–0.3 were studied with time averaged positive ion mass spectrometry. The coatings were deposited to thicknesses of 2 μm on Si(001) and to 5 μm on polished CoCrMo disks. The residual stress of the X-ray amorphous coatings was determined from the curvature of the Si substrates as obtained by X-ray diffraction. The coatings were further characterized by X-ray photoelectron spectroscopy, scanning electron microscopy, and nanoindentation in order to study their elemental composition, morphology, and hardness, respectively. The adhesion of the 5 μm thick coatings deposited on CoCrMo disks was assessed using the Rockwell C test. The deposition of SiNx coatings by rHiPIMS using N2-to-Ar flow ratios of 0.28 yield dense and hard SiNx coatings with Si/N ratios &amp;lt;1. The compressive residual stress of up to 2.1 GPa can be reduced to 0.2 GPa using a comparatively high deposition pressure of 600 mPa, substrate temperatures below 200 °C, low pulse energies of &amp;lt;2.5 Ws, and moderate negative bias voltages of up to 100 V. These process parameters resulted in excellent coating adhesion (ISO 0, HF1) and a low surface roughness of 14 nm for coatings deposited on CoCrMo.

Development and validation of a selective and effective pressurized liquid extraction followed by liquid chromatography–mass spectrometry method for the determination of fructosazine analogues in the ammonia treated extract of Eugenia jambolana Lamarck seeds

This study describes a selective and effective pressurized liquid extraction (PLE) coupled with HPLC-DAD-ESI/MS method for the identification and quantification of three fructosazine analogues (FZAs), fructosazine, 2,6- and 2,5-deoxyfructosazine in Madeglucyl® (MG) which is an ammonia treated extract of Eugenia jambolana Lamarck seeds, and is the world’s first anti-diabetic phytodrug. FZAs were extracted from MG by PLE using methanol as extraction solvent. The PLE extract was then analyzed directly by HPLC-DAD-ESI/MS without cleanup step. Chromatographic separation of these highly related structures was achieved on a porous graphic carbon (PGC) column. The identification of the target FZAs was confirmed by the similar retention time, similar UV and MS spectra to the corresponding pure standards. The quantification was performed by using an electrospray positive ionization mass spectrometry in the selected ion monitoring (SIM) mode. The PLE procedure was optimized and overall method was validated in terms of sensitivity, linearity, selectivity and matrix effect, precision, accuracy and recovery, and stability of the target FZAs in the aqueous solution and in the PLE extracts solution of MG. The developed method was proved to be selective, sensitive, precise, accurate for the quantification of FZAs in MG.

Mass spectrometry of positive ions in capacitively coupled low pressure RF discharges in oxygen with water impurities

A capacitively coupled RF oxygen discharge is studied by means of mass spectroscopy. Mass spectra of neutral and positive species are measured in the mid plane between the electrodes at different distances between plasma and mass-spectrometer orifice. In the case of positive ions, as expected, the largest flux originates from . However, a significant number of impurities are detected, especially for low input powers and larger distances. The most abundant positive ions (besides ) are , , and . In particular, for the case of hydrated hydronium ions (n = 1, 2) a surprisingly large flux (for low pressure plasma conditions) is detected. Another interesting fact concerns the ions. Despite the relatively high ammount of water impurities ions are present only in traces. The reaction mechanisms leading to the production of the observed ions, especially the hydrated hydronium ions are discussed.

Evaluation of the Effects of Ketoconazole and Voriconazole on the Pharmacokinetics of Oxcarbazepine and Its Main Metabolite MHD in Rats by UPLC-MS-MS.

Oxcarbazepine (OXC), a second-generation antiepileptic drug, undergoes rapid reduction with formation of the active metabolite 10,11-dihydro-10-hydroxy-carbazepine (MHD) in vivo. In this study, a method for simultaneous determination of OXC and MHD in rat plasma using ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS-MS) was developed and validated. Under given chromatographic conditions, OXC, MHD and internal standard diazepam were separated well and quantified by electrospray positive ionization mass spectrometry in the multiple reaction monitoring transitions mode. The method validation demonstrated good linearity over the range of 10-2,000 ng/mL for OXC and 5-1,000 ng/mL for MHD. The lower limit of quantification was 5 ng/mL for OXC and 2.5 ng/mL for MHD, respectively. The method was successfully applied to the evaluation of the pharmacokinetics of OXC and MHD in rats, with or without pretreatment by ketoconazole (KET) and voriconazole (VOR). Statistics indicated that KET and VOR significantly affected the disposition of OXC and MHD in vivo, whereas VOR predominantly interfered with the disposition of MHD. This method is suitable for pharmacokinetic study in small animals.

Thermally evaporated (oxide) iron on an alumina barrier layer by ToF-SIMS

The authors report the positive and negative ion time-of-flight secondary ion mass spectrometry spectra using Bi32+ primary ions at 50 keV of an Fe film (6 nm) that had been thermally evaporated on a thin film of alumina (ca. 35 nm) on a silicon wafer. This surface had been exposed to the air; it had previously been shown by x-ray photoemission spectroscopy (XPS) to be entirely oxidized. The positive ion secondary ion mass spectrometry spectrum shows Fe+, FeH+, and FexOyHz+ species. The negative ion spectrum shows FeOH−, FexOyHz−, and OH− species.

160 keV 26Al-AMS with a single-stage accelerator mass spectrometer

Proof-of-principle 26Al-AMS analysis is achieved with a single-stage accelerator mass spectrometer (SSAMS) utilising very low ion energy. The SSAMS operates by discriminating against atomic isobar interference in a negative ion source and suppressing molecules with thick gas stripper. Resulting 1+ ions counting is with a surface barrier detector. The NEC designed SSAMS for 14C analysis is a popular model accelerator mass spectrometer and the developed further capability might be a significant addition to established 26Al-AMS capacity. Measurements at these energies should also be sufficient for alternative 26Al positive-ion mass spectrometry (PIMS).

Application of Hydrophilic Interaction Liquid Chromatography Combined with Positive and Negative Ionization Mass Spectrometry for the Analysis of PSP Toxins

Hydrophilic interaction chromatography (HILIC) is a useful technique for the separation of PSP toxins that offers a good selectivity and increases the sensitivity of their detection by mass spectrometry (MS/MS). In this study the analytical conditions of the separation and detection were optimized by testing the effect of some components as column, the organic modifier (type and percentage)in the mobile phase, pH, buffer character and concentration and flow rate. Both types of ionization (positive and negative) were used in the same run. The best results were obtained with a ZIC-HILIC (Merck), using a gradient elution, detecting the toxins in negative ionization mode during the first part of the chromatogram, and in positive mode during the final part. The limits of detection (LODs) and quantization (LOQs) obtained were acceptable for all toxins and low enough for routine monitoring, as well of the analysis time. The matrix effects were observed for some PSP toxins in mussel extracts, so a simple solid phase extraction (SPE) method to clean up the extracts prior the analysis was developed.

Structural Characterization of γ‐Terpinene Thin Films Using Mass Spectroscopy and X‐Ray Photoelectron Spectroscopy

Understanding the polymerization mechanism of a precursor is indispensable to enhance the requisite material properties. In situ mass spectroscopy and X‐ray photoelectron spectroscopy is used in this study to understand the RF plasma polymerization of γ‐terpinene. High‐resolution mass spectra positive ion mass spectrometry data of the plasma phase demonstrates the presence of oligomeric species of the type [M+H]+ and [2M+H]+, where M represents a unit of the starting material. In addition, there is abundant fragmented species, with most dominant being [M+] (136 m/z), C10H13+ (133 m/z), C9H11+ (119 m/z), and C7H9+ (93 m/z). The results reported in this manuscript enables to comprehend the relationship between the degree of incorporation of oxygen and the rate of deposition with the input RF power.

Determination of N-nitrosodiethanolamine, NDELA in cosmetic ingredients and products by mixed mode solid phase extraction and UPLC–tandem mass spectrometry with porous graphitic carbon column through systemic sample pre-cleanup procedure

A rapid, sensitive, accurate and specific ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) method for the detection of N-nitrosodiethanolamine (NDELA), a highly toxic contaminant in cosmetic raw materials and products was developed and validated. Systematized sample preparation steps were developed according to product types. Various SPE cartridges and columns were examined to establish the condition of SPE and chromatographic separation for NDELA. Sample cleanup steps consisting of solvent and liquid-liquid extraction tailored to the various sample matrix types were established prior to mixed mode SPE (Bond Elut AccuCAT). Chromatographic separation was achieved within 7 min on a porous graphitic carbon (PGC) column using a gradient elution with the mobile phase of 1mM ammonium acetate containing 0.1% acetic acid and methanol. NDELA was monitored using an electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode (m/z 134.9>103.7(quantifier) and 73.7(qualifier ion)) with d8-NDELA (m/z 143.1>111.0) as internal standard. The standard curves were linear over the concentration range of 1-100 ng/mL with a correlation coefficient higher than 0.99. The limit of detection (LOD) and the limit of quantification (LOQ) was 10 and 20 μg/kg, respectively (0.5 and 1 ng/mL in standard solution). The intra- and inter-day precisions were estimated to be below 11.1% and accuracies were within the range of 90.8-115.8%. The validated method was successfully applied to the analysis of real samples including raw materials, skin care, make-up, shampoos and hair products.

The analysis of azocyclotin and cyhexatin residues in fruits using ultrahigh-performance liquid chromatography-tandem mass spectrometry

Ultrahigh-performance liquid chromatography coupled to electrospray positive ionization and tandem mass spectrometry (UHPLC-ESI(+)-MS/MS) was applied for the quantification and identification of azocyclotin and cyhexatin in fruit samples.

Characterization of the prepolymer and gel of biocompatible poly(xylitol sebacate) in comparison with poly(glycerol sebacate) using a combination of mass spectrometry and nuclear magnetic resonance

AbstractAtmospheric pressure chemical ionization (negative ion) and electrospray ionization (positive and negative ion) mass spectrometry and nuclear magnetic resonance (1H NMR and 13C NMR) were used to characterize the prepolymer (i.e. uncrosslinked) poly(xylitol sebacate) (PXS), prepared by condensation of xylitol [X] and sebacic acid [S], and this was compared with poly(glycerol sebacate) (PGS), prepared by condensation of glycerol [G] and [S]. The PXS prepolymer, prepared at 118 °C/24 h, gave predominantly 1‐acyl‐[XS] and 1,5‐diacyl‐[SXS] species and smaller amounts of 2‐acyl substitutions. At 130 °C/24 h an increase of the amount of the 1‐acyl and 1,5‐diacyl substitutions in the [(SX)n], [S(XS)n] and [(XS)nX] series was identified, as was found for the analogous PGS prepolymer ([(SG)n], [S(GS)n] and [(GS)nG] series). Novel tetrahydrofuranyl species were found in the PXS prepolymer. Further polymerization of the PXS prepolymer gave a gel which when analysed using 13C NMR was shown to mainly contain 1‐acyl and 1,5‐diacyl substitutions. © 2014 Society of Chemical Industry

Quick and simple LC-MS/MS method for the determination of simvastatin in human plasma: application to pharmacokinetics and bioequivalence studies

A simple, rapid, and sensitive method based on liquid chromatography-tandem mass spectrometry for the quantitative determination of simvastatin in human plasma was developed and validated. After a simple extraction with methyl tert-butyl ether, the analyte and internal standard (lovastatin) were analyzed using reverse-phase liquid chromatography, on a Kinetex C18column (100 × 4.6 mm, 2.6 μm) using acetonitrile: ammonium acetate (2 mM + 0.025 % formic acid) (70: 30, v/v) as a mobile phase in a run time of 3.5 min. Detection was carried out using electrospray positive ionization mass spectrometry in the multiple-reaction monitoring mode. The method was linear over 0.04-40.0 ng/mL concentration range. The mean extraction recovery of simvastatin was 82% (RSD within 15%). Intraday and interday precisions (as relative standard deviation) were all ≤8,7% with accuracy (as relative error) of ±8%. This rapid and reliable method was successfully applied for a bioequivalence study of 40 mg of simvastatin orally disintegrating tablets in 44 healthy volunteers, showing that this method is suitable for the quantification of simvastatin in human plasma samples for pharmacokinetics and bioequivalence studies.