fabrication of electrochemical /photoelectrochemical biosensor using novel...

INV-01 47

ASIANALYSIS XIII 8–11 December 2016, Chiang Mai, THAILAND

Fabrication of electrochemical /photoelectrochemical biosensor

using novel nano materials A. Gopalan,2,3, N. Muthuchamy,1, K-P. Lee1,2,3,*

1Department of Chemistry Education, Kyungpook National University, Republic of Korea, 2Research Institute of Advanced Energy Technology, Kyungpook National University, Republic of Korea,

3Department of Nanoscience and Nanotechnology, Kyungpook National University,

Republic of Korea. E-mail: [email protected]

Sensors based on electrochemical methods posses advantages such as portability, low cost, short analysis

time and excellent sensitivity Also, photoelectrochemical sensor is a new kind of developing analytical

device based on the photoelectrochemical properties of materials. Because of its remarkable

sensitivity, inherent miniaturization, portability and easy integration, photoelectrochemical analysis is

becoming a promising analytical technique. In recent years, carbon nanomaterials such as carbon

nanotubes(CNT), graphene(G), and nanodiamonds (ND), plasmonic nanoparticles such as gold (Au)

nanostructures etc. have been extensively utilized, in their pristine and functionalized forms, toward

sensor applications. Importantly, these nanomaterials can be suitably modified to generate sensing

probes for few of the analytes. This lecture focuses our research activities on the development of few

novel nanomaterials towards fabrication of electrochemical/photeoelectrochemical sensors.(1-4)

Polyaniline chains were grafted onto nanodiamond (PANI-g-ND) through electrochemical

polymerization of aniline in the presence of amine functionalized ND. A robust and effective composite

film comprising PANI-g-ND/gold particles was subsequently prepared. Cyclic voltammograms revealed

that PANI-g-ND/Au/cyt c exhibited an excellent electrocatalysis towards the detection of nitrite ions.

Differential pulse voltammetry of PANI-g-ND/Au/cyt c revealed a wide linear concentration range (0.5

μM-3 mM) for current responses, sensitivity (88.2 μA/mM) and low detection limit (0.16 μM) towards

the electrochemical detection of nitrite ions. We demonstrate the use of a novel CP-based MCNB,

comprising a functionalized CP, Fc, and Cu NP for efficient NEG sensing. We have chosen poly-

(aniline-co-anthranilic acid)-grafted G sheets [designated as G-PANI(COOH)] as the parent CP, and

branched polyethyleneimine (b-PEI) as the cross-linking polymer, ferrocene carboxyaldehyde (Fc-CHO)

as the cross-linker, and Cu NPs for the preparation of the new MCNB. The components G-

PANI(COOH), b-PEI, and Fc-CHO were integrated to form a cross-linked network structure, and further

dispersed with the Cu NPs to obtain G-PANI(COOH)-PEI-Fc/Cu-MCNB. Electrochemical

measurements showed that the G-PANI(COOH)-PEI-Fc/Cu-MCNB/GCE modified electrode exhibited

good electrocatalytic behavior towards the detection of glucose in a wide linear range (0.50 to 15 mM),

with a low detection limit (0.16 mM) and high sensitivity (14.3 µA mM−1 cm−2).

We reported the fabrication of a novel electrochemical-PEC dual mode biosensor utilizing the beneficial

components such as G, TiO2(G)-NWs and CS and transforming them into a NS through establishment of

chemical interactions between modified TiO2-NWs and G. We designated the assemblies of G and

TiO2(G)-NWs as G/Ti(G)-3DNS. The G/Ti(G)-3DNS was further modified with CS and used for

immobilization of ChOx to fabricate the biosensor G/Ti(G)-3DNS/CS/ChOx and used as the

electrochemical-PEC dual mode biosensor for the detection of cholesterol. The G/Ti(G) 3DNS/CS/ChOx

bioelectrode was selective to cholesterol with a remarkable sensitivity (3.82 μA/cm2 mM) and a lower

detection limit (6 μM). Also, G/Ti(G) 3DNS/CS/ChOx functioned as photoelectrode and exhibited

selective detection of cholesterol under a low bias voltage and light irradiation.

We are pursuing continuous research for exploring the various kind of nanomaterials for sensing

application.

Keywords: Sensors; Electrochemical; Phoelectrochemical; Nanomaterials

References

1) A.I. Gopalan, N. Muthuchamy, S. Komathi, K-P Lee, Biosensors and

Bioelectronics, Volume 84, 53-63 (2016)

2) A.I. Gopalan, N. Muthuchamy, S. Komathi, K-P Lee, Biosensors and Bioelectronics,

doi:10.1016/j.bios.2016.07.017 (2016)

3) S. Komathi, N. Muthuchamy, K-P. Lee, A-I. Gopalan, Biosensors and Bioelectronics, 84,

64-71(2016)

4) A. I. Gopalan, S.Komathi, G. Sai Anand, K-P. Lee, Biosensors and Biolectronics, 46, 36-

141(2013)

mailto:[email protected]

http://dx.doi.org/10.1016/j.bios.2016.07.017

INV-02 48


Environmental applications of sequential injection vapor generation

coupled with chemiluminescence detection and mass spectrometry

Kei Toda1*

1Department of Chemistry, Faculty of Science, Kumamoto University, Kumamoto 860-8555, Japan

*E-mail: [email protected]

Vapor generation is advantageous to make sensitivity much higher than conventional direct

absorbance or fluorescence detection, because large amount of analyte is introduced to the detection

system via the vapor generation. Sequential injection analysis (SIA) is a convenient way to convert

the analytes volatile and to make vaporization. In this presentation, basics of vapor generation is

discussed first and then applications to selenium detection in power plant wastewater and to

investigation of chemical dynamics at the seawater/atmosphere interface are presented.

Vaporization is advantageous as described before but applicability is limited by volatility of the

analytes. We demonstrated that compounds having Henry’s law constant (KH) less than 1.0 can be

applied to the vapor generation analysis method [1]. Vaporization rate (dmol/dt in mol/min) is

expressed as eq. 1 using analyte initial concentration Caq0 (nM), sample volume V (L), KH (M/atm),

bubbling carrier airflow rate F (L/min) and bubbling time t (min) [2]. From eq. 1, maximum vapor

concentration Cv max (ppbv) is obtained at t = 0, and expressed as eq. 2 where we can see the

sensitivity is determined only by KH and independent from the vaporization conditions such as

bubbling flow rate or water sample volume. However the time needed for completing vaporization

can be shortened by higher air flow rate and smaller sample volume for a certain compound.

The KH values of heavy metal hydrides are much smaller than 1.0 so that they are appropriate

compounds for vapor generation. In addition, these hydride vapors react with gaseous O3 to produce

strong chemiluminescence. Since 3.11 disaster in 2011, old thermal power plants were re-driven to

recover the Japanese electricity. However coal, used for the fuel, contains selenium (Se) and the Se

content in the power plant wastewater became another issue [3]. To monitor Se in wastewater,

novel system was used which comprised of sequential injection system coupled with a heated

reactor, cooled vapor generator, and chemiluminescence (CL) detector. Whereas generated H2Se

produced CL in the reaction with O3, CL for AsH3 was much higher and arsenic strongly interfered

the Se analysis. To solve this problem we investigated the vaporization carefully and succeeded to

make vapors of As and Se separately. These heavy metals were discriminatively determined every

30 min.

Volatile sulfur compounds govern the climate and atmospheric environment, especially dimethyl

sulfide (DMS) is the dominant. Dimethylsulfoniopropionate (DMSP) is the source of DMS, which

is produced in phytoplankton for controlling osmotic pressure. We developed sequential

vaporization method for DMS and DMSP separately and vaporized DMS was detected by CL [4] or

IMR-MS [2]. DMS and DMSP in seawater and atmospheric DMS were monitored for a week and it

was found that flux of DMS from seawater surface dramatically varied diurnally with change in

physical and biological environment parameters. [1] K. Toda, H. Kuwahara, S. Ohira, On-site measurement of trace-level sulfide in natural waters by vapor

generation and microchannel collection. Environ. Sci. Technol., 45, 5622–5628 (2011).

[2] S. Iyadomi, K. Ezoe, S. Ohira, K. Toda, Monitoring variations of dimethyl sulfide and

dimethylsulfoniopropionate in seawater and the atmosphere based on sequential vapor generation and ion

molecule reaction mass spectrometry. Environ. Sci.: Processes Impacts, 18, 464–472 (2016).

[3] K. Ezoe, S. Ohyama, M.A. Hashem, S. Ohira, K. Toda, Automated determinations of selenium in thermal

power plant wastewater by sequential hydride generation and chemiluminescence detection. Talanta, 148,

609–616 (2016).

[4] T. Nagahata, H. Kajiwara, S. Ohira, K. Toda, Simple field device for measurement of dimethyl sulfide and

dimethylsulfoniopropionate in natural waters, based on vapor generation and chemiluminescence

detection. Anal. Chem., 85, 4461−4467 (2013).

Keywords: Sequential vaporization; Arsenic and selenium; Dimethyl sulfide (DMS) and

dimethylsulfoniopropionate (DMSP); gas-phase chemiluminescence detection; ion molecule reaction

mass spectrometry (IMR-MS)


INV-03 49


Office equipment, inkjet printing and chemically functional inks – a toolbox for

(microfluidic) paper-based analytical devices (µPADs)

Daniel Citterio*

Department of Applied Chemistry, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Japan


(Microfluidic) paper-based analytical devices, commonly referred to as (µ)PADs, have evolved into

promising tools for on-site analysis. The fact that (µ)PADs are very low cost, normally self-

standing devices not requiring external instrumentation and reagents, easy to operate without

intensive training, and safely disposable by incineration, makes them of high interest for application

in resource-limited settings, such as for example encountered in home-healthcare, clinical

diagnostics in remote areas of developing countries, and field analysis. In addition, they can also be

useful to reduce costs and simplify analytical assays in standard laboratory environments.

From the beginning, our research group has recognized the potential of inkjet printing technology, a

method allowing the deposition of picoliter-sized droplets with high spatial resolution and

precision, in the fabrication of paper-based analytical devices.1-8 In fact, devices as simple as

common office inkjet printers can be used to both microfluidically pattern filter paper substrates and

to deposit a broad variety of chemically functional inks required for analytical assays. Depending

on the physical characteristics of a chemically functional ink (e.g. viscosity, surface tension,

presence of particulate matter), either a thermally-actuated or a piezoelectrically-actuated printer is

used. Only in the case where the use of organic solvents with high dissolving power is required (e.g.

deposition of solutions of hydrophobic polymers), a more specialized material inkjet printer has to

be applied. In addition to inkjet printers, we are making use of other standard office equipment,

including wax printers and hot laminators.

During this presentation, an overview of our more recent work will be given. This includes

miniaturized µPADs suitable for the analysis of sub-microliter samples,9 ion-selective optical

sensing µPADs for alkali metal cations with integrated pH-buffering system, as well as fully inkjet-

printed sodium and potassium ion-selective electrode (ISE) systems including both working and

reference electrodes requiring no conditioning.

References:

1. Abe, K.; Suzuki, K.; Citterio, D. Anal. Chem. 2008, 80, 6928-6934.

2. Abe, K.; Kotera, K.; Suzuki, K.; Citterio, D. Anal. Bioanal. Chem. 2010, 398, 885-893.

3. Komuro, N.; Takaki, S.; Suzuki, K.; Citterio, D. Anal. Bioanal. Chem. 2013, 405, 5785-5805.

4. Maejima, K.; Tomikawa, S.; Suzuki, K.; Citterio, D. RSC Adv. 2013, 3, 9258-9263.

5. Soga, T.; Jimbo, Y.; Suzuki, K.; Citterio, D. Anal. Chem. 2013, 85, 8973-8978.

6. Yamada, K.; Takaki, S.; Komuro, N.; Suzuki, K.; Citterio, D. Analyst 2014, 139, 1637-1643.

7. Yamada, K.; Henares, T. G.; Suzuki, K.; Citterio, D. Angew. Chem. Int. Ed. 2015, 54, 5294-5310.

8. Yamada, K.; Henares, T. G.; Suzuki, K.; Citterio, D. ACS Appl. Mater. Interfaces 2015, 7,

24864-24875.

9. Tenda, K.; Ota, R.; Yamada, K.; Henares, T.; Suzuki, K.; Citterio, D. Micromachines 2016, 7, 80.

Keywords Inkjet printing; Ion-selective optodes; Ion-selective electrodes; Colorimetry

INV-04 50


Development of Software for Polymer Product Evaluation using TG/PI-QMS with

Skimmer Interface.

Takahisa Tsugoshi1*, Yuji Mishima2

1National Metrology Institute of Japan, National Institute of Advanced Industrial Science and Technology,

Tsukuba, 4305-8563, Japan 2Kobe Material Testing Laboratory Co., Ltd., Taito-ku, Tokyo 110-0015, Japan


Evolved gas analyses with pyrolysis were carried out for some kinds of polymer products using a

newly developed apparatus of Skimmer interface-connected [TG/DTA]/[Photo-Ionization-QMS]

that is “ThermoMass Photo” by Rigaku corporation. The apparatus provides an advantage of real-

time monitoring of pyrolyzed species because of no transformation and no retention time of the

evolved species using the skimmer interface. The PI technique is fragmentless ionization that is one

of the soft ionization techniques offering no fragmentation during ionization, so that one peak

indicates one chemical species reflecting its molecular weight. In the mass spectrum, the evolved

species can be distinguished without any separation techniques, such as gas chromatography. The

combination of the fragmentless ionization mass spectrometry and TG/DTA offers the

discrimination of polymers with the same chemical structure and different thermal properties.

For the fragmentless mass spectra, there are some minor peaks due to small amount of additives,

and the small peaks may be ignored or designated as noise. For this purpose, multivariate analysis

such as “Residual Analysis” is effective to identify these minor peaks due to small amount species.

In case of usual apparatus such as capillary-connected TG/MS and Py-GC/MS, unstable species

evolved by pyrolysis must be adsorbed or transformed on the inner wall of transfer device for

evolved gaseous species. Hence it follows that detection of these species is impossible.

The software is newly developed, and useful for polymer discrimination by spectrum matching and

product comparison by residual analysis. DCR (Different Calculation method of Regression

analysis) offers the spectrum matching, so that discrimination between not only homo-polymers but

blended polymers using homo-polymer database only, without database of blended polymers.

The software-assisted evaluation has been carried out to shock-resisted polystyrene that is blended

polymer of polystyrene and styrene-butadiene copolymer. The result indicated PS:SB = 78 %:22 %,

and discriminated that SB was ABA block copolymer with 28 wt% styrene. It is appropriate to the

product specification.

Keywords evolved gas analysis; fragmentless ionization; skimmer interface; multivariate analysis;

polymer

INV-05 51


DNA Interaction Probed by Evanescent Wave Cavity Ring-down Absorption

Spectroscopy and C-dots Optical Characterization

King-Chuen Lin Department of Chemistry, National Taiwan University, Taipei, and Institute of Atomic and Molecular Sciences,

Academia Sinica, Taipei 106, Taiwan

E-mail: [email protected], web site: http://ntuchemkclin.weebly.com/research-interest.html

Evanescent wave cavity ring-down absorption spectroscopy (EW-CRDS) is employed to study

interaction and binding kinetics of DNA strands by using gold nanoparticles (Au NPs) as sensitive

reporters. These Au NPs are connected to target DNA of study that hybridizes with the complementary

DNA fixed on the silica surface. By the absorbance of Au NPs, the interaction between two DNA

strands may be examined. The binding efficiency that is affected by such as ion concentration, buffer

pH and temperature is also examined. This approach is then applied to the label-free detection of the

DNA mutation diseases using the sandwich hybridization assay. The EW-CRDS method appears to

have great potential for the investigation of the kinetics of a wide range of biological reactions.[1 and

Fig.1] Steady-state and time-resolved fluorescence spectroscopy techniques were used to probe multi-

fluorescence resulting from citric acid derived carbon-dots (C-dots). Commonly, both carboxyl/amine

functionalized C-dots exhibit three different emissive states corresponding to the carbon-core and

surface domain. The shorter wavelength fluorescence (below 400 nm) originates from carbon-core

absorption band at 290 nm, whereas the fluorescence (above 400 nm) is caused by two surface states at

350 and 385 nm. In addition to three emissive states, a molecular state was also found in amine

functionalized C-dots. Time-resolved emission spectra (TRES) and time-resolved area normalized

emission spectra (TRANES) were analyzed to confirm the origin of excitation wavelength dependent

fluorescence of C-dots.[2]

0 20 40 60 80 100

0.000

0.002

0.004

0.006

0.008

0.010

Ab

so

rban

ce

[DNApm

] (pM)

(b)

0 20 40 60 80 100

0.000

0.002

0.004

0.006

0.008

0.010

Ab

so

rban

ce

[DNAmm

] (pM)

Fig.1 Langmuir plots of the absorbance versus concentration for a normal and single-mismatched DNA.

1. Yi-Ju Yao and King-Chuen Lin, Anal. Chim. Acta, 821, 1(2014) (Featured Article and Cover

Picture)

2. N. Dhenadhayalan, K. C. Lin, R. Suresh, and P. Ramamurthy, J. Phys. Chem. C, in press

(2016).


http://ntuchemkclin.weebly.com/research-interest.html

INV-06 52


(Electro)chemiluminescence of N-(aminobutyl)-N-(ethylisoluminol) and Catalyst

Bifunctionalized Gold Nanoparticles and their Applications in Analytical Chemistry

Hua Cui*, Jiangnan, Shu, Lingfeng Gao, F. Li and Mengxiao Liu

CAS Key Laboratory of Soft Matter Chemistry, Collaborative Innovation Center of Chemistry for Energy

Materials, Department of Chemistry, University of Science and Technology of China, Hefei, Anhui, 230026,

China. E-mail: [email protected]; Fax: +86-551-63600730; Tel: +86-551-63600730

In recent years, attention has been paid to chemiluminescence (CL) functionalized gold nanoparticles

(CL-GNPs). CL reagents, such as luminol and N-(aminobutyl)-N-(ethylisoluminol) (ABEI), have been

successfully used for the preparation of CL functionalized gold nanoparticles (GNPs). They have shown

application potentials in the fields such as clinical diagnosis, food safety, environmental monitoring.

More recently, we proposed a new concept of grafting catalyst metal ions on the surface of CL-GNPs by

simply stirring metal complexes with CL-GNPs.1 Herein, we report recent progress in our research

group about CL and electrochemiluminescence (ECL) of Co2+ complexes/ABEI-GNPs and their

analytical applications. Co2+ complexes/ABEI-GNPs exhibited excellent CL activity when they reacted

with H2O2 solution. The CL intensity of Co2+ complexes/ABEI-GNPs was over three orders of

magnitude higher than ABEI-GNPs. Moreover, Co2+ complexes/ABEI-GNPs could directly generate

outstanding ECL emission in neutral and alkaline media without additional coreactant.2 The ECL

emission of Co2+ complexes /ABEI-GNPs was over 2 orders of magnitude higher than that of ABEI-

GNPs in neutral and alkaline media. On this basis, label-free methods were developed for the

determination of pyrophosphate ion and specific DNA sequences from mycobacterium tuberculosis,

hepatitis B virus (HBV) and myelocytomatosis viral oncogene (v-myc).3

REFERENCES

1. M.X. Liu, H.l. Zhang, J.N. Shu, X.Y. Liu, F. Li, and H. Cui. Anal. Chem. 2014, 86, 2857−2861

2. J.N. Shu, W. Wang and H. Cui. Chem. Commun., 2015, 51, 11366-11369

3. L.F. Gao, X. He, L. Ju, X.Y. Liu, F. Li, H. Cui, Anal. Bioanal. Chem., 2016, DOI：10.1007/s00216-015-9244-7

INV-07 53


Fluorescent noble metal nanoclusters for analytical application

Erkang WANG

State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese

Academy of Sciences, P. R. China

University of Chinese Academy of Sciences, P. R. China

E-mail: [email protected]

Noble netal nanoclusters have attracted much research interest because of their unique size-

dependent optical, electronic, magnetic, and catalytic properties. Our group has focused on the

fluorescent noble metal nanoclusters such as silver, gold and cupper nanoclusters for analytical

application. we developed the technique for the detection of DNA, Hg2+ and biothiols based on

DNA protected silver nanoclusters. We have reported photoinduced electron transfer (PET) between

DNA/Ag NCs and G-quadruplex/hemin complexes, accompanied by a decrease in the fluorescence

of DNA/Ag NCs. The novel PET system enabled the specific and versatile detection of target

biomolecules such as DNA and ATP with high sensitivity. Recently, we have developed a high-yield

synthesis of silver nanoclusters using DNA monomers as the scaffolds for the first time. In the

study, we have explained the mechanism of the formation of DNA-protected silver nanoclusters and

the reason why cytosine-rich DNA strands are good scaffolds for fluorescent silver nanoclusters by

using density-functional computations. Our results provide basic guidelines for further experimental

and theoretical studies on DNA-protected fluorescent silver nanoclusters and may ultimately

contribute to the programmed synthesis of DNA-stabilized silver nanoclusters with

photoluminescence properties. We found the AIE(Aggregate induced emission)，AIEE(aggregate

induced emission enhancement) phenomena and mechanism with AgNCs as well as CuNCs. We

also developed the ratiometric fluorescence detection of tyrosinase activity and dopamine using

thiolate-protected AuNCs

1. H. Wei, E. K. Wang, Chem. Soc. Rev., 42, 6060-6093 (2013).

2. L. B. Zhang, J. B. Zhu, S. J. Guo, T. Li, J. Li, E. K. Wang, J. Am. Chem. Soc., 135(7), 2403-

2406 (2013).

3. W. W. Guo, J. P. Yuan, Q. Z. Dong, E. K. Wang. J. Am. Chem. Soc., 132(3), 932-934 (2010).

4. X. Yang, L. F. Gan, L. Han, E. K. Wang, J. Wang, Angew. Chem. Int. Ed. 52, 2202-2206(2013).

5. X. Yang, L. F. Gan, L. Han, E. K. Wang, J. Wang, Chem. Sci. 4 4004-4010(2013).

6. Y. Teng, X. F. Jia, J. Li , E. K. Wang, Anal. Chem., 87, 4897-4902 (2015).

ACKNOWLEDGMENT

This work was supported by the National Natural Science Foundation of China

INV-08 54


Electrochemiluminescence (ECL) and Quartz Crystal Microbalance (QCM)

biosensors based on electrochemical grafting of diazonium compounds

Sun-Min Cho, So-Ra Lee, and Seong-Ho Choi*

Department of Chemistry, Hannam University, Daejeon 305-811, Republic of Korea


It is also possible to functionalize carbon surface with diazonium salts forming an aryl monolayer.

We fabricated ECL biosensor for determining acetylcholine in human blood and QCM biosensor

for recognition of chiral compounds by electrochemical grafting of diazonium compounds. The

prepared the ECL and QCM biosensors were characterized about stability, detection limits, sensing

range, and etc. to electrochemical signals and ECL signals. From these results, we will discuss

about the detailed preparation, characterization and determination of target compounds for ECL

biosensor with CdSe QDs and Ru(bpy)32+ complexes and QCM biosensor with beta cyclodextrin.

INV-09 55


Method developments for the determination of cyanide, thiocyanate, and the mixture

of both, cyanide and thiocyanate based on the flow injection technique.

Hermin Sulistyarti

Research Centre for Low Cost and Automated Method and Instrumentation (LCAMIA), University of

Brawijaya, Malang, 65145, Indonesia

Department of Chemistry, Faculty of Science, University of Brawijaya, Malang, 65145, Indonesia


The extremely toxic cyanide has been utilized in large quantities in many industrial applications,

and to date, cyanide remains unchallenged reagent for gold extraction in milling ores. The deadly

poison cyanide is also generated naturally in plants containing cyanogenic glucoside or found to be

used as an agent of killing. Meanwhile, cyanide often presents together with thiocyanate in the real

world samples, because there is a connection reaction between both compounds. Cyanide can be

produced by either reaction of nitrite with organic compounds or oxidation of thiocyanate by

oxidizing agents; on the other hand, cyanide can be converted to thiocyanate by sulphur compounds

or enzymatically in the body by rhodanese enzyme. Thus, it can result to false report for cyanide or

thiocyanate determination. The milestone methods development for determination of cyanide,

thiocyanate, as well as their sequential determination using flow injection technique combined with

gas diffusion and pervaporator devices as well as ligand exchange technique will be discussed

thoroughly to give a better report for cyanide and thiocyanate determination. All of the presented

methods provide a fast analysis, sensitive, and reproducible results and have been validated with

satisfactory accuracy and precision.

INV-10 56


Paper-based analytical devices for titrations in environmental chemistry

and food chemistry

Takashi Kaneta*, Shoko Miki

Department of Chemistry, Graduate School of Science and Technology, Okayama University,

Okayama 700-8530, Japan


Paper-based analytical devices (PADs) have attracted many attentions in the fields of point-of-care

testing and on-site analysis. Several advantages of the PADs are considered to be their lightness,

inexpensiveness, and transportability. Furthermore, the PADs facilitate chemical analysis outside

the laboratory if we need only small expendable items in the experimental operations. In fact, we

have previously developed novel PADs for acid-base titrations and chelate titrations which

permitted on-site analysis [1,2]. These PADs are superior to classical titrations in terms of speed,

transportability, and inexpensiveness. The most attractive performance of the PAD is to obtain

analytical results without any electric devices since the endpoint can be directly determined by the

number of the detection zones which change the color of an indicator, i.e., it is found by naked eyes

as well as the classical titrations.

These PADs were employed for on-site analysis in environmental chemistry as being demonstrated

by the determination of the acid concentration in natural hot spring water [1] and simultaneous

determination of magnesium and calcium ions in drink water, river water, and seawater [2]. So, the

PADs would also be applicable to food chemistry, e.g., the measurement of the content of citric acid

(or other organic acids) in citrus fruits. Therefore, we attempted to employ the PADs for acid-base

titrations in the measurement of citric acid in this study.

When using the PADs for the measurement of citric acid, we encountered a problem in the

reproducibility of the analytical results. Citric acid is the trivalent acid with pKa values of 3.14,

4.75, and 6.40. When sodium hydroxide was used as a base, the pH at the equivalent point was

estimated to be ~9 which is near the edge of the indicator range for phenolphthalein. Although

phenolphthalein is generally employed for detecting the endpoint in the classical titration of citric

acid, the color change is unclear in the PAD due to a small pH jump around the endpoint. For

example, when we measured a 50 mM citric acid solution using the PAD which was prepared for

the determination of citric acid in the concentration range of 35 to 80 mM at 5 mM interval, the

endpoints were fluctuated between 45 to 55 mM in multiple measurements.

To solve this problem, we tried a back titration using the PAD with an improved channel design.

The PAD has ten channels with three zones which were radially located: the first zones contained a

sufficient amount of sodium hydroxide to neutralize citric acid in the sample, the second zone

contained different amounts of sulfamic acid, and the third zones contained a constant amount of

phenolphthalein. When citric acid was introduced into the center of the PAD, citric acid was

neutralized in the first zones by sodium hydroxide. The residual hydroxide ions penetrated into the

second zones where excess amounts of hydroxide ions were neutralized by sulfamic acid. Finally,

the residual hydroxide ions or hydrogen ions enter into the third zones, resulting in the color change

at the zones where hydroxide ions remained. We optimized the conditions including the

concentrations and volumes of sodium hydroxide, sulfamic acid, and phenolphthalein to improve

the reproducibility of the PADs for neutralization titration of citric acid.

References

[1] S. Karita, T. Kaneta, Anal. Chem., 86, 12108–12114 (2014).

[2] S. Karita, T. Kaneta, Anal. Chim. Acta., 924, 60–67 (2016).

Keywords Paper-based analytical device; On-site analysis; Citric acid; Back titration

INV-11 57


Assessment of co-benefits of emission reduction measures in Southeast Asia on air

quality improvement and climate forcing mitigation

Didin Agustian Permadi1, Nguyen Thi Kim Oanh1* and Robert Vautard2

1 Environmental Engineering and Management, School of Environment, Resources and Development, Asian

Institute of Technology, Klong Luang, Pathumthani 12120, Thailand. 2 Laboratoire des Sciences du Climate de l’Environment (LSCE), Institut Pierre Simon Laplace (IPSL),

Gif Sur Yvette, France.


Updated emission inventory of BC and other species for Southeast Asia (SEA) was prepared for

base year 2007 focusing on man-made sources, including open burning, in Indonesia, Thailand and

Cambodia (BY2007). Two emission scenarios for 2030 were developed to represent business as

usual (BAU2030) and emission reduction measures in four (4) major anthropogenic sources

(RED2030) implemented in Indonesia and Thailand. WRF/CHIMERE modeling system was used

to simulate PM10, PM2.5 and black carbon (BC) air quality over SEA region for the base year of

2007. BAU2030 and RED2030 emission scenarios were simulated using future boundary conditions

taken from global chemistry transport model of LMDZ/INCA to quantify the impacts and co-

benefits on human health and BC climate forcing. Extended aerosol optical depth module

(AODEM) was used to calculate the total columnar aerosol optical depth (AOD) and BC AOD with

the internal mixing assumption. The modeling results were evaluated using available ground-based

observations of aerosol concentrations and AOD from both ground based (AERONET) and

Moderate Resolution Imaging Spectroradiometer (MODIS) satellite product. Even though simulated

ground-level aerosol concentrations were significantly underestimated but model still successfully

fulfilled the suggested criteria for calculated statistical measures especially for PM10 and BC.

PM10 and PM2.5 spatial distribution showed strong seasonal influence of biomass open burning

over Sumatera and Borneo Island during the period of August-November and over Central and

Northern part of Thailand during the period of January-April. Spatial distribution of BC showed

high concentration in SEA big cities that imply significant influence of anthropogenic emission of

fossil fuel combustion. BAU2030 simulation revealed potential increase of premature death in the

domain of 30/100,000 population while RED2030 would potentially avoid total premature death of

59/100,000 population. Under the BAU2030, maximum annual average BC radiative forcing would

increase by 1 W/m2 from 2007 but RED2030 would help to bring the level down by about 0.6

W/m2, hence co-benefits. Future works should focus on regular update of emission inventory and

impacts of future ozone on crops should be included in the co-benefit analysis.

Keywords: WRF/CHIMERE, co-benefits, black carbon, radiative forcing, premature death

INV-12 58


Portable analytical platforms: Instrumental and non-instrumental approaches

Mirek Macka

School of Physical Sciences and Australian Centre for Research on Separation Science (ACROSS),

University of Tasmania, Hobart, TAS, Australia

The demand for and impact of portable analytical technologies is set to grow driven by several

factors including need for rapid on-site answers, and new trends such as wearable devices.

Miniaturisation and advances in technology have opened up analytical techniques traditionally seen

as lab bench bound, such as separation methods, to portability and thus on-site use. At the same

time new concepts have been developed allowing low-cost analytical approaches, such as based on

paperfluidics. The approaches to portable analysis can be very different in the level of

instrumentation involved.

In this presentation the growing applicability of analytical methods to portable on-site analysis will

be illustrated using examples from the literature and from own research, including but not limited to

new solid state light sources such as sub-250 nm LED [1], miniaturised capillary liquid

chromatography (LC) [2], and novel approaches in the area of paperfluidics based analytical

devices [3].

While ideally ‘non-instrumental’ analysis does not exist, the involvement of various type,

sophistication of use, costs etc. of instrumentation will form substantial practical differences

between the various approaches. The speaker will endeavour to interrogate some of the factors

influencing the use of analytical methods in a portable format, and relate them to their use in

various areas. Despite the fact that such discussion is almost certainly set to produce a variety of

opinions and more questions than clear answers, it may produce some insights in future advances in

portable analysis.

[1] Y. Li, P. Nesterenko, B. Paull, R. Stanley, M. Macka. Performance of a new 235 nm UV-LED

based on-capillary photometric detector. Anal. Chem. in print (2016)

[2] Y. Li, M. Dvořák, P. Nesterenko, R. Stanley, N. Nuchtachvorn, L. Kujovská Krčmová, J.

Aufarová, M. Macka. Miniaturised medium pressure capillary liquid chromatography system with

flexible open platform design using off-the-shelf microfluidic components. Anal. Chim. Acta 896

(2015) 166-176.

[3] N. Nuchtachvorn, M. Macka. A novel, highly flexible, simple, rapid and low-cost fabrication

tool for paper- -house

formulated aqueous inks. Anal. Chim. Acta 919 (2016) 70-77.

INV-13 59


Dopant type and amount governs the electrochemical performance of graphene

platforms for the antioxidant activity quantification

Kai Hwee Hui, Adriano Ambrosi, Zdeněk Sofer, Martin Pumera, Alessandra Bonanni*

Division of Chemistry & Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang

Technological University, Singapore 637371

Graphene doped with heteroatoms can show new or improved properties as compared to the

original undoped material. It has been reported that the type of heteroatom and the doping

conditions can have strong influence on the electronic and electrochemical properties of the

resulting material. Here we wish to compare the electrochemical behavior of two n-type and two p-

type doped graphenes, namely boron-doped graphenes and nitrogen-doped graphenes containing

different amounts of heteroatom. We show that the boron-doped graphene containing the higher

amount of dopant provides the best electroanalytical performance in terms of calibration sensitivity,

selectivity and linearity of response for the detection of gallic acid normally used as standard probe

for the quantification of antioxidant activity of food and beverages. Our findings demonstrate that

the type and amount of heteroatom used for the doping have a profound influence on the

electrochemical detection of gallic acid rather than the structural properties of the material such as

amounts of defects, oxygen functionalities and surface area. This finding has a profound influence

on the application of doped graphenes in analytical chemistry field.

INV-14 60


Simple Identification of Geographical Origins of Agricultural Products

using M-13 Bacteriophage Based Structural Color Sensor

Daun Seol1, Youngjong Kang1,Kyungjoon Cha2, Jin-Woo Oh3, Hoeil Chung1*

1Department of Chemistry, College of Natural Sciences, Hanyang University, Seoul, Korea

2Department of Mathematics, College of Natural Sciences, Hanyang University, Seoul, Korea 3Department of Nanoenergy Engineering, Pusan National Universit1, Korea


Identification of geographical origins of agricultural products is an important issue in many

countries for fair evaluation of their commercial values. Since large numbers of samples are

necessary to be analyzed in a practical field, a simple and fast analytical method with capability of

at-site measurement is critically demanding. Although spectroscopic methods such as near-infrared

(NIR) spectroscopy have been frequently adopted for this purpose, it still requires fairly expensive

instrumentation as well as effort of maintenance. Here, we have demonstrated a simple and

versatile color sensor based on M-13 bacteriophage as an alternative analytical tool. M-13

bacteriophage is a filamentous bacteriophage that is mainly composed of 2700 copies of helically

arranged pVIII major coat protein. Typical M-13 bacteriophage is highly regular with a length

of∼880 nm and a diameter of ∼6.6 nm. The sensor was fabricated using a simple pulling technique

by self-assembly of M-13 bacteriophage. When the sensor is exposed to vaporized molecules, M-13

bacteriophage bundles promptly swell and promote distinct structural color change. As generally

known, compositions of agricultural products vary depends on cultivation of area; therefore, the

degree of color change would be different depends on geographical origins of samples. Using the

prepared sensor, we have attempted to differentiate the geographical origins of 5 different

agricultural samples (garlic, onion, red pepper powder, perilla and balloon flower root). Two groups

of domestic and imported samples were included in each case. Vapor from a sample was simply

allowed to contact with the sensor in a chamber and the color change was continuous recorded with

the increase of chamber temperature. Then, principal component analysis was performed using the

obtained signal from each sample and the subsequent scores were finally used for the discrimination

based linear discriminant analysis (LDA). The resulting discrimination accuracies were evaluated

and the advantage/disadvantage of the sensor was discussed in comparison with other analytical

methods.

Keywords: M-13 BACTERIOPHAGE; STRUCTURAL COLOR SENSOR; GEOGRAPHICAL ORIGINS

INV-15 61


Towards whole-animal drug screen targeting cardiovascular diseases

- Spectro-imaging assessment of zebrafish

Ian Liau

Department of Applied Chemistry and Department of Biological Science and Technology, National Chiao

Tung University, Hsinchu 30010, Taiwan

Using animal models is an inevitable component of pharmaceutical development and translational

research. The zebrafish (Danio rerio) has recently emerged as a popular model organism because of

its numerous attractive features including convenience of drug delivery, rapid development, ease of

genetic manipulation, low cost of maintenance and high degree of genetic and functional

conservation of zebrafish relative to human beings. The translucency of the body of larvae further

makes spectro-imaging interrogation of the pathophysiological change of the cardiovascular system

become possible. In this presentation, I will report our recent effort on the development of spectro-

imaging methods and zebrafish disease models towards whole-animal drug screen targeting

cardiovascular diseases. Acute thromboembolic diseases remain the major global cause of death or

disability but therapeutic options remain limited. To this end, we developed a novel zebrafish model

of focal ischemic stroke using photochemical thrombosis; such model can prospectively facilitate

the screen of thrombolytic drugs [1]. Direct interrogation of the vascular lesions in vivo is important

in the assessment of novel antiatherosclerotic therapies. With image-guided Raman spectroscopy,

we delicately probed individual vascular fatty lesions on the zebrafish model of hyperlipidemia, and

demonstrated the pleotropic therapeutic (suppressing the accumulation and oxidation, and

expediting the clearance, of vascular lipids) efficacies of statin, which is notably consistent with the

known pharmaceutical effects of this drug on human beings [2]. Despite the importance in advaning

the use of zebrafish for drug screen, functional assessment of the zebrafish heart remains

challenging because of the small size (~200 μm) and high beating rate (~200 beats per min) of the

larval zebrafish heart. We developed all-optical methods (pseudodynamic 3D imaging and dual-

beam reflectometry) to determine the cardiac function and rhythm of zebrafish, and demonstrated

potential applications in the functional screening of cardioactive and cardiotoxic drugs [3]. Through

collaboration, we synthesized a ‘fluorescent dopamine’ that possesses essential features of natural

dopamine (selective labeling of dopaminergic neurons in vitro and in living zebrafish and being

evoked to release) [4], and developed an optically elicitable polymeric nano-carrier that can be

triggered to release nitric oxide (NO) and was shown to induce vasodilation in the cerebral blood

vessels of zebrafish; such a strategy should help reveal the complex biological effects of NO in vivo

and open up new clinical applications of NO [5]. Taken these results together, we conclude that our

cross-disciplinary approach will not only improve our understanding of the pathophysiology of

human cardiovascular diseases but also facilitate the development of therapeutic interventions

targeting these critical diseases.

References:

1. I.-J. Lee et al. J. Biophotonics 2016 (in press).

2. C.-Y. Yang et al. Anal. Chem. 86, 3863, 2014.

3. K.-Y. Lin et al. Anal. Chem. 86, 2213, 2014; Y.-C. Lai. et al. Biomed. Opt. Exp. 5, 1616, 2014.

4. H.-J. Lin et al. Chem. Comm. 51, 14080, 2015; H.-J. Lin et al. RSC Adv. 6, 71589, 2016.

5. P.-T. Kao et al. Chem. Sci. 2016 (in press).

INV-16 62


In Vivo Detection of Damaged Bone

Lehui LU，Ying WANG, Chunhuan JIANG


Academy of Sciences, 5625 Renmin Street, 130022 Changchun, China

Achieving high-resolution imaging of bone-cracks and even monitoring them in live organisms are

of great significance for understanding their extreme biological effects but remain quite

challenging, especially only adopting commercial imaging systems. We explore the use of clinical

gemstone spectral computed tomography (GSCT) technique as a powerful tool for targeted imaging

of bone-cracks in rats via intramuscularly administrating crack-targeted ytterbium-based contrast

agents (CAs). The superior imaging capability of GSCT allows us to real-time monitor the targeting

and accumulation of CAs towards bone-crack in vivo. Moreover, highly sensitive and non-invasive

in vivo detection of bone crack can be achieved by surface-enhanced Raman scattering (SERS)

technique.

Acknowledgement. Financial support by National Natural Science Foundation of China

(21635007) and National Key Research and Development Program of China (2016YFA0203200) is

gratefully acknowledged.

References

[1] Ying Wang, Chunhuan Jiang, Wenhui He, Kelong Ai, Xiaoyan Ren, Lin Liu, Mengchao

Zhang*, Lehui Lu*, ACS Nano, 2016, 10, 4164-4172.

[2]Yanlan Liu, Kelong Ai, Lehui Lu*, Acc. Chem. Res., 2012, 45, 1817-1827.

[3]Yanlan Liu, Kelong Ai, Lehui Lu*, Chem. Rev., 2014, 114, 5057-5115.

INV-17 63


Urinary 1-hydroxypyrene as a biomarker of exposure to polycyclic aromatic

hydrocarbons: a study of kindergarten children in Chiang Mai Province, Thailand

Tippawan Prapamontol1*, Tanyaporn Kerdnoi1, Surat Hongsibsong1, Ubonwan Chaimongkol1, Sawaeng

Kawichai1, Warangkana Naksen1, Kunrunya Sutan1, Suchart Kiatwattanacharoen2

1Environment and Health Research Unit, Research Institute for Health Sciences, Chiang Mai University,

Chiang Mai, 50200, Thailand 2Department of Radiologic Technology, Faculty of Associated Medical Sciences, Chiang Mai University,

Chiang Mai 50200, Thailand


Outdoor air pollution has been classified by the International Agency for Research on Cancer

(IARC) as carcinogenic to humans (Group 1). Since 2007, upper northern Thai population have

exposed to airborne particulates during smoke haze episode on top of other environmental pollution

background. Airborne particlates are an efficient vehicle of toxic chemicals including polycyclic

aromatic hydrocarbons (PAHs) as piggybacking into human body via inhalation absorption.

Kindergarten children are among the most vulnerable groups to expose to environmental hazards so

we explored the exposure to PAHs by measuring 1-hydroxypyrene (1-OHP) in their urine samples.

The study design was a cross-sectional study of 3 distinct characteristics of kindergarten children

from an urban, a periurban and a rural schools. The total subjects of 160 children were enrolled

from an urban school in Chiang Mai City (urban school children, n=51), 2 schools in periurban and

rural districts of Chiang mai Province (periurban school children, n=50, rual school children, n=59).

The study was conducted during the smoke haze episode from 28 February to 3 Matrch 2011.

Urine, a non-invasive specimen, samples were collected from individual children during school

hours (8.30 – 15 hrs) for 2 days, one on Monday or Tuesday representing weekend exposure and on

Wednesday or Thursday representing weekday exposure. Two-day urine samples were composited

as a sample of individuals. Aliqouts, 10 mL, of composited urines were kept at -20 C in the freezer

until analysis. The method for analysis of urinary 1-OHP has been developed and validated prior

sample analysis using high performance liquid chromatography- fluorescence detection (HP1100,

USA). Proficiency testing of urinary 1-OHP analysis was performed by participation with the

German Extermal Quality Assessment (G-EQUAS). Urinary creatitine is analyzed by the Central

Laboratory of Maharaj Nakorn Chiang Mai Hospital, Chiang Mai University and used to formalize

the detected 1-OHP concentrations. School girls were slightly more than boys as from urban,

periurban, and rural schools were 59, 52, and 56%, respectively. The most common age group was

6 years old with 72.5, 84, and 39%, respectively. Mean±SD urinary 1-OHP concentrations in urban,

periurban, and rural children were 95.8±92.7, 158.7±88.8, and 1,240±733.8 nanomole/mole

creatitine, respectively. They were significantly different (p<0.05) and greater than the Natioanl

Health and Nutrition Examination Survey (NHANES, USA) for 1.5, 2.7, and 20 fold, respectively.

The present study results showed that kindergarten children in the rural area might have exposed to

PAHs and warrents further investigation of exposure sources in order to safeguard their health in

the future.

Keywords 1-hydroxypyrene; biomarker; polycyclic aromatic hydrocarbons; air pollution;

kindergarten children


INV-18 64


Development of monitoring method of Maillard reaction product in milk

Hiroyuki Ukeda1*, Tomoko Shimamura2*, Vu Thu Trang3

1Center of Regional Collaboration (CRC), Kochi University, Kochi, Japan

2Faculty of Agriculture and Marine Science, Kochi University, Kochi, Japan 3Hanoi University of Science and Technology, Hanoi, Vietnam

*E-mail: [email protected]. [email protected]

The Maillard reaction (nonenzymatic glycation) is a common chemical reaction which occurs in

various foods (containing milk and dairy products) during food processing and storage, and

generates multiple reaction products. In the case of milk, lactose mainly reacts with free amino acid

in side chains of milk proteins (such as casein, α-lactalbumin, and β-lactoglobulin) by the

Maillard reaction. Consequently, the Maillard reaction can affect the function of milk proteins and

sensory property of milk and dairy products. Therefore, the detection of the Maillard reaction

products plays an important role in the quality management.

We proposed a novel microplate assay for determining the ability of milk to reduce tetrazolium salt

XTT as a method of evaluating the extent of Maillard reaction1). The XTT assay could estimate not

only heat-treatment condition of milk but also storage condition after heat-treatment. In addition,

we demonstrated the generation and presence of aminoreductone, one of the early-stage Maillard

reaction products, in milk using the XTT assay. In this presentation, detailed process of

development of XTT assay and its application to dairy industry will be introduced. Also we indicate

that the aminoreductone detected by the XTT assay has some functions such as antioxidant activity,

antimicrobial activity, and inhibiting activity of riboflavin photo-degradtion.

1) T. Shimamura & H. Ukeda (2012). Maillard reaction in milk -effect of heat treatment-. In W. L.

Hurley (Ed.), Milk protein (pp. 147-158). Rijeka, Croatia: InTech.

Keywords Maillard reaction, milk, dairy product, aminoreductone, food function

INV-19 65


Surfactant micelles as stationary phases for separation and enrichment

Joselito P. Quirino*

Australian Centre for Research on Separation Science, School of Physical Sciences−Chemistry, University of

Tasmania, Tasmania 7001, Australia


The use of surfactant micelles for analytical separations such as in micellar liquid chromatography

and micellar electrokinetic chromatography are quite well understood. Micelles have also been

widely utilised for analyte enrichment prior to quantitative chemical analysis. Separation and

enrichment relies on the power of surfactant aggregates or micelles to solubilise various

compounds. The use of micelles in separations and enrichments is one of the driving forces that

facilitates the translation of green chemistry to analytical chemistry. Here, new approaches for

analyte separation and enrichment using surfactant micelles in solution and at interfaces will be

presented.

Keywords Surfactant micelles; Liquid chromatography; Micellar electrokinetic chromatography

http://pubs.acs.org/author/Quirino%2C+Joselito+P


INV-20 66


High throughput titration by air-segmented flow ratiometry

Hideji Tanaka1*, Erina Tomiyama1, Tomoko Hirasaka1, Hiroya Kubo1, Sawako Oka1, Naoya Kakiuchi1,

Akihiro Fujikawa2, Masaki Takeuchi1

1Faculty of Pharmaceutical Sciences, Tokushima University, Tokushima, 770-8505, Japan

2Shikoku Riken Co., Tokushima, 771-0124, Japan


Feedback-based flow ratiometry, originated by Tanaka and Dasgupta [1], is a new paradigm of on-

line true titration. The effect of the lag time (tlag) between the titrand-titrant confluence and the

detection downstream is compensated for by averaging rapid forward and backward titrations. The

throughput rate is ca. 20 titrations/min. Tanaka et al. [2,3] further developed this approach by

coupling the feedback control with subsequent fixed triangular wave control. The throughput rate is

as high as 34 titrations/min [3]. In the present study, air-segmentation and software-based phase

recognition [4] are introduced to the feedback-based - fixed triangular wave controlled flow

ratiometry. The aim of the air introduction is to acquire signals that accurately reflect the flow

ratios at respective minute domains of the stream. An in house program written in Excel VBA was

used to acquire, analyze and display the data automatically. In the measurement, the flow rate of a

titrant (eg. NaOH) containing an indicator (eg. Bromothymol Blue) is linearly varied with a control

voltage (Vc) generated from a computer. The titrant is merged with a titrand (eg. HCl) while the

total flow rate is held constant. Air is introduced to the merged liquid stream in order to limit the

dispersion within each liquid segment separated by air bubbles. The stream is led to a detector with

no physical deaeration. The detector output signals (Vd; absorbance or transmittance) are acquired

in a computer as a Microsoft Excel format. Air signals are removed by a signal processing.

Initially, a feedback-based control is applied, where the scan direction of Vc is reversed from

upward to downward, and vice versa, whenever the transition of the indicator at the equivalence

point is sensed. Averaging of the most recent maximum Vc and the minimum Vc can offset the

influence of tlag and gives the value of Vc that yields the equivalence composition at the confluence

point. Next, the scan range of Vc is further limited to the range just around the equivalence point by

using fixed triangular wave having higher scan rate so as to increase the throughput rate. The

threshold value for the second derivative of Vd is found to be effective to distinguish sharp air

signals from liquid signals, especially from those around the equivalence point. In this conference,

we will present the advantages and prospect of the air-segmented flow ratiometry.

[1] H. Tanaka, P.K. Dasgupta, J. Huang, Anal. Chem., 72 (2000) 4713.

[2] H. Tanaka, T. Baba, Talanta, 67 (2005) 848.

[3] H. Tanaka, T. Baba, Anal. Sci., 21 (2005) 615.

[4] T. Ogusu, K. Uchimoto, M. Takeuchi, H. Tanaka, Talanta, 118 (2014) 123.

Keywords Flow analysis; Flow ratiometry; Titration; Air-segmentation

INV-21 67


Preparation of organic polymer-based monolithic column for flow-through protein

digestion and phosphopeptide identification

Akhmad Sabarudin1,2

1Department of Chemistry, Faculty of Science, Brawijaya University, Malang, Indonesia 2Research Center for Advanced System and Material Technology, Brawijaya University, Malang, Indonesia

E-mail: [email protected] ; [email protected]

Two kinds of organic polymer-based monolithic microreactors were prepared for proteomic

application. A poly (glycidyl methacrylate-co-ethylene glycol dimethacrylate) (poly(GMA-co-

EDMA)) monolith as a nanoporous polymeric-support was synthesized by in-situ copolymerization

inside the microbore silicosteel tubing (1.0 mm i.d x 100 mm). This polymeric-support was chemically

modified with trypsin through the ring-opening reaction of epoxide group to form a nanobiocatalyst

microreactor (TIMM). Another microreactor was prepared by immobilizing Ti4+ onto the monolith

after post-modification with aminomethyl phosphonic acid (Ti4+-IMAC). The properties of the

monolithic miroreactors, which involve morphology, elemental analysis, surface area analysis,

permeability and pore distribution were characterized in detail. It was found that the optimized

monolith has good mechanical stability and high permeability, accompanied with the ideal proportion

of macropores and mesopores. The optimized TIMM was successfully applied to a rapid and efficient

digestion of protein samples using flow-through method, whereas Ti4+-IMAC showed good selectivity

for collection of phosphopeptides from digested proteins and purification of tyrosine phosphorylated

peptide samples.

Keywords: Monolith, Immobilized Enzyme, protein, phosphopeptide, titanium, flow-through method

INV-22 68


How to stimulate scientific imagination in undergraduate Chemistry class

Hitoshi Watarai

International College, Osaka University, and Institute for NanoScience Design, Osaka University, Japan


Introductory Chemistry in undergraduate course is sometimes hard to organize, especially for

students who were not taking Chemistry in high school. It will be true that some discontinuity

presents between high school chemistry and University chemistry. For example, the image of

electrons has to be changed in the undergraduate Chemistry to understand the concept of quantum

chemistry. Thermodynamic view of the chemical equilibrium is also important but hard to be

understood by non-science major students. I have taught General Chemistry in some Universities,

and also I am teaching Analytical Chemistry in International College of Osaka University (1).

According to my experience, primitive questions are sometimes useful to stimulate their interests in

the material world and refine their knowledge. There are many questions which will be asked about

the properties of electrons and chemical reactions, which are not written in any textbooks.

For example, if one asks about the size of electron, it will cause a little trouble, because there are no

books which describe correctly the diameter of electron, though the diameters of proton, neutron

and hydrogen atom are precisely described. The idea that an electron has no size will cause a new

insight into the image of atoms and molecules. After some discussions, they will be able to imagine

that an atom is almost a “vacuum ball”. Actually, the properties of electron are very strange and

against common sense. Why can an electron in hydrogen atom rotate at the speed of about 1/100 of

velocity of light around proton with no consumption of energy? Is an electron working like a

permanent engine?. Once students accept the idea that the electron has no size and an electron never

stop moving, it seems easier to expect the behavior of electrons in atoms and molecules. This point

should be more emphasized in the general Chemistry class. I am asking such primitive questions to

students very often. Another example of primitive questions is the acid dissociation constant, Ka, of

a single acetic acid molecule in water. We will discuss what will happen in a single acetic acid

molecule in water and how to define the acid dissociation constant for the single molecule. The

value of Ka seems fluctuate between 0 and infinity, since the single acetic acid molecule has to take

the neutral form or the dissociated form. A student may say that it is impossible to think Ka value

for a single molecule. The point is, of course, the probability of dissociation. After discussions,

students will learn the equality of the number averaged equilibrium constant and the time averaged

equilibrium constant. Now, the single molecule detection has become a popular technique and it

will be possible to show the experimental result of a single molecule reaction observed by a

fluorescence microscopy (2).

1.Chemistry-Biology Combined Major Program, Osaka University, http://www.osaka-

u.ac.jp/en/international/action/global30/ chembio_cmp

2. T. Matsui, S. Tsukahara, H. Watarai, Single-Molecule Lactonization of Octadecylrhodamine B at

a Liquid-Liquid Interface, Langmuir (2012), 28, 15428-15432.

Keywords Scientific imagination; Primitive questions; Undergraduate Chemistry class

http://www.osaka-u.ac.jp/en/international/action/global30/chembio_cmp

http://www.osaka-u.ac.jp/en/international/action/global30/chembio_cmp

INV-23 69


Green Analytical Chemistry at work for you

Conrad H. Bergo

East Stroudsburg University, East Stroudsburg, PA 18301, USA


The latest techniques available make analytical chemistry greener and cleaner than ever. The

drawbacks of many traditional solvents will be discussed. Benign chemicals as solvents will be

proposed. Separation techniques have been greatly improved especially liquid chromatography and

ultra high liquid chromatography. Green alternatives for testing and separations will be presented.

Careful choices of techniques and solvents will provide a cleaner environment and save your

laboratory money.

INV-24 70


Double-stranded DNA-functionalized nanoparticle assembly for sensing

Mizuo Maeda1*

1Bioengineering Laboratory, RIKEN, Wako-shi, Saitama 351-0198, Japan


Single-stranded (ss) DNA-functionalized Au nanoparticles (GNPs) disperse in an aqueous medium

due to electrostatic repulsion between anionic phosphate groups in the ssDNA backbone.

Interestingly, when the complementary ssDNA is added to the dispersion of the ssDNA-GNPs to

form the fully matched double strand (ds) on the surface, the GNPs become unstable and

spontaneously form aggregates in a high ionic strength medium with a non-crosslinking manner.

Exploiting the unique colloidal behavior of the dsDNA-GNPs, we developed a highly reliable SNP

detection method by combining single-base primer extension and salt-induced aggregation of

dsDNA-GNPs, since those having a single-base protrusion at the outermost surface disperse stably,

allowing detection of a single-base difference in length by color changes. When the surface-

anchored probe DNA is designed to be specific to the SNP typing primer, the resulting dsDNA-

GNPs work as a visual indicator of single-base extension. A set of four extension reaction mixtures

is prepared using each of ddNTPs and subsequently subjected to the aggregation assay. Only one

mixture with the complementary ddNTP generates the single-base protrusion and appears red

(dispersed). This method should potentially be used for clinical diagnostics in personalized

medicine.

One of the drawbacks of the method is low detection sensitivity (DNA detection limit = 400 nM by

colorimetric detection and 30 nM by microfluidic device). In order to improve the sensitivity, we

employed dark field microscopy (DFM), which can monitor scattered light from individual single

metal nanostructures. DFM imaging was able to visualize and quantify the complementary DNA-

induced gold nanoparticle aggregation at the single-particle level, with a detection limit of 100 fM

DNA.

Through a combination of the non-crosslinking aggregation of dsDNA-GNPs and molecular

recognition by aptamers or aptazymes, we have also developed bioanalytical systems for

specifically detecting cGMP, cAMP, cCMP, ATP, FMN, and theophylline. We have recently

demonstrated that the stably dispersed dsDNA-GNPs with a (sub)terminal T-T or C-C mismatch

site rapidly aggregated when Hg(II) or Ag(I) was added to the dispersion. This is because the

mismatch-enhanced terminal breathing of dsDNA, and thus the interparticle entropic repulsion,

were drastically suppressed by the metal-mediated base pairing, T-Hg(II)-T or C-Ag(I)-C, at the

dsDNA terminal. The phenomenon was successfully applied to heavy-metal ions sensing.

We have prepared the hierarchically organized DNA-GNPs assembly that can be responding

selectively to terminal base-pairing. GNPs (5 nm in diameter) covalently functionalized with one

strand of 35-nucleotide (nt) ssDNA and five strands of 16-nt ssDNA were mixed with a ssDNA

template having three binding sites to afford discrete ssDNA–GNP trimer. We found that the

addition of complementary 16-nt ssDNA with 10 mM MgCl2 resulted in significant decrease of the

interparticle distance in dsDNA-GNP trimers. Interestingly, dsDNA-GNP trimers with the terminal-

base-substituted ssDNA (terminal mismatch) did not cause any changes under identical conditions.

The present approach will allow for creation of stimuli-responsive materials with controlled

morphological features.

Keywords DNA; gold nanoparticle; nanoparticle sensor

INV-25 71


Artificial enzyme mimics based on nanomaterials

Shaojun DONG


Academy of Sciences, Changchun, 130022, China

Artificial enzyme mimics have attracted much attention due to their viable merits over natural

enzymes, such as high stability, easy preparation and low-cost. Since the exciting discovery of

ferromagnetic nanoparticles, various nanomaterial-based artificial enzymes (nanoenzymes) have

been developed during the past few decades.

We firstly found, TiO2 nanotube array (TiO2 NTA) possessed intrinsic peroxidase-like activity.

Kinetic study indicated that the catalysis was in accordance with the typical Michaelis–Menten

kinetics and followed a ping-pong mechanism. In addition, the TiO2 NTA electrode showed

excellent electrocatalytic activity towards H2O2 reduction with a LOD of 50 nM.

A fully integrated bioactive paper based on graphene oxide@SiO2@CeO2 was fabricated, which

can be used to detect glucose, lactate, uric acid and cholesterol simultaneously.

Recently, Ni-Pd hollow nanoparticals showing triple-enzyme mimetic activity ( oxidase-like

activity, peroxidase like activity and catalase-like activity) were found. Accordingly, a simple

glucose biosensor with wide linear range and low detection limit was proposed.

Acknowledgment

This work was supported by the National Natural Science Foundation of China (Nos. 21375123 and

21675151) and the Ministry of Science and Technology of China (Nos. 2013YQ170585 and

2016YFA0203201).

INV-26 72


Prospectives of using diamond based adsorbents in liquid chromatography

Pavel N. Nesterenko

Australian Centre for Research on Separation Science, School of Physical Sciences

University of Tasmania, Private Bag 75, Sandy Bay, Hobart, TAS 7001, Australia


Diamond based materials have become widely available following advances in their preparation.

The combination of a unique range of properties, such as chemical inertness, mechanical, thermal

and hydrolytic stability, makes diamond a promising material for use in various modes of liquid

chromatography [1,2]. At the same time, the preparation of efficient chromatographic phases for

liquid chromatography is not trivial task, especially, in terms of obtaining materials having a

developed porous structure and uniform surface chemistry. The presentation provides

comprehensive overview of various approaches for the preparation of diamond based adsorbents

with focus on their chromatographic performance adsorption properties and possible applications in

various modes of high-performance liquid chromatography (HPLC).

The maximum achieved separation efficiency for the column packed with diamond particles is

about 110,000 theoretical plates per meter obtained in ultra-high pressure liquid chromatography

(UHPLC) mode. The possibility of high-temperature liquid chromatography (HTLC) at column

temperatures up to 200oC was also demonstrated. The unique selectivity of diamond was observed

in normal-phase HPLC and HILIC modes.

This research was supported under Australian Research Council's Discovery Projects funding

scheme (Discovery Grants DP110102046 and 150102608).

[1] P.N. Nesterenko, P.R. Haddad. Diamond-related materials as potential new separation media in

separation science. , Anal. Bioanal. Chem. 396 (2010) 205-211.

[2] A. Peristyy, O.N. Fedyanina, B. Paull, P.N. Nesterenko. Diamond based adsorbents and their

application in chromatography. J. Chromatogr. A. 1357 (2014) 68-86.


INV-27 73


Communication in Analytical Sciences in Asia

Takeaki Ozawa

Editor of Analytical Sciences,

Department of Chemistry, School of Science, the University of Tokyo, Japan

“Analytical Sciences” is an international journal published monthly by The Japan Society for

Analytical Chemistry. The journal publishes papers on all aspects of the theory and practice of

analytical sciences, including fundamental and applied, inorganic and organic, wet chemical and

instrumental methods. We received more than 600 original papers in 2015, of which over 220

papers have been published. An important characteristic of the journal of Analytical Sciences is to

receive original papers mostly submitted from Asian countries. To further improve visibility of this

journal in Asia, we are now trying to strongly reinforce journal contents, group managements and

international cooperation. I herein show these reinforcements with metrics and discuss some

advantages on submitting your original researches in the field of analytical chemistry to the

Analytical Sciences.

INV-28 74


Application of ultrasonic radiation and vortex agitation in

surfactant-based microextraction techniques

Rodjana Burakham

Materials Chemistry Research Center, Department of Chemistry and Center of Excellence for Innovation in

Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand


Sample preparation is generally necessary for determination of analytes in real samples. The

primary objective is to clean-up and/or concentrate the analytes of interest, thus reducing or even

eliminating the potential interferences from sample matrices. Recently, there is an increasing

interest in the development of environmentally friendly sample preparation techniques according to

the approach of green analytical chemistry. Thus miniaturization and improvement of sample

handling using alternatives is a challenge that has been recently developed. The use of surfactants in

low consumption organic solvent-based microextraction techniques is an important contribution to

this strategy. In addition, ultrasonic irradiation and vortex agitation have been introduced to

enhance the efficiency of surfactant-based microextraction techniques. A variety of new

modifications, both solvent-based and sorbent-based microextraction methodologies have appeared.

The presentation will describe the development of microextraction procedures based on using

surfactants as extraction media in combination of applying ultrasonic irradiation or vortex agitation

to accelerate the extraction efficiency. These include ultrasound-assisted surfactant-enhanced

emulsification microextraction (UASEME), vortex-assisted surfactant-enhanced emulsification

microextraction with solidification of floating organic droplet (VASEME-SFO), and vortex-assisted

dispersive micro-solid phase extraction (VA-D--SPE). Design of experimental process and effect

of experimental conditions of the proposed procedures will be discussed. Application of each

developed system for determination of various target analytes in real sample matrices will be

demonstrated.

Acknowledgements: The Thailand Research Fund and Khon Kaen University, the Royal Golden

Jubilee Ph.D. Program, PERCH-CIC and the Materials Chemistry Research Center, KKU are

gratefully acknowledged.

Keywords Sample preparation; Microextraction; Surfactant; Ultrasound; Vortex

INV-29 75


Chemometrics in food analysis: A Malaysian perspective

Sharifuddin M Zain

Chemistry Department, University of Malaya, Malaysia

The question of food quality has always been a priority for consumers nowadays. The physical and

nutritional characteristics, including the authenticity and traceability of food products have to be of

an acceptable level to these consumers. Hence, it is of utmost importance that the authenticity and

traceability of food are in accordance to the labels and descriptions of the product which

consequently are in line with the law regarding food products. The knowledge related to these

quality parameters could be considered a further guarantee of the quality and safety the food

product. In this presentation, the synergistic use of analytical techniques and chemometrics carried

out by selected Malaysian researchers on food quality will be discussed. These methods represent a

promising way to obtain trustworthy results in the development of authenticity and traceability

models. The various methods of chemometrics in resolving some real issues related to food

traceability and authenticity will be thoroughly discussed. Particular attention will be paid to the use

of some exploratory, classification, discrimination and modeling techniques. In the initial part of the

presentation, regulatory issues and the overall scope of the use of chemometrics in the Malaysian

food research will be examined. This will involve some selected research work that had been

carried out in tackling some issues related to food authenticity and traceability. Discussion on the

chemometric methods employed and how these methods are useful in providing the needed quality

related information will be deliberated in the second part of the presentation. Finally opportunities

in proliferating the use of chemometric methods in food analysis in Malaysia will be discussed,

forming the conclusion to the presentation.

INV-30 76


Array-based sensor for multiplex detection

Nitsara Karoonuthaisiri*1, Ratthaphol Charlermroj1, Oraprapai Gajanandana1, Manlika Makornwattana1,

Sudtida Phuengwas1, Orawan Himananta1, Channarong Seepiban1, Thanasat Sooksimuang2

1National Center for Genetic Engineering and Biotechnology,

2National Metal and Materials Technology Center,

National Science and Technology Development Agency, Pathum Thani 12120, Thailand


In the current world where more and more analytes are required to be assessed for quality and

safety assurance of products, array technology becomes one of the most popular platforms to meet

such demand of high-throughout detection. This talk will cover three array-based sensor platforms

(surface array, bead array and lateral flow array) and their wide range of applications emphasized

on food and agriculture.

Surface array was developed by fabricating different specific antibodies at predesignated positions

on a well of a conventional 96-well plate to allow simultaneous detection. To each well, a test

sample was added to allow the contaminants to specifically bind to the fabricated antibodies. A

mixture of fluorescently labeled detecting antibodies was subsequently added and the signals of

each antibody spots were acquired. An application for detection of multiple plant pathogens namely

a fruit blotch bacterium Acidovorax citrulli, Potyvirus, Watermelon silver mottle virus, and Melon

yellow spot virus was successfully demonstrated.

Bead array was developed to enable simultaneous detection of multiple analytes using fluorescently

barcoded magnetic beads. The beads were linked with specific antibodies which were used to

capture corresponding pathogens. The presence of pathogens was detected by R-phycoerythrin

labeled antibodies specific to the analytes. This platform was successfully developed for the four

plant pathogens with substantially higher sensitivity than enzyme-linked immunosorbent assay

(ELISA). The developed assay was validated with field cucurbit samples (cucumber, cantaloupe,

melon, and watermelon), resulting in 100% agreement in term of relative accuracy, relative

specificity and relative sensitivity when compared to the commercial ELISA kits for all pathogens.

Lateral flow array integrated the multiplex capacity of array technology with a novel luminescent

organic fluorescent molecule which could be detected using an inexpensive UV light. Its application

for mycotoxin detection will be presented. This platform moved array-based sensor closer to point-

of-care concept.

The advantages and disadvantages of these platforms will be discussed in details.

Keywords Surface array; bead array; lateral flow array; multiplex detection; plant pathogens


INV-31 77


Design and analytical application of optical bio-logical gate

Xiurong YANG

State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry,

Chinese Academy of Sciences

A molecular Boolean logic gate, conceptually derived from electronic logic gates of conventional

computer microprocessors, is an idea of high interest that inspires the generation of smart chemical

and biochemical systems for diagnostic and biosensing applications. Boolean logic provides a

simple and concise way to describe the output of processes that depend on more than one factor.

Bio-logic gates with the advent of more sophisticated biological tools, both nucleic acid- and

protein-based logic systems have been generated. Among them optical signal outputs can be

visually distinguished or easily monitored via UV-vis absorption spectra or fluorescent signal

outputs.

Several bio-logical gates based on optical detection and their analytical applications had been

studied. A NAND logic gate for the logic detection of nitrite using a fluorescent approach with

BSA-protected gold nanocluster probe had been developed. DNA-based ‘‘OR’’ and ‘‘AND’’

colorimetric logic gates had been constructed, which employed aptamers as recognition units and

unmodified AuNPs as optical sensing elements, for the detection of adenosine and cocaine. A

straightforward one-pot strategy to synthesize fluorescent Ag/Au bimetallic nanoclusters had been

developed. The fluorescence of Ag/Au bimetallic nanoclusters was found to be selectively

quenched by Cu2+ ions. The ensemble was constructed as an integrated logic gate of “INHIBIT”

and “OR”. Under the optimal conditions, histidine and cysteine were detected. An europium

luminescence method for“AND” logic gate had also developed. The luminescence intensity was

defined as output, only the presence of both Ag+ and SCN− could make the gate active. The

introduction of S2− was found to be capable of efficiently quenching the luminescence, which

allowed the construction of a fascinating sensor for S2−.

Optical bio-logical gates are simple and rapid methods, avoiding the tedious and time-consuming

synthetic procedures. They will open up a new perspective for various analytical applications.

Acknowledgement

This work was supported by the National Natural Science Foundation of China (No. 21435005)

INV-32 78


Printed graphene sensors for food safety applications

Adisorn Tuantranont

Thai Organic and Printed Electronics Innovation Center (TOPIC)

National Electronics and Computer Technology Center (NECTEC)

Email: [email protected]

Graphene, emerging as a true 2-dimensional material, has received increasing attention due to its

unique physicochemical properties (high surface area, excellent conductivity, high mechanical

strength, and ease of functionalization and synthesis). Printed Electronic also is a new wave of

large-area electronics and flexible electronics manufactured by printing technology. Novel

chemical/biological nanosensors that has higher sensitivity and lower detection limit can be

produced by using printed graphene technology. The talk is about to show how to synthesize

graphene and use as an ink for printing fabrication of electrochemical electrodes in portable food

safety applications such as detection of aflatoxin in feedstock etc.

INV-33 79


Development of New Materials for Electrochemiluminescence

Guobao XU1,*, Wenjing QI1,2, Ling ZHANG1,2, Zhongyuan LIU1,

Shimeles ADDISU KITTE1,2, Jianping LAI1,2

1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese

Academy of Sciences, Changchun, Jilin 130022, P. R. China 2University of the Chinese Academy of Sciences, Chinese Academy of Sciences


Electrochemiluminescence (ECL), or electrogenerated chemiluminescence, is a luminescence

phenomenon resulting from the combination of electrochemistry with chemiluminescence. Similar

to chemiluminescence, it does not need light source and has relatively low background. The use of

electrochemistry techniques allow the facile temperal and spatial tunning, improve reproducibility,

and expand applications. It has been widely in immunoassays, DNA probe assays, aptasensors, the

determination of numerous coreactants, light-emitting devices, and so on. Luminophores,

coreactants, electrocatalysts and quencheres are frequently used in ECL. The properties of these

materials have significant effect on the performance of ECL[1].

In this talk, our recent progress on the development of some high-performance materials will be

presented, such as new coreactants, noble metal nanocrystal electrocatalyst, and efficient quenchers

(e.g. single-walled carbon nanohorn and ozone ). These materials extend ECL applications, improve

ECL sensitivity or selectivity.[1-10]

Acknowledgements

This project was supported by the National Natural Science Foundation of China (No. 21475123

and 21505128), and CAS President’s International Fellowship Initiative (PIFI).

References

[1] Liu Z, Qi W, Xu G (2015) Recent advances in electrochemiluminescence. Chem. Soc. Rev., 45:

715–752.

[2] Lai J, Niu W, Luque R, Xu G (2015) Solvothermal Synthesis of Metal Nanocrystals and Their

Applications. Nano Today, 10: 240–267.

[3] Shimeles Addisu Kitte K, Wang C, Li S, Zholudov Y, Qi L, Li J, Xu G, Electrogenerated

chemiluminescence of tris(2,2'-bipyridine)ruthenium(II) using N-(3-aminopropyl)diethanolamine as

coreactant. Anal. Bioanal. Chem., 2016, 408: 7059-7065.

[4] Zhang L, Niu W, Gao W, Qi L, Lai J, Zhao J, Xu G (2014) Synthesis of Convex Hexoctahedral

Palladium@Gold Core-Shell Nanocrystals with {431} High-Index Facets with Remarkable

Electrochemiluminescence Activities. ACS Nano., 8: 5953–5958.

[5] Liu Z, Zhang W, Qi W, Gao W, Hanif S, Saqib M, Xu G (2015) Label-free signal-on ATP

aptasensor based on the remarkable quenching of tris(2,2'-bipyridine)ruthenium(II)

electrochemiluminescence by single-walled carbon nanohorn. Chem. Commun., 51: 4256–4258.

[6] Qi W, Liu Z, Lai J, Gao W, Liu X, Xu M, Xu G (2014) Detection of ozone based on its striking

inhibition of tris(1,10-phenanthroline)ruthenium(II)/glyoxal electrochemiluminescence. Chem.

Commun., 50: 8164–8166.

[7] Qi W, Wu D, Zhao J, Liu Z, Zhang W, Zhang L, Xu G (2013) Electrochemiluminescence

Resonance Energy Transfer Based on Ru(phen)32+-Doped Silica Nanoparticles and Its Application

in “Turn-on” Detection of Ozone, Anal. Chem., 85: 3207–3212.

[8] Gao W, Liu Z, Qi L, Lai J, Addisu Kitte S, Xu G. (2016) Ultrasensitive Glutathione Detection

Based on Lucigenin Cathodic Electrochemiluminescence in the Presence of MnO2 Nanosheets.

Anal. Chem., 88: 7654–7659.

INV-34 80


Flow field-flow fractionation with mass spectrometry

for subcellular proteins & metalloproteins

Myeong Hee Moon

Dept. of Chemistry, Yonsei University, Seoul, 13722, Korea


Flow field-flow fractionation (FlFFF) is a separation method capable of size-sorting particles and

biological macromolecules like proteins, DNA, cells, and etc in an empty channel space without

packing materials. Analysis of subcellular proteins is of interests in functional proteomics but requires

an accurate and fast separation/isolation of subcellular organelles such as nucleus, lysosomes,

mitochondrion, peroxisomes, Golgi apparatus, etc which are essential components in cells due to their

own specific roles. Organelle separation has widely been conducted by centrifuge based methods,

fluorescence-activated organelle sorting, and affinity purification methods. This presentation will show

recent studies for the subcellular protein characterization and for the metalloprotein analysis with off-

line or on-line hyphenation of asymmetrical flow FFF (AF4) with mass spectrometry (MS). Separation

of subcellular organelles from human embryonic kidney 293 T (HEK293T) cell line was accomplished

with AF4 without any pre-processing steps. It was conducted in steric/hyperlayer mode first and then

few incompletely resolved fractions were re-injected to AF4 for improved separation in normal mode.

Confirmation of organelles were made with SEM, Western blot, and proteomic analysis using LC-

MS/MS. A new approach to analyze metals in metalloproteins will be introduced by directly

hyphenating a miniaturized AF4 system with inductively coupled plasma mass spectrometry (mAF4-

ICP-MS). Selective analysis of metals in biological systems is an emerging research field and is

important to understand the biological processes such as signal transduction and metabolic pathways.

Though metalloproteins are reported to be potential biomarkers of diseases, critical roles of these metal

ions remain unknown. mAF4-ICP-MS was demonstrated not only to separate metalloproteins in blood

plasma directly, but to quantitatively analyze metals and few non-metal elements simultaneously.

Optimizations in the analysis of blood plasma will be discussed together with applications to lung

cancer plasma samples.

Keywords field-flow fractionation; organelles; metalloproteins; nLC-ESI-MS/MS; mAF4-ICP-MS

INV-35 81


Current Status and Prospect of Chinese Journal of Analytical Chemistry

Fan Yang

Editorial Office of Chinese Journal of Analytical Chemistry and State Key Laboratory of Electroanalytical

Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, No. 5625 Renmin Street,

Changchun Jilin 130022, China


Chinese Journal of Analytical Chemistry (Chin. J. Anal. Chem.) is an academic journal of analytical

chemistry established in 1972 sponsored by the Chinese Chemical Society and Changchun Institute

of Applied Chemistry, Chinese Academy of Sciences. Its publishes original scientific research

achievements and review papers as well as short communications in all areas of analytical

chemistry. The journal sets up 6 columns including Research Papers, Research Notes, Research

letter, Experimental Technique and Instrument, Review and Progress or Summary Accounts. The

journal is published monthly in Chinese language with English abstract, and a total of about 300

papers are published each year from over 1000 submitted manuscripts. A detailed abstract,

keywords and the titles of figures and tables are provided in English. It has been indexed by the

Scientific Citation Index (SCI) since1999 and is the first and sole SCI indexed analytical chemical

journal in China, although the impact factor is not high (Between 0.941 in 2011 and 0.554 in 2015).

Since 2006, the journal cooperates with Elsevier Publisher and publishes English version of the

journal, in which 6-8 articles of each issue are translated into English and published on the website

(www.sciencedirect.com) synchronously. Chin. J. Anal. Chem. is a core scientific journal in China.

The contents of the journal have been indexed and cited in more than 30 abstract or index

publications and databases, such as Science Citation Index (SCI, USA), Chemical Abstracts (CA,

USA), Current Bibliography on Science and Technology (Japan) and Pž (Russia) etc. Since 1994,

the impact factors and the cited numbers of the journal were in the first rank in Chinese scientific

journals from Chinese Journal Citation Report (CJCR). The journal has been continuously honoured

as the “100 Outstanding Academic Journals of China” each year in the past nine years. You are

warmly welcome to submit papers in English to the Chinese Journal of Analytical Chemistry.

Keywords Chinese Journal of Analytical Chemistry; Prospect; Development

http://www.sciencedirect.com/

INV-36 82


Practical applications of flow-based systems to steel, urine and water analyses

Norio Teshima*, Hiroya Murakami, Tadao Sakai

Department of Applied Chemistry, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho,

Toyota 470-0392, Japan


A key feature of flow injection analysis (FIA) is that since all conditions are reproduced, dispersion

is very controlled and reproducible. Sequential injection analysis (SIA) is a computer-controlled,

single-line, injection technique that simplifies the manifold and is more robust for unattended

operation [1]. Stopped-in-loop flow analysis (SILFA) is an alternative stopped flow technique

which a chemical reaction proceeds in the coiled loop on a six-way injection valve; such feature is

suitable for handling a relatively slow chemical reaction and is able to avoid a stain on the flow cell

window caused by a colored reaction product [2–4]. These flow-based analyses are promising

techniques used to obtain a highly sensitive and full automated method, provided that a suitable

chemical reaction is introduced into their systems.

In this lecture, we introduce some practical applications of flow-based techniques: a FI

spectrophotometric method for the determination of phosphorus in steel [5], a SI-based solid phase

extraction/ICP-MS method for the determination of vanadium in urine, and a SILFA method for the

catalytic determination of vanadium and iron in water samples [4].

References

[1] G. D. Christian, P. K. Dasgupta, K. A. Schug, Analytical Chemistry, 7th ed. New York: Wiley,

1988.”

[2] N. Teshima, S. Ohno, T. Sakai, Anal. Sci., 23, 1 (2007).

[3] N. Teshima, M. Kuno, M. Ueda, H. Ueda, S. Ohno, T. Sakai, Talanta, 79, 517 (2009).

[4] A. A.-Quezada, K. Ohara, N. Ratanawimarnwong, D. Nacapricha, H. Murakami, N. Teshima,

T. Sakai, Talanta, 144, 844 (2015).

[5] H. Murakami, S. Kamiya, M. Tsuge, M. Kuzuya, K. Morita, T. Sakai, N. Teshima, Bunseki

Kagaku, 65, 387 (2016).

Keywords Flow injection analysis; Sequential injection analysis; Water analysis, Steel analysis;

Urinalysis

INV-37 83


Chemometric analysis of spectroscopic data : applications of nanoparticle synthesis

and clinical diagnosis

Kanet Wongravee,*a Tewarak Parnklang,a Prompong Pienpinijtham,a

Chutiparn Lertvachirapaiboon,a Mika Ishigaki,b Yukihiro Ozaki,b and Sanong Ekgasita

a Sensor Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok

10330, Thailand. E-mail: [email protected]; Fax: +66 2218 7585; Tel: +66 2218 7585 b Department of Chemistry, School of Science and Technology, Kwansei Gakuin University, Sanda,

Hyogo 669-1337, Japan. E-mail: [email protected]; Fax: +81 79 565 9077; Tel: +81 79 565 8349

Chemometrics is well-known as a powerful multivariate analysis to extract significant and

important information from the data with high complexity. Spectroscopy is one of the most

powerful techniques to investigate the changes at the molecular level including the morphological

changes, structural changes, concentration changes and also residue changes. However, the

technique involves an enormous number of variables such as wavelength, wavenumber, chemical

shift (it can be > 1,000). To interpret the data, the multivariate analysis might be required. In the

study, a combination of the spectrophotometric method and chemometric analysis has potential to

reveal the structure evolution mechanism of silver nanoparticles and also to diagnose Esophageal

cancer.

In case of structural evolution of silver nanospheres (AgNSs) to silver nanoprisms (AgNPrs), the

extinction spectra of the evolving nanostructures were analyzed by factor analysis and error

indicator functions. The resolved concentration profiles and pure spectra of the components were

obtained using multivariate curve resolution-alternative least squares (MCR-ALS) analysis. The

evolution profiles show that the AgNSs systematically evolved into plate structures with different

sizes. Larger nanoplates were obtained when higher concentration of H2O2 were employed. The

predicted structural morphologies of each component given by chemometic calculation were in

excellent agreement with those observed by transmission electron microscope (TEM) images.

Raman spectroscopy in combination with chemometric analysis is used for discriminating early-

stage (stages 0 and I) esophageal cancer samples ex vivo. Partial least squares regression (PLSR)

analysis and self-organization maps (SOMs) well discriminated the datasets of cancerous and

normal samples. Linear discriminant analysis (LDA) based on Raman bands was able to predict the

tissue types with 81.0% sensitivity and 94.0% specificity.

References

1. K. Wongravee; T. Parnklang; P. Pienpinijtham; C. Lertvachirapaiboon; Y. Ozaki; C.

Thammacharoena and S. Ekgasit, Phys. Chem. Chem. Phys., 2013, 15, 4183.

2. T. Parnklang; C. Lertvachirapaiboon; P. Pienpinijtham; K. Wongravee; C. Thammacharoen; S.

Ekgasit, RSC advances, 2013, 3, 12886.

3. K. Nitinaivinij; T. Parnklang; C. Thammacharoen; S. Ekgasit and K. Wongravee, Analytical

Methods, 2014, 6, 9816

4. M. Ishigaki, Y. Maeda, A. Taketani, B.B. Andriana, R. Ishihara, K. Wongravee, Y. Ozaki, H.

Sato, Analyst, 2016, 14, 1027.


INV-38 84


Development of bacterial sensors based on conducting-polymer imprinting technology

Hiroshi Shiigi, and Tsutomu Nagaoka*

Department of Applied Chemistry, Osaka Prefecture University, Gakuen-cho, Naka-ku, Sakai,

Osaka 599-8570, Japan

*E-mail: nagaoka @ osakafu-u.ac.jp

Conducting polymers incorporate anionic substances in their textures to compensate for the cationic

charges arising during oxidative polymerization, and such substances can be removed from the

texture by overoxidation at high pH, leaving their complementary cavities behind [1].

In this paper, we report conducting-polymer based sensing systems for the application to bacterial

detection [2-4]. Bacterial species usually carry negative surface charges and are quite easily taken

up in conducting polymer films. Bacterial species, such as E. coli, are vertically inserted during

electrodeposition of polypyrrole films on metallic and carbon electrodes [2,4] and ejected after the

overoxidation of the films [3]. Thus, the uptake and ejection protocols can create cavities

complementary to the template bacteria.

The template bacteria were selectively bound to the imprinted films, compared to other different

bacterial species to demonstrate that this simple technique could be used as sensors for bacterial

detection.

Based on a similar protocol, gold nanoparticle/polyaniline nanocomposites were prepared for the

optical detection of bacteria [4]. As well as the imprinting technique, surface modification with

antibodies allowed us to add selectivity towards the targeted bacteria.

Thus, the present technique can be flexibly applied to films, microbeads, and nanocomposites for

the selective detection and incorporation of a wide range of bacterial species based on the

straightforward tailor-made protocols.

References

[1] S. Tokonami, H. Shiigi, and T. Nagaoka, in Molecularly Imprinted Sensors: Overview and

Applications, ed. S. Li, Y. G. Sergey, A. Piletsky and J. Lunec, 2012, Chap. 4, Elsevier B. V.

[2] S. Tokonami, K. Saimatsu, Y. Nakadoi, M. Furuta, H. Shiigi, and T. Nagaoka, Anal. Sci., 2012,

28, 319.

[3] S. Tokonami, Y. Nakadoi, M. Takahashi, M. Ikemizu, T. Kadoma, K. Saimatsu, L. Q. Dung, H.

Shiigi, and T. Nagaoka, Anal. Chem., 2013, 85, 4925.

[4] H. Shiigi, T. Kinoshita, M. Fukuda, D. Q. Le, T. Nishino, and T. Nagaoka, Anal. Chem., 2015,

87, 4042.

Keywords Bacterial detection; imprinting; polypyrrole; polyaniline; gold nanoparticle

INV-39 85


Radical Driven Peptide Sequencing Mass Spectrometry this is Close for Use in

Proteomics Applications

Han Bin Oh*

1Department of Chemistry, Sogang University, Seoul, 40002, Korea


Last decade or so, we have witnessed the development and an expanding use of electron-based

peptide tandem mass spectrometry methods such as electron capture/transfer dissociation (EC/TD).

Likewise, a variety of radical-driven peptide tandem mass spectrometry methods have been

developed towards its goal for applications in peptide/protein characterizations, even proteomics

applications. Along this line, our group developed the so-called TEMPO-assisted FRIPS (free

radical initiated peptide sequencing) mass spectrometry method: 2-(2,2,6,6-tetramethyl piperidine-

1-oxyl). This method is based on the extraordinary thermostability of TEMPO radical species. In

this method, a reagent o-TEMPO-Bz-C(O)-NHS is conjugated to the peptide N-terminus, or

sometimes lysine side chain if available, forming o-TEMPO-Bz-C(O)-Peptide. Upon collisional

activation, this conjugated peptide exclusively undergoes homolytic cleavage between the TEMPO

oxygen and benzyl carbon, generating •Bz-C(O)-peptide ions. The generated peptide radical ions

induce extensive peptide backbone dissociations, particularly exhibiting typical radical peptide

fragments such as a-/x- and c-/z-type fragments. However, this method has a few limitations in

order to be used in proteomics applications, for example, double-conjugation, solubility, two-step

activation issues. In recent years, these limitations are all overcome. In the symposium, the

limitations our TEMPO-based reagent faced and how these limitations could be overcome will be

presented. Furthermore, a few examples of how our method can be used in proteomics research will

also be introduced.

Keywords radical-driven peptide mass spectrometry; TEMPO; tandem mass spectrometry; FRIPS

(free radical initiated peptide sequencing)

INV-40 86


Direct Sample Analysis by Mass Spectrometry

Zhongping Yao

Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom,

Kowloon, Hong Kong

Email: [email protected].

Mass spectrometry (MS) is a powerful tool for qualitative and quantitative analysis of various samples,

and electrospray ionization (ESI) is a commonly used ionization techniques in mass spectrometry.

Conventional ESI-MS uses a capillary for sample loading and ionization, and typically requires sample

pretreatment and chromatographic separation prior to ESI-MS detection. In the past few years, our

group has developed electrospray ionization on various solid substrates, including inert materials such

as wooden tips, aluminum foil, pipette tips and TLC plates, solid samples such as tissues and bones,

and living organisms such as fish and toad. These techniques have allowed direct, real-time, in situ and

in vivo analysis of samples, and have been applied in various fields.

References:

1. Hu, B.; Yao, Z. P. Anal. Chem. 2016, 88, 5585-5589.

2. Hu, B.; Xin, G. Z.; So, P. K.; Yao, Z. P. J. Chromatogr. A 2015, 1415, 155-160.

3. Wang, H. H.; So, P. K.; Yao, Z. P. Anal. Chim. Acta 2014, 809, 109-116.

4. Hu, B.; Wang, L.; Ye, W. C.; Yao, Z. P. Sci. Rep. 2013, 3, 2104.

5. Hu, B.; Lai, Y. H.; So, P. K.; Chen, H. W.; Yao, Z. P. Analyst 2012, 137, 3613-3619.

6. Hu, B.; So, P. K.; Chen, H. W.; Yao, Z. P. Anal. Chem. 2011, 83, 8201-8207.

INV-41 87


Detection and mapping of trace materials on surfaces

under ambient conditions using multiphoton electron extraction spectroscopy (MEES)

Valery Bulatov, Anneli Kruve, Daniel Fisher, Shisong Tang, Nataly Vinerot, and Israel Schechter

Schulich Faculty of Chemistry, Technion - Israel Institute of Technology, Haifa 3200, Israel

[email protected]

Multiphoton electron extraction spectroscopy (MEES) is a new spectroscopy in which UV

laser pulses are utilized for extracting electrons from solid surfaces in multiphoton processes

under ambient conditions. Counting the emitted electrons as a function of laser wavelength

results in detailed spectral features, which can be used for material identification. The method

has been applied to detection of trace compounds on a variety of surfaces. It has been tested

for trace explosives, drugs, pesticides, PAHs, and many other materials. The analytical LODs

are in the sub pmole range, which indicates that MEES is one of the most sensitive detection

methods for solid surface under ambient conditions. Scanning the surface with the laser

allows for its imaging, such that explosives (as well as other materials) can be located. The

imaging mode is also useful in forensic applications, such as detection of drugs in human

fingerprints.

MEES imaging of a swab spiked with Semtex H (left) and the spectrum at a suspected point

(right).

1. S. Tang, N. Vinerot, D. Fisher, V. Bulatov, Y. Yavetz-Chen and I. Schechter,

Detection and mapping of trace explosives on surfaces under ambient conditions using

multiphoton electron extraction spectroscopy (MEES).

Talanta 155 (2016) 235–244.

2. S. Tang, N. Vinerot, V. Bulatov,Y. Yavetz-Chen and I. Schechter,

Multiphoton Electron Extraction Spectroscopy (MEES) and its comparison to other

spectroscopies for direct detection of solids under ambient conditions.

Anal. Bioanal. Chem., (2016) in press. Published on-line 1 Sept. 2016

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INV-42 88


Mobile Chemical Analysis (MCA) for Measuring Nutrient Substances by Computer-

Controlled Flow Chemical Analysis (CC-FCA) Techniques Using LED-Based Detectors

Lukman Hakim1, Yasutada Suzuki2 and Shoji Motomizu3

1 University of Brawijaya, Department of Chemistry, Science Faculty, Jl. Veteran, Malang,

East Java, Indonesia. 2 Yamanashi University, Interdisciplinary Graduate School of Medicine and Engineering, 4-4-37, Takeda, Kofu,

Yamanashi, 400-8510 Japan. 3 Okayama University, Incubator 109, 1-1-1 Tsushimanaka, Kitaku, Okayama, 700-8530 Japan.

[email protected]

A mobile chemical analysis (MCA) is often very useful and/or sometimes requisite in the area

concerning environmental issues. Such MCA techniques must be able to give high-quality and reliable

analytical results rapidly on site, and they must be easily handled and carried to any site of analyses;

the systems must be compact, portable, less heavy, robust and battery-driven. Further, analytical

methods for MCA must be robust, stable, simple and agile in instruments and techniques; less

complicated and less time-consuming techniques are desired.

In MCA, most of the techniques used are based on wet analysis; therefore solution handlings are

requisite and are very important. For accurate and reproducible solution handling, manual techniques

had better to be replaced with any computer-controlled fluid-flow chemical analysis (CC-FCA)

technique, such as FIA, SIA, MSFIA, SIC, AIA, SIEMA, Auto-Pret with a mini-column and SWIA,

where data acquisition/processing techniques can be used conveniently.

The determination of nutrient substances, such as nitrogen compounds (nitrate, nitrite and ammonia),

phosphorus (orthophosphate and other dissolved phosphorus compounds), silicate and potassium

present in macro amounts, and other minerals etc. in micro amounts in waters (river, lake, sea etc.) and

soils (fields, farms, gardens etc.) is very important in culturing fishes, plants etc. Such nutrient

substances, in macro and micro quantities, must be in adequate concentrations in waters and soils for

effective culture of fishes and plants. Too much nutrients (dissolved nitrogen and phosphorus

compound) in sea and lake can cause water eutrophication; sometimes it can cause red tide, which can

seriously damage fishes and plants in waters.

Usually, the monitoring of sea farms is carried out by sampling seawater manually on a ship every 2

or 3 hours, and analyzing them at a laboratory with some analytical instruments. Now any automated

monitoring systems for seawater on site are demanded eagerly in sea farms. Such monitoring systems

should automatically analyze water samples on site remotely for one or two weeks: the commands for

controlling the systems and analytical data obtained should be sent from/to a land station via

telecommunication devices.

For nutrient measurement, LED-based single and/or multi-channel absorptiometric and fluorometric

detectors can be favorably applied to MCA, which will improve the portability and agility of MCA

systems and techniques.

In the present study, the characteristics of CC-FCAs, such as SP (syringe propelling)-FIA, SIA, AIA

and SIEMA are compared with one another with respect to simplicity and robustness, sensitivity and

reproducibility of analytical results, agility and portability of the systems, and practical performance

when CC-FCA coupled with LED-based detectors are used for nutrient measurements.

Keywords Flow-based analysis; computer-controlled flow chemical analysis (CC-FCA), mobile

chemical analysis (MCA), nutrient substances, LED-light absorptiometric detector

INV-43 89


Development of simple analytical instruments for undergraduates and high school

students

Yasutada Suzuki

Department of Research Interdisciplinary Graduate School of Medicine and Engineering,

University of Yamanashi, Yamanashi, 4008510, Japan


Field portable analytical instruments have such virtues as compactness, robustness, low power

consumption, and so on. We have developed several portable instruments including light-emitting

diode (LED) based photometers, spectrophotometers and electrochemical analyzers. These features

are also attractive for educational use. A room for chemical experiments is often not large enough

for many students. Analytical instruments with small footprints can provide another space for

sample and reagent preparation.

At first, we have developed a photometric titrator and an absorbance detector for flow-injection

analysis based on an LED-based colorimeter and a power source for electrogravimetry and a

constant-current coulometric titrator for electrochemical analysis. We have designed these

instruments to reduce reagents and wastes, and some of them have been used for 10 years without

significant troubles at analytical chemistry experiments for undergraduates in University of

Yamanashi.

Then, we have developed kits of an RGB LED-based colorimeter and a potentiometer as education

materials for undergraduates and high school students. We have redesigned the previous

instruments in order to simplify the assemble process. Although wiring harnesses were prebuilt and

some surface mount ICs were soldered on a circuit board beforehand, most of electric parts,

including IC sockets, capacitors, resistors, and diodes were soldered by student themselves at

workshop or lecture. It took for about 2 hours to assemble each kit and check the function. This

colorimeter was successfully applied to the determination of iron in river water samples with 1,10-

phenanthroline and the potentiometer was used with a pH glass electrode. These instruments are

compact, but have sufficient performance for practical analyses.

Fig. 1 Photograph and schematic diagram of a portable colorimeter kit with an RGB LED as a light

source

Keywords educational material; student experiments; colorimeter; potentiometer.

INV-44 90


Construction of international collaboration for research and training of younger

generation scientists in flow based analysis

Tadao Sakai1*, Norio Teshima1, Shoji Motomizu2, Kate Grudpan3, Gary D. Christian4

1 Department of Applied Chemistry, Aichi Institute of Technology, Toyota 470-0392, Japan

2 Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan 3 Department of Chemistry, Faculty of Science, and Center of Excellence for Innovation on Analytical Science

and Technology, Chiang Mai University, Chiang Mai 50200, Thailand 4 Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA

The Japanese Association for Flow Injection Analysis (JAFIA) was founded on January 1st, 1984.

About 200 Japanese FIA researchers have joined and the first president of the JAFIA was Prof.

Nobuhiko Ishibashi of Kyushu University. He contributed until 1991 as the president. In the same

year, the first semi-annual meeting was held in Fukuoka on February 24th, 1984. And also, the

Journal of Flow Injection Analysis (JFIA, Vol.1, No.1) was published at the same time. Two issues

are published in June and December and Vol. 33, No.2 will be published on December in this year

In 1995, Gary D. Christian requested that JAFIA join to International Conference on Flow Injection

Analysis (ICFIA) to develop the JAFIA semi-annual meeting held in Japan and to exchange new

FIA technology because FIA and the related techniques are most important and effective in many

fields. The president of JAFIA was Prof. Shoji Motomizu for 10 years. The collaboration with Gary

and Shoji are wonderful for development of the joint conference. About 20 papers by Japanese

researchers were submitted in the joint conference held in Prague, 1999.

S. Motomizu and T. Sakai serve as steering committee members of the joint conference to enhance

the international activity. The conference is held in many countries and about 200 FIA researchers,

including students, join. JAFIA offers students excellent poster award at the ICFIA/JAFIA

conference.

Various meetings supported by the Thai Research Fund (TRF) are organized by the universities in

Chiang Mai (starting with Kate Grudpan’s grants), Bangkok and other cities. TRF with a variety of

grant mechanisms is important and useful for basic research, applied chemistry, development for

industry, Royal Golden Jubilee with Ph.D program, training younger generation,

international/national symposia, seminars, workshops and exchange among the researchers in next

generation. In 2015, the 108 TRF seminar was held in Chiang Mai University for encouragement of

faculty and students. About 100 people joined.

Okayama University and Chiang Mai University signed with MoU in 1999 and started the joint

research project. After then, Okayama University, Mahidol University and Chulalongkorn

University agreed to the exchange program for students and Faculty.

Aichi Institute of Technology signed with MoU between Chiang Mai University (in 2003), Prince

of Songkla University (in 2005) and Khon Kaen University (in 2011) for collaboration and

exchange programs on flow based techniques.

Keywords International collaboration; Training of younger generation ; Flow analysis,

fabrication of electrochemical /photoelectrochemical biosensor using novel...

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