Characterization of balsam sulfide via pyrolysis-gas chromatography-mass spectrometry/sulfur chemiluminescence detection and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Balsam sulfide, produced by the reaction of turpentine/rosin and sulfur, has been used as one of the raw materials of liquid gold to decorate ceramics and tableware with thin gold film for more than 100 years. The characterization of balsam sulfide is still insufficient because of its compositional complexity. In this study, balsam sulfide was characterized using pyrolysis-gas chromatography (Py-GC)-mass spectrometry (MS) and Py-GC with sulfur chemiluminescence detection (SCD) as well as matrix-assisted laser desorption/ionization time-of-flight MS (MALDI-TOFMS). Py-GC-MS/SCD analyses of balsam sulfide and its model samples revealed that the low molecular weight reaction products were mainly composed of compounds of one α-pinene unit reacted with 1-3 sulfur atoms. In the analysis of the high molecular weight components by MALDI-TOFMS, the products of two or three α-pinene units crosslinked by sulfur atoms were observed. It was found that dehydrogenation reaction proceeded gradually with the increase in the reaction time.

Control of One-Handed Helicity in Polyacetylenes: Impact of an Extremely Small Amount of Chiral Substituents.

Controlling the one-handed helicity in synthetic polymers is crucial for developing helical polymer-based advanced chiral materials. We now report that an extremely small amount of chiral biphenylylacetylene (BPA) monomers (ca. 0.3-0.5 mol %) allows complete control of the one-handed helicity throughout the polymer chains mostly composed of achiral BPAs. Chiral substituents introduced at the 2-position of the biphenyl units of BPA positioned in the vicinity of the polymer backbones contribute to a significant amplification of the helical bias, as interpreted by theoretical modeling and simulation. The helical structures, such as the helical pitch and absolute helical handedness (right- or left-handed helix) of the one-handed helical copolymers, were unambiguously determined by high-resolution atomic force microscopy combined with X-ray diffraction. The exceptionally strong helix-biasing power of the chiral BPA provides a highly durable and practically useful chiral material for the separation of enantiomers in chromatography by copolymerization of an achiral functional BPA with a small amount of the chiral BPA (0.5 mol %) due to the robust helical scaffold of the one-handed helical copolymer.

Analysis of polyisoprene oligomers via in situ silver nanoparticle formation on thin-layer chromatography plate using matrix-assisted laser-induced desorption/ionization mass spectrometry.

The direct mass spectrometry (MS) detection of polyisoprene (PI) oligomers on a thin-layer chromatography (TLC) plate using matrix-assisted laser-induced desorption/ionization (MALDI) with silver trifluoroacetate as the cationization reagent was investigated. The spots of PI oligomers and silver trifluoroacetate on the TLC plate resulted in brown materials after UV laser irradiation. It was suggested that silver trifluoroacetate yielded Ag nanoparticles as brown materials after heating via laser irradiation. The nanoparticles behaved as an inorganic matrix and a source of Ag+ adduct in the analysis of PI oligomers. The use of organic matrices together with silver trifluoroacetate reduced the signal intensity of PI oligomers on MALDI-MS on a TLC plate. The separation of PI oligomers (polymerization degree, n = 5-11) by TLC resulted in a single elliptical spot without a clear separation after the chromatographic procedure. However, in MS imaging, differences in migration lengths based on degrees of polymerization were clearly observed and the degrees of polymerization were identified without comparison with standards.

Characterization of styrene/methyl methacrylate/n-butyl acrylate terpolymer with Kendrick mass defect analysis in electrospray ionization-ion mobility spectrometry-mass spectrometry.

RATIONALE: Synthesis of copolymer is one of the major approaches to establish sophisticated polymers. In copolymer analysis in mass spectrometry (MS), an increase in the number of monomer unit species and/or the polymer molecular weight results in complex mass spectra, and a method is required to solve this problem. METHODS: In this study, electrospray ionization-ion mobility spectrometry-mass spectrometry was employed to analyze styrene/methyl methacrylate/n-butyl acrylate (St/MMA/nBA) terpolymers via Kendrick mass defect (KMD) analysis. An expanded Fourier transform-based noise reduction method for the copolymer was developed to improve the decrease in the signal/noise ratio observed in the higher m/z region. RESULTS: Low-mass terpolymers (0.7 kDa) were identified by comparing the observed and theoretical results using a two-dimensional KMD plot. For the 1.5 kDa terpolymer, the signal overlap, of which KMD value was not matched with the theoretical one, was interpreted by the shift from the theoretical value in the KMD plot. For the 3.0 kDa terpolymers, the compositional candidates were determined by the prediction based on the compositional information of low-mass terpolymers previously analyzed. The 4.5 kDa terpolymer was interpreted after the expanded noise filtering. CONCLUSIONS: The terpolymers, whose molecular weight was up to 4.5 kDa, were successfully characterized at a molecular level. The dependency of the St/MMA/nBA composition on molecular weight was observed; that is, the nBA content decreased with an increase in the molecular weight.

Pyrolysis-GC-MS analysis of ingested polystyrene microsphere content in individual Daphnia magna.

Microplastic (MP) pollution in the aquatic environment is a cause for increasing concern. However, analyzing MPs ingested by small organisms, such as zooplankton, is difficult because of the low content and small size of the ingested MPs. We attempted to determine the content of ingested MPs in individual zooplankton using pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). To establish zooplankton model of MP ingestion, individual Daphnia magna were cultivated separately in microplate cells with polystyrene (PS) microspheres (10 µm in diameter, 245,000 particles, 135 µg) under different conditions. To prepare calibration curves for determining ingested PS content, approximately 100-150 µg of commercially available Daphnia-based powdered fish food, roughly corresponding to the weight of a single D. magna organism, was mixed with PS microspheres (0.005-26 µg) and analyzed using Py-GC-MS at 600 °C. In the resulting pyrograms, peaks of the styrene monomer and trimer from PS were detected, and linear relationships were obtained between the relative peak area and the amount of added PS. Finally, the cultivated zooplankton were individually subjected to Py-GC-MS analysis, and the ingested PS content in each zooplankton was successfully determined. Individual zooplankton cultured with PS in the absence of food ingested 2.3-7.9 µg of PS particles, whereas that in the presence of food (Chlorella vulgaris) ingested only 0.1-0.2 µg of PS particles. This result suggests that zooplankton might preferentially ingest ordinary food when both food and MPs are present, although further systematic studies are necessary to validate this observation.

End group analysis of poly(methylmethacrylate)s using the most abundant peak in electrospray ionization-ion mobility spectrometry-tandem mass spectrometry and Fourier transform-based noise filtering.

RATIONALE: We recently developed the characterization method for synthetic polymers weighing more than a few tens of kilodalton using electrospray ionization-ion mobility spectrometry-tandem mass spectrometry, in which the m/z value of the most abundant peak was used for characterization. However, the identification of the most abundant peak from the isotopic peaks was often difficult due to the background noise. METHODS: Here, we employed a noise reduction method using Fourier transform (FT) filtering. In the power spectrum obtained using FT of the mass spectrum of the multiple charged analytes, the significant signals in the low-frequency region and at frequency z are observed for the analytes of z charges. When the signals in both regions were used for inversed FT (i.e., the signals in other regions were zero padded), a noise-filtered mass spectrum was obtained. RESULTS: In the analysis of poly(methylmethacrylate)s weighing 13-17 kDa, mass spectra without noise filtering with relatively high-intensity noise (than signal) were complicated to identify the most abundant peak. On the contrary, the most abundant peak was clearly identified from the mass spectra after FT-based noise filtering, and end group composition was estimated successfully. CONCLUSIONS: The proposed FT-based noise filtering for the mass spectrum is effective to characterize multiply charged synthetic polymers weighing more than a few tens of kilodalton using electrospray ionization-ion mobility spectrometry-tandem mass spectrometry.

Characterizing chain-end structures formed during initiation reactions of radical polymerization for MMA-St-BA terpolymer using pyrolysis-gas chromatography/atmospheric pressure chemical ionization high-resolution time-of-flight mass spectrometry.

RATIONALE: Analyzing polymer end groups using pyrolysis (Py) gas chromatography/mass spectrometry (GC/MS) in multi-component polymer samples is not an easy task because of the insufficient sensitivity, selectivity, and mass resolution of conventional Py-GC/MS systems. METHODS: A new Py-GC/MS system using an atmospheric pressure chemical ionization (APCI) source combined with high-resolution time-of-flight mass spectrometry (TOFMS) was used for end-group analysis of a methyl methacrylate (MMA)-styrene (St)-butyl acrylate (BA) terpolymer (P (MMA-St-BA)), which was radically polymerized using 2,2'-azobis(2-methylbutyronitrile) (AMBN) as an initiator. RESULTS: Five possible pyrolyzates, comprising an AMBN fragment and a monomer unit, formed during the initiation reactions from one of the three types of end groups, were selectively detected and exclusively identified in their respective extracted ion chromatograms for molecule-related ions, such as M+ and [M + H]+ , with a narrow mass window of ±2 milli m/z units. CONCLUSIONS: It was demonstrated that Py-APCI-TOFMS is a powerful technique to characterize in detail the complex end groups in multi-component polymer samples, because of the soft ionization nature of APCI and the high mass resolution of TOFMS.

Electrospray ionization-ion mobility spectrometry-high resolution tandem mass spectrometry with collision-induced charge stripping for the analysis of highly multiply charged intact polymers.

Polymers with large molecular weight are difficult to interpret using electrospray ionization-mass spectrometry (ESI-MS) due to the generation of various highly multiply charged analytes. Although ESI-ion mobility spectrometry (IMS)-MS is effective in reducing the complexity of the mass spectrum, this approach is insufficient for analyzing highly multiply charged polymers. In this study, we propose a method combining tandem mass spectrometry (quadrupole and high-resolution time-of-flight MS, QMS/TOFMS), IMS, and collision-induced charge stripping (CICS) for analyzing large intact polymers (∼40 kDa), which are highly multiply charged. The number of charges can be estimated from a Fourier transform power spectrum of a mass spectrum when the charge number is less than approximately 20. Interpretations of the spectra of poly(ethylene oxide)s (PEOs) weighing 20 kDa, poly(methyl methacrylate)s weighing 22 kDa, and methoxy-PEO-maleimide weighing 40 kDa were successfully demonstrated with isotope level and polymerization degree level separations, respectively. In the proposed method, a mixture can be analyzed for relatively small (a few kDa) and large (a few tens of kDa) polymers simultaneously without any sample pretreatment.

Frequency division multiplex HPLC-MS for simultaneous analyses.

Multiplex high-performance liquid chromatograph-mass spectrometry (HPLC-MS), in which multiple HPLCs and one MS are hyphenated, is an approach for high throughput analysis in HPLC-MS. A general multiplex HPLC-MS method employs a column-switching technology, and only one HPLC is connected to one MS at a time. In the present study, we propose a novel multiplex HPLC-MS system for simultaneous HPLC-MS analyses. In this study, multiple HPLCs are hyphenated with one MS without a column-switching mechanism, and a mixed-chromatogram is observed by the MS. Here, we employ a frequency division multiplexing (FDM) technique used in communication engineering to extract any chromatogram from the mixed-chromatogram. When a modulator (chopper or ion-gate type) is set between each ion source and the MS, each modulator blocks each sample stream with an individual frequency. In theory, each chromatogram can be extracted from the mixed-chromatogram via a signal processing based on a Fourier transform (FT), frequency-based signal extraction, and reversed FT. In the actual experiment, two HPLCs are hyphenated with one MS (2HPLC-1MS). The use of chopper type modulators leads to the extraction and restoration of each chromatogram from the mixed-chromatogram. However, each restored-chromatogram involves signal interference. On the other hand, the ion-gate modulation system successfully resulted in restored-chromatograms without interference. The potential of the novel multiplex HPLC-MS system based on FDM is confirmed with respect to the simultaneous and continuous analyses of plural samples.

Analysis of Multiply Charged Poly(ethylene oxide-co-propylene oxide) Using Electrospray Ionization-Ion Mobility Spectrometry-Mass Spectrometry.

Poly(ethylene oxide), poly(propylene oxide), and their copolymer (poly(EO-co-PO)) were analyzed by electrospray ionization-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS). ESI produced multiply charged analytes of 2 to 5 Na+ additions, and they were separately observed in a 2D map of m/z value vs. drift time. The collision cross-section of the analyte polymers was almost linearly proportional to (molecular weight)0.644, except for the analytes with 2Na+ addition; a nonlinear relation called "folding" was significantly observed for the analytes with 2Na+ addition. An increase in electrostatic repulsion, because of the increase in Na+ addition, suppressed the folding of the polymer. Analyses of poly(EO-co-PO) with different EO compositions revealed that the copolymer with high EO composition tended to show folding. The separation of highly multiply charged poly(EO-co-PO)s with different EO contents by ESI-IMS-MS was successfully demonstrated.

On the Kendrick Mass Defect Plots of Multiply Charged Polymer Ions: Splits, Misalignments, and How to Correct Them.

The Kendrick mass defect (KMD) analysis of multiply charged polymeric distributions has recently revealed a surprising isotopic split in their KMD plots-namely a 1/z difference between KMDs of isotopes of an oligomer at charge state z. Relying on the KMD analysis of actual and simulated distributions of poly(ethylene oxide) (PEO), the isotopic split is mathematically accounted for and found to go with an isotopic misalignment in certain cases. It is demonstrated that the divisibility (resp. indivisibility) of the nominal mass of the repeating unit (R) by z is the condition for homolog ions to line up horizontally (resp. misaligned obliquely) in a KMD plot. Computing KMDs using a fractional base unit R/z eventually corrects the misalignments for the associated charge state while using the least common multiple of all the charge states as the divisor realigns all the points at once. The isotopic split itself can be removed by using either a new charge-dependent KMD plot compatible with any fractional base unit or the remainders of KM (RKM) recently developed for low-resolution data all found to be linked in a unified theory. These original applications of the fractional base units and the RKM plots are of importance theoretically to satisfy the basics of a mass defect analysis and practically for a correct data handling of single stage and tandem mass spectra of multiply charged homo- and copolymers. Graphical Abstract ᅟ.

On-line Analysis of Catalytic Reaction Products Using a High-Pressure Tandem Micro-reactor GC/MS.

When a GC/MS system is coupled with a pressurized reactor, the separation efficiency and the retention time are directly affected by the reactor pressure. To keep the GC column flow rate constant irrespective of the reaction pressure, a restrictor capillary tube and an open split interface are attached between the GC injection port and the head of a GC separation column. The capability of the attached modules is demonstrated for the on-line GC/MS analysis of catalytic reaction products of a bio-oil model sample (guaiacol), produced under a pressure of 1 to 3 MPa.

High-resolution Kendrick Mass Defect Analysis of Poly(ethylene oxide)-based Non-ionic Surfactants and Their Degradation Products.

Matrix assisted laser desorption ionization (MALDI) high-resolution mass spectrometry (HRMS) and the recently introduced high-resolution Kendrick mass defect (HRKMD) analysis are combined to thoroughly characterize non-ionic surfactants made of a poly(ethylene oxide) (PEO) core capped by esters of fatty acids. A PEO monostearate surfactant is first analyzed as a proof of principle of the HRKMD analysis conducted with a fraction of EO as the base unit (EO/X with X being an integer) in lieu of EO for a regular KMD analysis. Data visualization is greatly enhanced and the distributions detected in the MALDI mass spectrum are assigned to a pristine (H, OH)-PEO as well as mono- and di-esterified PEO chains with palmitate and stearate end-groups in HRKMD plots computed with EO/45. The MALDI-HRMS/HRKMD analysis is then successfully applied to the more complex case of ethoxylated hydrogenated castor oil (EHCO) found to contain a large number of hydrogenated ricinoleate moieties (up to 14) in its HRKMD plot computed with EO/43, departing from the expected triglyceride structure. The exhaustiveness of the MALDI-HRMS/HRKMD strategy is validated by comparing the so-obtained fingerprints with results from alternative techniques (electrospray ionization MS, size exclusion and liquid adsorption chromatography, ion mobility spectrometry). Finally, aged non-ionic surfactants formed upon hydrolytic degradation are analyzed by MALDI-HRMS/HRKMD to easily assign the degradation products and infer the associated degradation routes. In addition to the hydrolysis of the ester groups observed for EHCO, chain scissions and new polar end-groups are observed in the HRKMD plot of PEO monostearate arising from a competitive oxidative ageing.

Multi-imaging of Cytokinin and Abscisic Acid on the Roots of Rice (Oryza sativa) Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

Plant hormones act as important signaling molecules that regulate responses to abiotic stress as well as plant growth and development. Because their concentrations of hormones control the physiological responses in the target tissue, it is important to know the distributions and concentrations in the tissues. However, it is difficult to determine the hormone concentration on the plant tissue as a result of the limitations of conventional methods. Here, we report the first multi-imaging of two plant hormones, one of cytokinin [i.e., trans-zeatin (tZ)] and abscisic acid (ABA) using a new technology, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) imaging. Protonated signals of tZ (m/z 220.1) and ABA (m/z 265.3) were chosen on longitudinal sections of rice roots for MS imaging. tZ was broadly distributed about 40 mm behind the root apex but was barely detectable at the apex, whereas ABA was mainly detected at the root apex. Multi-imaging using MALDI-TOF-MS enabled the visualization of the localization and quantification of plant hormones. Thus, this tool is applicable to a wide range of plant species growing under various environmental conditions.

Retention in high-performance liquid chromatography at -196°C.

Recently, we have developed ultralow-temperature high-performance liquid chromatography (HPLC) at -196°C using liquid nitrogen-based mobile phases. In this study, a retention model for ultralow-temperature HPLC, in which adsorption exchange and "pseudo partition" modes are combined, is proposed to describe the effect of the mobile phase composition on the retention of analytes. The experimental results agreed well with the equation of the proposed model. It was revealed that, in the low and high additive concentration regions, adsorption exchange and pseudo partition retention, respectively, dominate the analyte retention. The difference in the retention behavior between bare-silica and octadecylsilyl-modified silica (ODS) columns was also studied. Retention of alkanes in the ODS column was greater than that in the bare-silica one. Addition of both ethane and ethylene to the mobile phase in the ODS column afforded the elution of propylene, which was not eluted with the bare-silica column at -196°C.

Effect of surfactant species and electrophoretic medium composition on the electrophoretic behavior of neutral and water-insoluble linear synthetic polymers in nonaqueous capillary zone electrophoresis.

We have recently demonstrated the separation of neutral and water-insoluble linear synthetic polymers in nonaqueous capillary zone electrophoresis (NACZE) using a cationic surfactant of cetyltrimethylammonium chloride (CTAC). In this study, eight ionic surfactants were investigated for the separation of four synthetic polymers (polystyrene, polymethylmethacrylates, polybutadiene, and polycarbonate); only three surfactants (CTAC, dimethyldioctadecylammonium bromide, and sodium dodecylsulfate) caused their separation. The order of the interaction between the polymers and the surfactants depended on both the surfactant species and the composition of the electrophoretic medium. Their investigation revealed that the separation is majorly affected by the hydrophobic interactions between the polymers and the ionic surfactants. In addition, the electrophoretic behavior of polycarbonate suggested that electrostatic interaction also affects the selectivity of the polymers.

High Performance Liquid Chromatography at -196 °C.

Ultralow temperature high-performance liquid chromatography (HPLC) was developed using a liquefied gas as the mobile phase. HPLC separation of low molecular weight alkanes at -196 °C with liquid nitrogen mobile phase was successfully achieved, whereas their GC separation at -196 °C using helium gas mobile phase failed to elute the analytes due to strong adsorption. Prior to the further study of HPLC at -196 °C, the effect of column temperature on the chromatographic behavior was investigated, and it was found that the retention of analytes drastically increased when the column temperature was over the boiling point of the mobile phase. As the study of retention control in HPLC at -196 °C, the mobile phases of nitrogen and methane mixtures were investigated. The addition of methane to the nitrogen mobile phase suppressed the retention of the analytes (tetra-deuterated methane, ethane, and propane), that is, the retention on HPLC at ultralow temperature could be controlled by the mobile phase composition, akin to the typical retention in HPLC. The selectivity toward the n- and iso-alkane in HPLC at -196 °C was altered compared with that in GC separation at room temperature. A significant enhancement of retention of alkanes compared with alkanes were observed in HPLC at -196 °C.

Variation in the chromatographic, material, and chemical characteristics of methacrylate-based polymer monoliths during photoinitiated low-temperature polymerization.

Both the separation behavior and the structure of a polymer monolith column depends on both the reaction solution composition and the polymerization conditions. In photoinitiated low-temperature polymerization, polymerization temperature, irradiation intensity, and polymerization time were key factors to control the monolith characteristics. In this study, the effect of polymerization time on the chromatographic, material, and chemical characteristics of poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths was studied using pyrolysis-gas chromatography, Raman spectroscopy, inverse size exclusion chromatography, scanning electron microscopy, and chromatographic methods. Both butyl methacrylate and ethylene dimethacrylate monomers were incorporated into the monolith as the polymerization time increased, and it resulted in increases in both the flow resistance (decrease in both permeability and total/through pore porosities) and retention factors. The longer polymerization time led to lower relative amounts of free methacrylate functional groups in the monolith, i.e. cross-linking was enhanced. The increase of the polymerization time from 8 to 12 min significantly reduced the separation efficiency for the retained analyte, whereas an increase in the fraction of the mesoporosity was observed.

Hydrogenation Reactions during Pyrolysis-Gas Chromatography/Mass Spectrometry Analysis of Polymer Samples Using Hydrogen Carrier Gas.

Pyrolysis-gas chromatography/mass spectrometry of polymer samples is studied focusing on the effect of hydrogen (H2) carrier gas on chromatographic and spectral data. The pyrograms and the related mass spectra of high density polyethylene (HDPE), low density polyethylene, and polystyrene (PS) serve to illustrate the differences between the species formed in H2 and the helium environment. Differences in the pyrograms and the spectra are generally thought to be a result of the hydrogenation reaction of the pyrolyzates. From the peak intensity changes in the pyrograms of HDPE and PS, hydrogenation of unsaturated pyrolyzates is concluded to occur when the pyrolysis is done in H2. Moreover, additional hydrogenation of the pyrolyzates occurs in the electron ionization source of a MS detector when H2 is used as a carrier gas. Finally, the applicability of mass spectral libraries to characterize pyrograms obtained in H2 is illustrated using 24 polymers. The effect of the hydrogenation reaction on the library search results is found to be negligible for most polymer samples with polar and nonpolar monomer units.

Staggered-electromagnetophoresis with a Split-flow System for the Separation of Microparticles by a Hollow Fiber-embedded PDMS Microchip.

A novel microchip separation system for microparticles based on electromagnetophoresis (EMP) was developed. In this system, focusing and separation of flowing microparticles in a microchannel could be performed by staggered-EMP by controlling the electric current applied to the channel locally combined with the split-flow system for fractionation of eluates. To apply the electric current through the flushing medium in the microchannel, a hollow fiber-embedded microchip with multiple electrodes was fabricated. The hollow fiber was made by a semi-permeable membrane and could separate small molecules. This microchip allowed us to apply the electric current to a part of the microchannel without any pressure control device because a main channel contacted with the subchannels that had electrodes through the semi-permeable membrane. Moreover, the separation using this microchip was combined with the split-flow system at two outlets to improve separation efficiency. Using this system, with the split-flow ratio of 10:1, 87% of 3 µm polystyrene (PS) latex particles were isolated from a mixture of 3 and 10 µm particles. Even the separation of 6 and 10 µm PS particles was achieved with about 77% recovery and 100% purity. In addition, by controlling the applied current, size fractionation of polypropylene (PP) particles was demonstrated. Moreover, biological particles such as pollens could be separated with high separation efficiency by this technique.