Takuya Shimosaka

Fabrication of polyester microchannels and their applications to capillary electrophoresis

Inexpensive and disposable polyester microchips were fabricated through photolithographic and wet-chemical etching procedure, followed by replication using an imprinting method at room temperature. Laboratory-scale laser-induced fluorescence equipment was employed as a detection system. The generation of electroosmotic flow (EOF) on the polyester channels was discussed in this paper. Surfactants in the running buffer had a significant effect on the EOF depending on their types. The epsilon potential of the electric double layer formed by adsorbing sodium lauryl sulfate molecules on the wall of polyester channels seemed to be constant within the buffer pH investigated. EOF could also be suppressed to zero by adding polyoxyethylene 23 lauryl ether into the running buffer. The separation of two laser dyes was obtained using polyester chips through both micellar electrokinetic chromatography and capillary zone electrophoresis. The polyester channels modified with 10-undecen-1-ol exhibited a dramatically high-separation efficiency compared with the conventional fused-silica capillary tubes.

A flow-based enzyme-linked immunosorbent assay on a polydimethylsiloxane microchip for the rapid determination of immunoglobulin A.

A flow-based enzyme-linked immunosorbent assay (ELISA) on a polydimethylsiloxane (PDMS) microchip has been developed for the rapid determination of immunoglobulin A (IgA). The analytical principle of this integrated method is the same as the conventional sandwich-type ELISA. A primary antibody (anti-IgA) was adsorbed on the surface of a PDMS microchannel, and then an antigen (IgA) and a secondary antibody (anti-IgA HRP labeled) were reacted successively. The resulting antigen-antibody complex, fixed on the surface of the microchannel, was detected using Amplex((R)) Red and a fluorescent imaging system. The calibration curve of the IgA standard solution was linear in the range of 0-50ng/mL at the flow rate of 10muL/min. This flow rate corresponds to the reaction time of 4.8s. Compared to the conventional assay on a 96-well microtiter plate, the present assay on the microchip dramatically shortened the reaction time necessary for the enzyme-substrate reaction from 30min to 4.8s, i.e., to 1/375. The amounts of the reagent and sample were also reduced to 1/100 compared to the 96-well microtiter plate.

Final report on international key comparison CCQM-K53: Oxygen in nitrogen

Gravimetry is used as the primary method for the preparation of primary standard gas mixtures in most national metrology institutes, and it requires the combined abilities of purity assessment, weighing technique and analytical skills. At the CCQM GAWG meeting in October 2005, it was agreed that KRISS should coordinate a key comparison, CCQM-K53, on the gravimetric preparation of gas, at a level of 100 ?mol/mol of oxygen in nitrogen. KRISS compared the gravimetric value of each cylinder with an analytical instrument. A preparation for oxygen gas standard mixture requires particular care to be accurate, because oxygen is a major component of the atmosphere. Key issues for this comparison are related to (1) the gravimetric technique which needs at least two steps for dilution, (2) oxygen impurity in nitrogen, and (3) argon impurity in nitrogen. The key comparison reference value is obtained from the linear regression line (with origin) of a selected set of participants. The KCRV subset, except one, agree with each other. The standard deviation of the x-residuals of this group (which consists of NMIJ, VSL, NIST, NPL, BAM, KRISS and CENAM) is 0.056 ?mol/mol and consistent with the uncertainties given to their standard mixtures. The standard deviation of the residuals of all participating laboratory is 0.182 ?mol/mol. With respect to impurity analysis, overall argon amounts of the cylinders are in the region of about 3 ?mol/mol; however; four cylinders showed an argon amount fraction over 10 ?mol/mol. Two of these are inconsistent with the KCRV subset. The explicit separation between two peaks of oxygen and argon in the GC chromatogram is essential to maintain analytical capability. Additionally oxygen impurity analysis in nitrogen is indispensable to ensure the preparative capability. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

International comparison CCQM-K82: methane in air at ambient level (1800 to 2200) nmol/mol

The CCQM-K82 comparison was designed to evaluate the degrees of equivalence of NMI capabilities for methane in air primary reference mixtures in the range (1800 to 2200) nmol/mol. The balance gas for the standards was either scrubbed dry real air or synthetic air. CH4 in air standards have been produced by a number of laboratories for many years, with more recent developments focused on standards at atmospheric measurement concentrations and aimed at obtaining agreement between independently produced standards. A comparison of the differences in primary gas standards for methane in air was previously performed in 2003 (CCQM-P41 Greenhouse gases. 1 and 2) with a standard deviation of results around the reference value of 30 nmol/mol and 10 nmol/mol for a more limited set of standards. This can be contrasted with the level of agreement required from field laboratories routinely measuring atmospheric methane levels, set by Data Quality Objectives (DQO) established by the World Meteorological Organization (WMO) to reflect the scientifically desirable level of compatibility for CH4 measurements at the global scale, currently set at 2 nmol/mol (1 sigma). The measurements of this key comparison took place from May 2012 to June 2012. Eight laboratories took part in this comparison coordinated by the BIPM and NIST. Key comparison reference values were calculated based on Cavity Ring Down Spectroscopy Measurements performed at the BIPM, combined with participants gravimetric values to identify a consistent set of standards. Regression analysis allowed predicted values for each standard to be calculated which acted as the KCRVs. In this comparison reported standard uncertainties by participants ranged from 0.50 nmol/mol to 2.4 nmol/mol and the uncertainties of individual KCRVs ranged from 0.68 nmol/mol to 0.71 nmol/mol. The standard deviation of the ensemble of standards about the KCRV value was 1.70 nmol/mol. This represents a greater than tenfold improvement in the level of compatibility of methane in air standards compared to that demonstrated in 2003. Further improvements in the compatibility of standards will require improved methods and uncertainties for the measurement of trace level methane in balance gases. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Evaluation of lateral resolution of scanning surface microscopy by total internal reflection with thermal lens effect

We have developed a novel method for in situ and non-destructive surface analyses, or a total internal reflection with thermal lens spectroscopy (TIR-TLS), which has sufficient sensitivity to monitor phenomena in thin films, such as lipid bilayers. In this study, we applied TIR-TLS to microscopy for surface analyses, and we experimentally obtained its lateral resolution using the edge of a chromium film made by a photolithography technique. The obtained resolution was 20 microm, which was 60% of the diameter of an excitation beam at the interface. The estimated resolution with a simple model agreed with the experimental one, and from this model, TIR-TLS microscopy has the same resolution as that of ordinary optical microscopy. The microscopy by TIR-TLS was applied to a sample whose contrast was too weak to be visually seen, and an image of the sample was obtained without any loss of resolution.

High-precision GC-TCD for verification of gravimetrically prepared primary gas standards of oxygen in nitrogen

Gravimetric preparation method is one of the most precise and accurate methods for preparing primary standard gas mixtures (PSMs). Although the principle and execution of the gravimetric method are not complicated, the uncertainty of the gravimetric method is so small that a seemingly minor error can result in an incorrect amount fraction of the PSM. Therefore, verifying the amount fraction of the PSMs is a necessary step. We perform a high-precision comparison between the PSMs of oxygen in nitrogen (∼100 µmol per mol) using a well-stabilized gas chromatograph with a thermal conductivity detector (GC-TCD) system in conjunction with high-precision pressure monitoring for sample amount correction and a quality-control cylinder technique in which the PSMs were analyzed against a reference cylinder, which leads to the cancellation of the drift in the sensitivity of the GC-TCD. After optimization of the measurement conditions, the relative expanded uncertainty reaches 0.022% (coverage factor k = 2), which is equivalent to the uncertainty of the PSMs. The Consultative Committee for Amount of Substance of the Comité International des Poids et Mesures performed a key comparison CCQM-K53, in which one of the PSMs used in this work was compared and agreed well with PSMs prepared by other National Metrology Institutes. These results show that the GC-TCD system developed in this work is sufficiently reliable, precise and accurate to verify PSMs.

Application and Validation of a Determination Method Using Post-column Reaction Gas Chromatography of Nitrogen-containing Organic Compounds

In this work, we applied post-column reaction gas chromatography (GC) using a flame ionization detector (FID) system to study nitrogen-containing organic compounds (NOCs). The results were subsequently validated. After separation by column, the target components were converted to carbon dioxide using an oxidizing catalyst and then reduced to methane, followed by detection using an FID. SI-traceable testing mixtures containing NOCs (isoprocarb, napropamide, and pendimethalin) were prepared by the gravimetric blending method. These mixtures were analyzed using a post-column reaction GC-FID system; standard materials of hydrocarbons were used as calibrants in this analysis. The determined values were compared with the values obtained for samples prepared at the corresponding concentrations, and statistical analyses were performed in all cases. It was shown that the determined and prepared values agreed well with each other within the uncertainty limits.

Development of an Analytical System Based on a Magneto-pneumatic Oxygen Analyzer for Atmospheric Oxygen Determination

A high-precision analytical system to observe the variations in the amount fractions of atmospheric oxygen with a very small uncertainty was developed. The system comprises a magneto-pneumatic oxygen analyzer and three automatic pressure controllers. The drift of the analyzers signal intensity can be reduced when the amount fractions of oxygen in the sample and reference gases are similar because the temperature coefficient of the analyzer linearly depends on the difference between these amount fractions. The repeatability of oxygen determination and the long-term stability of the system were tested to assess the applicability of the analyzer to field-based measurements for continuous atmospheric observations. The standard deviation of the average for 10-min measurements in the 5-day long-term stability test was 0.7 μmol mol-1 after a temperature correction. This indicates that the system can continuously measure the amount fractions of oxygen in the atmosphere for a few days without interruption for any calibration and/or compensation for the signal drift.

Accurate determination of formaldehyde amount fraction in cylinders using mixtures of primary standards of formaldehyde in nitrogen prepared by a gravimetric permeation method

A reliable method for the quantification of formaldehyde (HCHO) amount fractions is required, because the results of comparisons between individual techniques used for the quantification of HCHO amount fractions often disagree owing to unknown causes. One reliable method comprises the quantification of HCHO using working gas standards in which the HCHO amount fraction is linked to the International System of Units (SI). In this work, the SI traceability of HCHO amount fractions in mixtures of commercial HCHO in nitrogen in cylinders used as working gas standards was established by calibrating them against primary gas standard mixtures (mixtures of HCHO in N2), which were prepared by a permeation method and were traceable to SI. The expanded relative uncertainties (k = 2) of the amount fractions in the primary gas standard mixtures were less than 0.6 %. The quantitative values of the amount fractions in the working gas standards had relative expanded uncertainties (k = 2) of 0.7 %–0.9 %.

Final report of international comparison APMP.QM-S2.2015 of oxygen in nitrogen at 0.2 mol/mol

1) National Metrology Institute of Japan, National Institute of Advanced Industry and Science Technology, Umezono 1-1-1, Tsukuba, Ibaraki, Japan 2) Center for Measurement Standards, Industrial Technology Research Institute, Kuang Fu Rd, Hsinchu, 30011, Taiwan 3) National Institute of Metrology Thailand, 3⁄4-5 Moo 3, Klong 5, Klong Luang, Pathumthani 12120, Thailand 4) Chemicals Evaluation and Research Institute, Japan, 1600 Shimotakano, Sugito-machi, Kitakatsushika-gun, Saitama, Japan 5) Karaganda branch of RSE, Kazakhstan Institute of Metrology, Angerskaya st., 22/2, 100009, Kazakhstan