Mass spectrometry最新文献

CO₃ ^-• and O₂ ^-• are known to be strong oxidizing reagents in biological systems. CO₃ ^-• in particular can cause serious damage to DNA and proteins by H^• abstraction reactions. However, H^• abstraction of CO₃ ^-• in the gas phase has not yet been reported. In this work we report on gas-phase ion/molecule reactions of CO₃ ^-• and O₂ ^-• with various molecules. CO₃ ^-• was generated by the corona discharge of an O₂ reagent gas using a cylindrical tube ion source. O₂ ^-• was generated by the application of a 15 kHz high frequency voltage to a sharp needle in ambient air at the threshold voltage for the appearance of an ion signal. In the reactions of CO₃ ^-•, a decrease in signal intensities of CO₃ ^-• accompanied by the simultaneous increase of that of HCO₃ ^- was observed when organic compounds with H-C bond energies lower than ∼100 kcal mol^-1 such as n-hexane, cyclohexane, methanol, ethanol, 1-propanol, 2-propanol, and toluene were introduced into the ion source. This clearly indicates the occurrence of H^• abstraction. O₂ ^-• abstracts H⁺ from acid molecules such as formic, acetic, trifluoroacetic, nitric and amino acids. Gas-phase CO₃ ^-• may play a role as a strong oxidizing reagent as it does in the condensed phase. The major discharge product CO₃ ^-• in addition to O₂ ^-•, O₃, and NO _x ^• that are formed in ambient air may cause damage to biological systems.

Temperature-resolved proton transfer reactions of multiply-protonated angiotensin I, disulfide-intact and -reduced lysozyme, and ubiquitin ions to primary, secondary and aromatic amines were examined in the gas phase. Absolute reaction rate constants for the proton transfer were determined from the intensities of the parent and product ions in mass spectra. Dramatic changes were observed in the distribution of product ions and the reaction rate constants. In particular, the rate constants for disulfide-intact lysozyme ions changed more drastically with the change in charge state and temperature compared to the corresponding values for disulfide-reduced ions. Proton transfer reactions were enhanced or suppressed as the result of the formation of complexes between the ions with gaseous molecules, which is related to changes in their conformation with changing.

We measured the Re/Os (¹⁸⁵Re/¹⁸⁸Os) and ¹⁸⁷Os/¹⁸⁸Os ratios from nanoparticles (NPs) using a multiple collector-inductively coupled plasma-mass spectrometer equipped with high-time resolution ion counters (HTR-MC-ICP-MS). Using the HTR-MC-ICP-MS system developed in this study, the simultaneous data acquisition of four isotopes was possible with a time resolution of up to 10 μs. This permits the quantitative analysis of four isotopes to be carried out from transient signals (e.g., <0.6 ms) emanating from the NPs. Iridium-Osmium NPs were produced from a naturally occurring Ir-Os alloy (ruthenosmiridium from Hokkaido, Japan; osmiridium from British Columbia, Canada; iridosmine from the Urals region of Russia) through a laser ablation technique, and the resulting nanoparticles were collected by bubbling water through a suspension. The ¹⁸⁷Os/¹⁸⁸Os ratios for individual NPs varied significantly, mainly due to the counting statistics of the ¹⁸⁷Os and ¹⁸⁸Os signals. Despite the large variation in the measured ratios, the resulting ¹⁸⁷Os/¹⁸⁸Os ratios for three Ir-Os bearing minerals, were 0.121±0.013 for Hokkaido, 0.110±0.012 for British Columbia, and 0.122±0.020 for the Urals, and these values were in agreement with the ratios obtained by the conventional laser ablation-MC-ICP-MS technique. The data obtained here provides a clear demonstration that the HTR-MC-ICP-MS technique can become a powerful tool for monitoring elemental and isotope ratios from NPs of multiple components.

The Kendrick analysis is used for the processing and visualization of mass spectra obtained from polymers containing C, H, O and/or Si with simple isotopic patterns (monoisotope=lightest isotope=most intense isotope for short chains). In the case of heteroatoms with complex isotopic patterns, the impact of the chosen isotope on point alignments in Kendrick plots has not been examined extensively. Rich isotopic patterns also make the evaluation of the mass and nature of the repeating unit and end-groups more difficult from the mass spectrum in the case of unknown samples due to the number of peaks and the absence of a monoisotopic peak. Using a polybrominated polycarbonate as running example, we report that horizontal point alignments can be obtained in a Kendrick plot using the mass of the most abundant isotope instead of the monoisotopic mass as is usually done. Rotating the plot ("reverse Kendrick analysis") helps to accurately evaluate the mass of the most abundant isotope of the repeating unit, as well as the nature of the brominated neutral expelled upon gentle heating (debromination or dehydrobromination). The whole procedure is then applied to the characterization of an unknown polybrominated flame retardant in an industrial formulation before and after heating.

Using a multi-turn time-of-flight (TOF) mass spectrometer, we have extracted a single xenon isotope ion, ¹²⁹Xe⁺, from its orbit at given a lap number without disturbing the rest of isotopes. After detecting the ¹²⁹Xe⁺ at 20 laps, the rest of the xenon isotope spectrum was obtained at 30 laps, which generated a TOF spectrum where the TOF difference between ¹²⁹Xe⁺ and ¹³⁰Xe⁺ was 87.4 μs while ¹³⁰Xe⁺ and ¹³¹Xe⁺ were 1.03 μs. The time distance between ¹²⁹Xe⁺ and other isotopes can be set by any lap difference that is a factor of 8.7 μs, which depends on the acceleration voltage and the mass of the ion. Method accuracy was verified by comparing the isotopic abundance ratio of the xenon sample after withdrawing one of the ions from the isotope cluster to the abundance ratio obtained from the conventional method. The TOF stability was also evaluated at various lap numbers between 10 to 230.

Combined therapy using photodynamic therapy (PDT) and chemotherapy has been proposed for anticancer-drug-resistant cancer cells. To evaluate the efficacy of such a combined therapy, the uptakes of an anticancer drug and a photosensitizer in cancer cells must be assessed. Mass spectrometry using matrix-assisted laser desorption/ionization can detect multiple drugs simultaneously. Human prostate cancer cells PC-3 or docetaxel-resistant cancer cells PC-3-DR were incubated in a serum-free medium containing a photosensitizer, protoporphyrin IX (PpIX), and an anticancer drug, docetaxel. A zeolite matrix was created by mixing 6-aza-2-thiothymine and NaY5.6 zeolite, and dissolving in water with 50% acetone. Ions were obtained with a time-of-flight mass spectrometer using a Nd:YAG laser at a wavelength of 355 nm. The cell morphology was preserved by washing the cells with ammonium acetate and drying in a vacuum after drug administration. Protonated PpIX (m/z 563.3) and the sodium adduct ion of docetaxel (m/z 829.9) were obtained from PC-3 cells simultaneously using the zeolite matrix. On the other hand, PpIX was detected but ions originating from docetaxel were not detected from PC-3-DR cells. The result indicated the efficacy of PDT for docetaxel-resistant cancer cells.

Protein phosphorylation mediated by protein kinases is one of the most significant posttranslational modifications in many biological events. The function and physiological substrates of specific protein kinases, which are highly associated with known signal transduction elements or therapeutic targets, have been extensively studied using various approaches; however, most protein kinases have not yet been characterized. In recent decades, many techniques have been developed for the identification of in vitro and physiological substrates of protein kinases. In this review, I summarize recent studies profiling the characteristics of kinases using mass spectrometry-based proteomics, focusing on the large-scale identification of in vitro substrates of the human kinome using a quantitative phosphoproteomics approach.

The rapid development of next-generation sequencing techniques has enabled single-cell genomic and transcriptomic analyses, which have revealed the importance of heterogeneity in biological systems. However, analytical methods to accurately identify and quantify comprehensive metabolites from single mammalian cells with a typical diameter of 10-20 μm are still in the process of development. The aim of this study was to develop a single-cell metabolomic analytical system based on highly sensitive nano-liquid chromatography tandem mass spectrometry (nano-LC-MS/MS) with multiple reaction monitoring. A packed nano-LC column (3-μm particle-size pentafluorophenylpropyl Discovery HSF5 of dimensions 100 μm i.d.×180 mm) was prepared using a slurry technique. The optimized nano-LC-MS/MS method showed 3-132-fold (average value, 26-fold) greater sensitivity than semimicro-LC-MS/MS, and the detection limits for several hydrophilic metabolites, including amino acids and nucleic acid related metabolites were in the sub-fmol range. By combining live single-cell sampling and nano-LC-MS/MS, we successfully detected 18 relatively abundant hydrophilic metabolites (16 amino acids and 2 nucleic acid related metabolites) from single HeLa cells (n=22). Based on single-cell metabolic profiles, the 22 HeLa cells were classified into three distinct subclasses, suggesting differences in metabolic function in cultured HeLa cell populations. Our single-cell metabolomic analytical system represents a potentially useful tool for in-depth studies focused on cell metabolism and heterogeneity.

The compounds inside rice koji have been thoroughly investigated as an essential material in making many food-related products, including sake. However, these studies focused only on quantitative aspects, leaving features that can still be uncovered if seen from a new perspective. Visualization of the metabolites inside rice koji may as well be the new angle needed to retrieve more information regarding rice koji making. Here we utilized mass spectrometry imaging (MSI) to visualize the distribution of sugars, sugar alcohols, and amino acids inside rice koji. Imaging results revealed that several sugars alcohols and amino acids were shown to have characteristic distribution near the edges or surface of rice koji. Furthermore, the distribution appears to be correlated with the different structure of rice koji. This study is the first report of using MSI to visualize sugars, sugar alcohols, and amino acids in rice koji.