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.

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.

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.

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.

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.

Continuous-flow size-based separation of microparticles by microchip electromagnetophoresis.

A novel microchip separation method for microparticles based on electromagnetophoresis was developed. In this method, a double Y-shape microchip with two inlets and two outlets was used for particle separation. Microparticles of two different sizes were suspended in an aqueous solution and introduced into the microchannel from one inlet with a stream of the aqueous solution without particles from another inlet by a hydrodynamic flow. Then, the particles with two different sizes were migrated orthogonal to the flow by elctromagnetophoresis and separated to different outlets based on the difference in their electromagnetophoric velocities, which depended on the size of particle. Using this method, at one outlet, 89% of 3 µm polystyrene latex particles were isolated from a mixture of 3 and 10 µm particles. By combining hydrodynamic focusing, both 3 and 10 µm particles were recovered at the different outlets with 100% purity and recovery, respectively. Even the separation of 3 and 6 µm particles was also achieved with about 98% recovery and purity. Moreover, yeast cells and polystyrene particles could be separated with high separation efficiency by this technique.

Electromagnetophoretic migration velocity of organic microdroplets with surfactants using permanent magnets.

By using the electromagnetophoretic migration technique with permanent magnets, the electromagnetophoretic migration velocimetry in a droplet-based system was demonstrated for organic droplets dispersed in an aqueous solution. Migration of 2-fluorotoluene droplets with a diameter of 8 - 16 µm dispersed in 1.0 M KCl aqueous solution could be achieved in the same manner as for solid particles. The effect of cetyltrimethylammonium bromide (CTAB) on the electromagnetophoretic migration velocity of the droplets was also investigated. When the concentration of CTAB was in the range from 10(-6) to 10(-4) M, the electromagnetophoretic migration velocity of 2-fluorotoluene droplets decreased as the concentration of CTAB increased. With this measurement, we could successfully characterize organic droplets in terms of the amount of adsorbing surfactants by evaluating the surface conductivity of organic droplets calculated based on their electromagnetophoretic migration velocity.

Dynamic electromagnetophoretic force analysis of a single binding interaction between lectin and mannan polysaccharide on yeast cell surface.

Using the electromagnetophoretic buoyancy for a microparticle in a silica capillary containing an electrolyte solution, the dynamic force dissociation kinetics of the interaction between the mannan polysaccharide on a yeast cell surface and lectins bound to the silica capillary wall have been studied by using the two different modes of the increasing force mode and the constant force mode. Lectins used in the study were concanavalin A (Con A), Hippeastrum hybrid lectin (HHL), Galanthus nivalis lectin (GNL) and Narcissus pseudonarcissus lectin (NPL). The dynamic force measurement by the two different modes gave the spontaneous dissociation rate constant, k(off), of the polysaccharide-lectin binding and the critical increment of bond length at the transition state, Deltax. It was found that the value of k(off) for Con A was the smallest among the lectins studied, due to the strongest binding interaction. It was also confirmed that the magnetophoretic pulling force decreased the transition free energy just like an enzyme.

Electromagnetophoretic force measurement of a single binding interaction between lectin and yeast cell surfaces.

A novel measurement method of the binding force between a micrometer-sized particle and a solid surface in an electrolyte solution has been established by using the electromagnetophoretic buoyancy on the particle. By this method, we investigated the binding force between a yeast cell surface and an oligosaccharide-binding protein, concanavalin A (Con A), fixed on a silica capillary wall. The force measurement was carried out up to 60 pN. In a lower surface concentration of Con A, yeast cells could be desorbed by a force less than 60 pN. However, in a higher surface concentration after treated by 1 mg ml(-1) solution, yeast cells were adsorbed with a force stronger than 60 pN. In this case, the addition of 10 mg ml(-1) D-mannose solution to the medium reduced the binding force to less than 60 pN. The observed adsorption force of yeast cells ranged within 30 - 40 pN, regardless of the interfacial amount of Con A. This force was thought to be the single binding force between a mannose group of the cell surface and an active site of Con A. Moreover, the dissociation rate constant of the single binding of yeast cell and Con A complex was determined as 4.6 x 10(-3) s(-1) and the increment of the binding distance at the transition state as 0.33 nm from the desorption kinetic experiments of yeast cell under the constant pulling conditions of 10, 20 and 30 pN. Such satisfactory results demonstrate the novel advantages of the present method.

New principle of electromagnetophoretic adsorption-desorption microchromatography.

A new principle for the chromatographic micro-separation of micrometer-sized particles in liquid has been invented by switching the electromagnetophoretic force in a capillary flow system. The principle is the combination of the Stokes force by the bulk flow and the adsorption-desorption force on a capillary inner surface controlled by an electromagnetophoretic buoyancy generated by an alternative current and a homogenous magnetic field. The observed retention profiles of test micro-particles was explained by the "zigzag" migration model mainly depended on particle size and their adsorption force to the capillary wall. By this method, we could succeed to separate polystyrene particles of 10 microm and 20 microm in diameter dispersed in 1 M KCl solution containing 0.01% Triton X-100 using only 1 mm long fused-silica capillary under 10T.

High-magnetic-field electromagnetophoresis of micro-particles in a capillary flow system.

The electromagnetophoretic migration of micro-particles in a capillary flow system was demonstrated using a homogeneous magnetic field applied at right angles to an electric current. We utilized a high-magnetic-field of 10 T for observing this phenomenon. When the direction of the electric current was alternatively changed, polystyrene latex particles in a flowing aqueous medium migrated zigzag affected by a Lorentz force exerted on the medium. Carbon particles also migrated in the same manner with polystyrene particles. Further, we tried the electromagnetophoretic migration of biological particles, such as yeasts and human red blood cells. The migration velocity component perpendicular to the flow was proportional to both the electric current and the magnetic flux density. These results proved that the dominant force of the zigzag migration was an electromagnetophoretic buoyancy generated in the flowing medium. Moreover, it was found that the force exerted on the particles in the magnetic field of 10 T was sufficient for the desorption of particles adsorbed on the capillary wall.

Migration analysis of micro-particles in liquids using microscopically designed external fields.

The recent development of new migration methods of micro-particles in liquids using various external fields is reviewed. The combination of a laser scattering force and a photothermal effect produced photothermal-conversion laser-photophoresis. A dielectric field generated in a planer or a capillary quadrupole electrode realized dielectrophoresis. Using a micrometer-scaled magnetic field gradient, the "Magnetophoretic velocimetry" of micro-particles was invented. Furthermore, the Lorentz force generated by combining an electric field and a magnetic field was utilized for electromagnetophoresis. These new methods were overlooked and the advantages in analytical use were discussed.

Magnetophoresis and electromagnetophoresis of microparticles in liquids.

The magnetic field-induced migration of particles in liquids is a highly-promising technique for the micro-separation analysis of bioparticles, such as cells and large DNA. Here, new methods that make use of magnetophoresis and electromagnetophoresis to induce the migration of microparticles in liquids are briefly reviewed. Magnetic force and Lorentz force are utilized in the new methods. Some typical examples of the use of these methods are described, and the advantages of using a superconducting magnet for them are demonstrated.

Simultaneous measurement of the migration velocity and adsorption force of micro-particles using an electromagnetophoretic force under a high magnetic field.

A simultaneous measurement technique for determining the migration velocity of a micrometer-sized particle in a capillary and the adsorption force to the inner surface of the capillary has been proposed. This technique is based on an electromagnetophoretic force being exerted on a micro-particle in an electrolyte solution, which is governed mainly by the electromagnetic buoyancy, when a homogeneous magnetic field is applied at a right angle to the electric current through the medium. By the electromagnetic buoyancy, micro-particles such as polystyrene, carbon and yeast were migrated perpendicular to the direction of the electric current and reached a fused-silica wall. A switching of the current direction could desorb the particle from the wall, and allowed to calculate the detaching force from the desorbing current. The migration velocity normalized to the size in the magnetic field of 10 T was increased in the order of yeast, carbon and polystyrene, while reflecting the decreasing order of the apparent conductivity of the particles. The desorption force could be measured up to 1 nN with a sensitivity of pN. The observed interaction forces of polystyrene and carbon were in the range of 250-600 pN with large deviations.