Oral Presentation

Sample Pretreatment of Environmental Estrogens and Analytical Application Using
Liquid Chromatography-Mass Spectrometry
Jin-Ming Lin*, Wei Yan, Tianhe Jiang
Department of Chemistry, Tsinghua University, Beijing 100084, China
E-mail: jmlin@mail.tsinghua.edu.cn

In recent several decades, due to excessive use and discretional disposal of hormone pills andcontraception pills, increasing content of various estrogens have enter to the environmental waters. Since
these estrogens have ultra high physiology activity at very low concentration, they have become a threat to
eco-environment and human health. Therefore, Estrogens have aroused the attention of the world andbecome a new global environmental problem.
Because of high resolution and sensitivity, liquid chromatography-mass spectrometry (LC-MS) has
widely applied in environment, medical analysis, clinic detection and life science. Though LC-MS has
powerful analytical ability, when it is applied for real sample analysis, complex matrices often seriouslyinterfere the final results. In this case, to develop simple, rapid and effective pretreatment method is an
important impetus to broaden the application of LC-MS technology. The aim of this work is to develop
rapid and effective pretreatment method coupled with LC-MS for quantitative analysis of estrogens invarious matrices. 1) Ten highly potent estrogens were identified and quantified by solid-phase extraction(SPE) followed by LC-MS. Improved two-step SPE process was employed in the work. C18 cartridge wasused for both enrichment of all target estrogens and retention of some nonpolar impurities, and then a polar
Florisil cartridge was subsequently used to separate the interested estrogens from the polar impurities. In
the optimized condition, this method was used to verify the presence of the target analytes in Qinghe Riverand influent of sewage treatment plant (STP) in northwest of Beijing, China. Some estrogens were detectedin the river water samples and sewage water samples at relatively high concentrations. The developed
method proved to be effective for analyzing estrogen compounds in complex matrices. Moreover, theachieved results demonstrated that a lot of concern should be addressed about the potential risk of thepresent of estrogens in aquatic environment. 2) Triacontyl bonded silica (C30) material was applied as SPE
sorbent and a SPE-LC-MS method was established for the determination of 8 estrogens and their
metabolites in water samples. The results showed superior performance of C30 to C18 by the shorter
treatment time and fewer required elution solvent. Method validation using C30 packed cartridge was also
testified with spiked real water samples, including tap water and river water. 3) Using C30 as on-line SPEmaterial and a specially designed online analytical system which allowed large sample volume injection, ahigh speed and robust online SPE-HPLC-MS method was established for the analysis of five estrogens andbisphenol A (BPA) in milk samples.

Biography
Professor Jin-Ming Lin (Ph.D)
Professor of Cheung Kung Scholarship, Ministry of Education, China
Vice Chair, Department of Chemistry, Tsinghua University, Beijing 100084, China
Tel/Fax: +86-10-62792343
E-mail: jmlin@mail.tsinghua.edu.cn

Professor Jin-Ming Lin was born in 1963 in Fujian, China. He received his BA degree in 1984
and received a PhD degree in analytical chemistry at Tokyo Metropolitan University in 1997.
He had studied and worked in Showa University and Tokyo Metropolitan University during
1992-2002. He is currently a full professor of Tsinghua University since 2004. He is a vicechairman of the Division of Analytical Chemistry, Chinese Chemical Society, and a memberof council of Chinese Society for Chromatography Science. He is a contributing editor ofTrends in Analytical Chemistry, a guest editor of Talanta for special issues ofChina-Japan-Korea Environmental Analytical Chemistry and editorial broads of Talanta,
Luminescence, Eurasian Journal of Analytical Chemistry, J. Flow Injection Analysis,
International Journal of Analytical Chemistry and seven Chinese journals. He received several
awards for his contributions in chemiluminescence and separation science: Outstanding
Young Chemist Award (Chinese Chemical Society, 1992), Young Analyst Award for Flow
Injection Analysis (Flow Injection Section, Japan Society of Analytical Chemistry, 2000) ,Kanton New Century Award (Japan Society of Analytical Chemistry, 2001), National
Science Fund for Distinguished Young Scholars of China (National Natural Science
Foundation of China, 2002), FIA Award for Science (Flow Injection Section, Japan Society ofAnalytical Chemistry, 2008) and GC Contribution Award (GC Discussion Group, Japan
Society of Analytical Chemistry, 2008)
His current research is focused on sample pretreatment, chemiluminescence, capillaryelectrophoresis and microfluidic device. He is the author and co-author of 218 original
research papers published in international journals, 21 reviews, 3 books and 31 patents.

Arsenic speciation by coupling liquid chromatography
with gradient hydride generation atomic absorption
spectrometry
Yong Tiana, Ming-Li Chena, Xu-Wei Chena, Jian-Hua Wanga*aResearch Center for Analytical Sciences, Northeastern University, Box 332,
Shenyang 110004, China
Yoshihiro Hiranob, Hideyuki Sakamotob, Ikumei Setsubb

Hitachi High-Technologies Corporation, Hitachinaka, Ibaraki, 312-8504, JapanThe accurate determination of arsenic in various sample matrices has attracted
extensive attentions because of its high toxicity. The total concentration of arsenic is
not sufficient to provide comprehensive information about the toxic effect, while itdepends strongly on the chemical state of a specific arsenic species. Therefore, theidentification of arsenic species is one of the most important issues when evaluating
its toxic effects [1]. The hyphenation of liquid chromatography with hydride generation
atomic spectrometry is among the most common techniques for the separation andquantification of metal species [2]. The present work focuses on the development of asuitable procedure for arsenic speciation by coupling liquid chromatography withgradient hydride generation atomic absorption spectrometry (HPLC-GHG-AAS).
Fig. 1 illustrated the HPLC-GHG-AAS system for arsenic speciation. The arsenic
species were separated by the HPLC system, followed by mixing with reagents HCl
and NaBH4 to facilitate hydride generation in a reaction coil in a boiling water bath.
The reaction mixture was cooled off in a cooling water bath followed by separation of
hydride in a gas-liquid separator, which was afterwards quantified by a flame AAS(FAAS). The boiling water bath was used for improving the efficiency of hydride
generation of arsenic species, while the cooling water bath eliminates any entrapmentof water vapor into the argon stream and the quartz cell of FAAS.

Fig.1. The HPLC-GHG-AAS system for arsenic speciation.
The separation of arsenic species was performed by using a C30 column at 25℃.
The mobile phase contains 5 mM of sodium butanesulfonate, 4 mM of malonic acid, 4
mM of tetramethylammonium hydroxide and 0.1% methanol (pH=2). The mobile
phase flow rate is 1.0 mL/min. The gradient hydride generation is performed such thata 6 M of hydrochloric acid was used for the hydride generation of arsenate, arsenite
and MMA, when the signal for MMA has been recorded, the controlling valve wasswitched to introduce a stream of 0.6 mol/L of hydrochloric acid as hydride
generation medium for the other two arsenic species, i.e., DMA and TMA (Fig. 2).
The detection limits for the arsenic species were summarized in Table 1.
Ultrasonic-assisted extraction was used for the pretreatment of Hijiki, i.e., 5 gramof Hijiki was taken into a 100 mL vessel to which 50 mL water was added. The
mixture was sonicated for 30 min and the extract was separated by filtration forarsenic speciation by appropriate dilution. Fig. 3 showed the analytical results from a
5-foild diluted extract of Hijiki. Three arsenic species, i.e., arsenate, DMA and TMA,
were identified in Hijiki samples.
0.9

Hijiki 1

As(V)

0.09

AbsorbanceAbsorbance

0.7
0.04

0.5
-0.01
350

0.3

550

DMA

0.1

750

TMA

-0.1
0

100

200

300

400

500

600

700

800

0.9

Hijiki 2As(V)Absorbance

0.7

C

6.0M & 0.6M HCl

B

6.0M HCl

A

0.6M HCl

Time / s

0.09

0.04

0.5
-0.01
350

0.3

550

DMA

0.1

750

TMA

-0.1
0

100

200

300

400

500

600

700

800

Time / sFig. 2. Arsenic speciation in aqueous
solution by the HPLC-GHG-AAS system

Fig. 3. Arsenic speciation in 5-folddiluted Hijiki extract

Table 1. The speciation of arsenic in Hijiki by the HPLC-HG-AAS system
SpeciesFound
Hijiki 1 (µg/g)

Found
Hijiki 2 (µg/g)

Spiked(µg/L)

Spiking Recov.
Hijiki 1 (%)

Spiking Recov.
Hijiki 2 (%)As(V)

53.6±9.2

60.8±13.2

50

107.4

111.2

As(III)

n.d.

n.d.

50

97.9

97.1

MMA

n.d.

n.d.

50

106.9

104.9

DMA

2.7±0.4

2.4±0.5

50

112.5

105.7

TMA

0.34±0.12

0.15±0.03

50

104.6

102.2

1. Yan, X.-P.; Yin, X.-B.; He, X.-W.; Jiang, Y. Anal. Chem. 2002, 74, 2162-2166.
2. Mir, K.A.; Rutter, A.; Koch, I.; Smith, P.; Reimer, K.J.; Poland, J.S. Talanta 2007,
72, 1507-1518.

Introduction to the author:
Name:

Jian-hua Wang

Date of birth:

Nov. 04, 1962

Nationality:

ChineseAffiliation:

Research Center for Analytical Sciences, Northeastern University,
Shenyang 110004, China
Tel: 86-24-83688944, E-mail: jianhuajrz@mail.neu.edu.cn

Title:

Professor of Analytical Chemistry

J.-H Wang obtained his B.Sc from Department of Chemistry, Nankai University, and
M.Sc from Jilin University, China. He was awarded a PhD degree by Technical
University of Denmark. He has been an Associate Editor for TALANTA since Dec.
2004. He is currently a member of International Advisory Board for Journal of
Analytical Atomic Spectrometry.
His research interest is focused on the exploitation of flow analysis protocols andhyphenation techniques, isolation of trace level of analytes from complex sample
matrices and life science analysis.

Polydimethylsiloxane Mini-Disk Extraction as a Novel Sampling Technique
Eun-Ju Cha, Dong-Sun Lee*dslee@swu.ac.kr
Department of Chemistry, Seoul Women’s University 139-774 Korea
A novel sampling method called headspace polydimethyl-siloxane (PDMS) mini-disk
extraction (HS-PDE) was developed, optimized, validated and applied for the GC/MS
analysis of flavors for the first time. A prototype PDMS mini-disk (diameter: 8 mm,
thickness: 0.157 mm, weight: 9.4 mg) has been designed as a sorption device. The
technique uses a small PDMS mini-disk and very small volume of organic solvent. This
new HS-PDE method is very simple to use, inexpensive, rapid, requires small sample
amounts and solvent consumption. Because the acetonitrile for extraction is reduced to avery small volume of only 0.03 mL, there is minimal waste or exposure to toxic organic
solvent and no further concentration step. Linearities of calibration curves for α–pinene,
β–pinene, limonene and γ–terpinene by HS-PDE combined with GC/MS were generally
good. LOD and LOQ showed very low values, and dynamic ranges were 6.00 ng~3500
ng. This method exhibited good precision and accuracy. The overall extraction
efficiency of this new method was evaluated by using partition coefficients (Kfg) andconcentration factors (CF) for several characteristic components from nutmeg and mace.
Partition coefficients were in the range from 2.04 x 104 to 4.42 x 105, while CF values
were 0.88~15.03. HS-PDE was applied successfully for the analysis of flavors
compositions from cumin, nutmeg and mace. The HS-PDE method is a very promisingsampling technique for the characterization of volatile flavors.

Biography
Name: Dong-Sun Lee
Affiliation: Department of Chemistry, Seoul Women’s University
Tel: +82(2)970-5654 e-mail: dslee@swu.ac.kr
URL: http://mail.swu.ac.kr
Title: Dean of Graduate School, Professor
Professor Dong-Sun Lee received his Ph.D. degree in analytical chemistry in 1984 from
Sung Kyun Kwan University in Seoul. In 1985, Professor Lee joined the chemistry
faculty at Seoul Women’s University. He spent one year sabbatical work at the
Washington State University of U.S.A. in 1995. He received Academic Award in 2000
from Analytical Chemistry Division of The Korean Chemical Society. Professor Lee
served in the Korean Chemical Society as the Chairman of the Analytical Chemistry
Division, and as Vice-chair in the Korean Society of Analytical Sciences in 2002. He is
now serving as a Dean of Graduate School of Seoul Women’s University from 2008. He
has a wide variety of interests in the fields of analytical separation using GC/MS,
LC/MS and ion mobility spectrometry. His current research is mainly focused on
analytical chemistry of natural aroma compounds. He has more than 160 publications,edited books and patents.
------------------------------Name: Hye-Jin Jang
Affiliation: Department of Chemistry, Seoul Women’s University
Tel: +82(2)970-7712
e-mail: 01199377332@hanmail.net
Position: Graduate student (M.S.), Teaching Assistant------------------------------Name: Hyun-Hwa Son
Affiliation: Department of Chemistry, Seoul Women’s University
Tel: +82(2)970-7712 e-mail: balloon0502@hanmail.net
Position: Undergraduate student

Design of Flow Injection Analysis to Realize Green Chemistry
T. Takayanagi1
1Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushimanaka, Kita-ku,

Okayama City, Okayama Prefecture 700-8530 Japan
Summary
A sense of Green Sustainable Chemistry will be incorporated in analytical chemistry to perform advanced
analytical systems. Use of conventional instruments and common reagents will be alternatives to theanalytical instruments consuming large amounts of energy and natural resources. Flow injection analysisof aluminum with fluorimetric detection and sequential injection analysis of iron with chemiluminometricdetection are introduced as examples.
Keywords: green sustainable chemistry, low consumption of energy and resources, conventional
instruments, common reagents, alternative analytical systems
1 Introduction
Under limited energy and natural resources on earth, sciences are now directed toward
environment-friendly systems: less consumption of energy and resources, reuse and recycle of materials,
zero emission of pollutants and wastes. These actions are generally called as Green Sustainable Chemistry.
The concept is also valid in the field of analytical chemistry, deeply related with environmental sciences.
During last century, highly sensitive analyses have been established by developing high-performance
analytical instruments such as ICP-AES. Analysis by ICP instruments are realized based on the
consumption of a large amount of noble gases. Therefore, interests nowadays are directed toward how we
can realize the sensitive analysis by convenient instruments with common reagents. Although theanalytical systems thus established are based on inexpensive items, it would be highly integrated andsophisticated analytical systems.
Development of new analytical reagents has also been one of major trends in analytical chemistry.
So-called “new” reagents sometimes realized high sensitivity and high selectivity. On the other hand, the“new” reagents simultaneously possess the risk of toxicity/pollution. History of useful/convenientsynthesized substances tells us that they are also the one of toxicity/pollution. Analytical chemists should
weigh the benefits with the possible disadvantages. The conventional instruments and common reagentswill be utilized to establish the analytical systems and to meet Green Sustainable Chemistry. Flow
injection analysis and sequential injection analysis systems are also helpful to reduce the chemical wastes.

Corresponding author, T. Takayanagi, takayana@cc.okayama-u.ac.jp, 3-1-1 Tsushimanaka, Kita-ku
Okayama 700-8530

2 Determination of aluminum with 8-quinolinol by fluorophotometric flow injection analysis
Derivatizing reagent of 8-quinolinol is well known for the determination of several metal ions byfluorophotometry. Solvent extraction with organic solvents are usually utilized together to improve thefluorescence intensity and the extraction concentration. The organic solvents are volatile and they would
be reduced. Micelle sensitization was utilized to improve the solubility of the complexes and to improve
the sensitivity in an aqueous solution; nonionic Triton X-100 was used as a micelle-forming surfactant.
By coupling with flow injection analysis, limit of detection at 3x10−9 M was attained for aluminum, andthe analysis method was applied to river water samples. The FIA signal increased with time after the
addition of 1 mM HNO3, and the profiles suggested the decomposition of complexed aluminum species in
an aqueous solution.
3 Chemiluminometric detection with luminol in sequential injection analysis
Luminol is a well-known and commonly-used reagent for the chemiluminometric detection of H2O2 orcatalytic metal ions. Detection method of chemiluminescence is advantageous in sequential injection
analysis, because the schlieren effect, when auxiliary light is used, will not occur and ghost signal is much
reduced. The flat baseline realizes the high sensitive detection. Micelle sensitization on Fe3+/Fe2+
catalyzed luminol chemiluminescence was examined. Nonionic surfactant of Triton X-100 was added in
the metal ion solution, luminol in weakly alkaline solution, or H2O2 solution. Among them, addition in themetal ion solution much improved the chemiluminescence intensity about 13 times, which will contributeto develop highly sensitive determination of the metal species.
4 Conclusion
In summary, design of analytical systems using conventional detections and common reagents will
replace high-performance analytical instruments. The consumption of energy and resources will be
reduced, and suspicious novel reagents will be excluded, which lead us to Green Chemistry.
5 Biographical Sketch
Name : Toshio TAKAYANAGI
Born : Saitama, Japan
Nationality : Japan
Affiliation : Okayama University
Contact : E-mail: takayana@cc.okayama-u.ac.jp
Web: http://chem1.chem.okayama-u.ac.jp/~separation/taka_j.html
Title : Associate Professor
Record : Encouragement award from The Society for Chromatography Sciences, Japan (2004).

Corresponding author, T. Takayanagi, takayana@cc.okayama-u.ac.jp, 3-1-1 Tsushimanaka, Kita-ku
Okayama 700-8530

Hybrid organic-inorganic silica monolith with hydrophobic /strong cation-exchange
functional groups as a novel sorbent for in-tube solid-phase microextraction
Ming-Ming Zheng, Ge-Deng Ruan, Yu-Qi Feng *Key laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education),
Department of Chemistry, Wuhan University, Wuhan 430072, China
E-mail address: yqfeng@whu.edu.cn
Sample treatment has been recognized as the main bottleneck of the analytical process, especiallywhen trace analysis in complex matrices is the purpose. As to the development of environmentally andbiologically favorable analytical methods, miniaturization trends of the sample preparation with low sample
requirements, and low solvent consumption has been gained much attention. In-tube solid-phasemicroextraction (in-tube SPME) was introduced as an excellent sample preparation technique, since it
possesses several attractive features including small sample volume, simplicity and easy automation.
For in-tube SPME, the sorbent in a capillary play an important role in interference elimination andeluting the analytes more selectively. Therefore, developing sorbent with high extraction efficiency andhigh selectivity for in-tube SPME is always an attractive task. In this contribution, we prepared a novel
hybrid organic-inorganic silica monolith with hydrophobic and strong cation-exchange functional groups ina fused silica capillary and used it as a sorbent for in-tube SPME. The hybrid silica monolith functionalizedwith octyl and thiol groups was conveniently synthesized by hydrolysis and polycondensation of a mixtureof tetraethoxysilane (TEOS), n-octyltriethoxysilane (C8-TEOS) and 3-mercaptopropyltrimethoxysilane(MPTMS) via a two-step catalytic sol-gel process. Due to the favorable chemical reactivity of mercapto
pendant moieties, the obtained hybrid monolith was oxidized using hydrogen peroxide (30% w/w) to yield
sulfonic acid groups, which provided strong cation-exchange sites. The obtained hybrid monolith wascharacterized by diffused infrared spectroscopy, elemental analysis and scanning electron microscopy. The
results show that the resulting monolith contains much higher carbon (31.6%) and sulfur (4.8%) contents
than traditionally bonded silica materials. The extraction performance of the hybrid monolith was evaluatedusing sulfonamides as model analytes by in-tube SPME on-line coupled to HPLC. The results show that the
hybrid monolith with hydrophobic and strong cation-exchange functional groups exhibits high extraction
efficiency towards the model analytes with satisfactory reproducibility and stability. Finally, a method forthe determination of sulfonamide residues in milk was developed by the combination the hybrid monolithin-tube SPME and HPLC.

Keywords: In-tube solid-phase microextraction; Organic-inorganic hybrid materials; Silica monolith;
Mixed-mode; Hydrophobic interaction/cation-exchange; Sulfonamides

Biography
Professor Dr. Yu-Qi Feng
Director of the Research Center for Analytical Science,
Department of Chemistry, Wuhan University, Wuhan 430072, China.
Tel/Fax: +86-27-68755595
Email: yqfeng@whu.edu.cn

Dr. Yu-Qi Feng was born in 1962 in Fujian, China. He received his BA degree and MS degreefrom Lanzhou University in 1982 and 1985, respectively, and spent the next six years as an
Assistant Professor and then Lecturer in Department of Chemistry, Central China Normal
University. In 1991, he had the honor to obtain a national scholarship from the Ministry of
Education, Japan, and studied as a research student in Department of Chemistry at Ibaraki
University, Japan. In 1993, he moved to Chiba University in Japan to undertake a Ph. D.
degree and completed it in 1996. After two-year postdoctoral training at Wuhan University, hejoined this university as an associate professor in Department of Chemistry in 1998. Twoyears later, he was promoted to a full professor. He was a deputy dean of College ofChemistry and Molecular Sciences at Wuhan University from 2001 to 2004. From July to
November of 2002, he was working as a senior visiting scholar at Iowa State University andUniversity of California at Riverside, respectively.
His research interests are on the development of novel separation media for liquid
chromatography, capillary electrochromatography, capillary electrophoresis, and solid-phase(micro) extraction, and their applications to biomedical analysis and food analysis.

High extraction efficiency for polar aromatic compounds in nature water samples using
multiwalled carbon nanotubes/Nafion solid-phase microextraction coatings
Wen-Feng Chena, Jing-Bin Zenga, Jin-Mei Chena, Yi-Ru Wanga, Xi Chena,b*aDepartment of Chemistry and the Key Laboratory of Analytical Sciences of the Ministry of Education,
College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, Chinab
State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China

Since solid-phase microextraction (SPME) was introduced by Arthur and Pawliszyn in 1990, it has been
widely applied in many areas. Compared to traditional techniques, SPME offers many advantages such as highsensitivity, reproducibility and solventless and combines sampling, extraction, and sample introduction into asingle step. This technique is based on the partitioning of analytes between the sample and the coating of
SPME fiber. In order to achieve higher applicability for various compounds, various approaches including
physical deposition, electrochemical procedures, direct use of uncoated fiber, epoxy-glued solid sorbents,
sol–gel technology, and molecular imprinting polymers (MIPs), have been developed for the production of
SPME fibers. Nafion is a sulfonated tetrafluoroethylene based fluoropolymer-copolymer. The combination of
the stable Teflon backbone with acidic sulfonic group gives Nafion excellent characteristics such as highly
conductive, high operating temperature and chemical stability. In 1998, Nafion was applied as a fiber coating
material for SPME. It was used to extract alcohols from hexane and gasoline samples, and exhibited better
extraction efficiency than all of the commercially available coatings selected. These results indicate the great
analytical potential of Nafion to be an effective solid-phase microextraction coating for polar compounds.
In this study, a novel solid-phase microextraction (SPME) fiber coated with multiwalled carbon nanotubes(MWCNTs)/Nafion was developed and applied for the extraction of polar aromatic compounds (PACs) inenvironmental water samples. The characteristics and the application of this fiber were investigated. Electron
microscope photographs indicated that the MWCNTs/Nafion coating with average thickness of 12.5 µm washomogeneous and porous. The MWCNTs/Nafion coated fibers exhibited higher extraction efficiency towards
polar aromatic compounds compared to an 85 μm commercial PA fiber. SPME experimental conditions, suchas fiber coating, extraction time, stirring rate, desorption temperature and desorption time, were optimized inorder to improve the extraction efficiency. The calibration curves were linear from 0.01 to 10 µg mL−1 for five
PACs studied except p-nitroaniline (from 0.005 to 10 µg mL−1 ) and m-cresol (from 0.001 to 10 µg mL−1), anddetection limits were within the range of 0.03 to 0.57 ng mL-1. Single fiber and fiber-to-fiber reproducibility
were less than 7.5 (n=7) and 10.0 % (n=5), respectively. The recovery of the PACs spiked in nature water
samples at 1 µg mL−1 ranged from 83.3 to 106.0 %.

Biography
Professor Dr. Xi Chen
Department of Chemistry and Key Laboratory of Analytical Sciences of the Ministry of Education, College ofChemistry and Chemical Engineering, Xiamen University
422 Si-Ming South Road, Xiamen, Fujian, China 361005
Tel&Fax: +86-592-2184530
E-mail: xichen@xmu.edu.cn

Professor. Xi Chen, born in 1964, graduated from Fuzhou University with a Bachelor’s
degree of analytical chemistry, went on to obtain his Master’s degree there. Later in 1991,
awarded the Monbusho Scholarship from Japanese government, he entered Kyoto Institute ofTechnology to go on with his research and received the Ph.D degree four years later. During his
stay in Japan, he committed himself in the research fields on the application of photosensitive
materials related ruthenium in analytical chemistry.
Between the year of 1996 and 1998, he performed as a postdoctoral researcher at Xiamen
University, sooner after which, he was promoted as an associate professor at the department ofchemistry in that University. In 2003, he was promoted to be a full professor at the samedepartment.
Between June to July, 2002 and 2003, he was a senior research staying at Department ofChemistry, University of Hong Kong. He got JSPS finial support to collaborate with Prof. M.
Oyama, Kyoto University, in 2004.
At present, he is taking charge of a National High Technical Development Project (863 Project)Foundation in the field of ocean, National Scientific Foundation of China and a Key Project of
Fujian Scientific Foundation on the determination of pesticide residue. Dr. Chen is now
interested in chemiluminescence, separation analysis, chemical and biosensors.
In 2001, Dr. Chen was honored the Outstanding Contribution Award for his excellent work in
the 818 Subject of 863 Project and received the 1st and 3rd class Scientific Progress Prizes of
Fujian province in 2002, 2005 and 2007.

Application of Flow-Based Analysis to Diabetes Care
Norio Teshima and Tadao Sakai
Department of Applied Chemistry, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho, Toyota
470-0392, Japan
Summary
Vanadium has recently attracted much attention for its potential role against diabetes. Bottled drinking
waters containing vanadium are now commercially available. An automated stopped-in-dual-loop flow
analysis (SIDL-FA) system is proposed for the determination of trace vanadium in drinking water
samples. On the other hand, measurement of albumin/creatinine ratio is useful for detection of early-stage
diabetic nephropathy. In the present paper an alternative method for the successive determination ofurinary albumin and creatinine by sequential injection analysis is also described. We discuss here theapplicability of flow-based analyses to diabetes care.
Key word: Vanadium; Albumin; Creatinine; Flow based analysis; Spectrophotometry; Diabetes.
1 Introduction
In Japan approximately 38% of patients who started undergoing artificial dialysis in 2001 were due totheir diabetic nephropathy [1]. Therefore detection of early-stage diabetic nephropathy is important.
Heyliger et al. reported that vanadate (vanadium in pentavalent state) appeared to have an insulin-likeaction in the in vivo experiments on diabetic rats [2]. Since then, many studies have been reported on theinsulin-like effect of vanadium. Some of bottled drinking waters containing naturally occurring vanadium
are now available for health benefit. However, the exact role of vanadium has yet to be established.
Therefore the concentration of vanadium in such bottled waters must be strictly controlled.
Microalbuminuria is an indicator of early-stage diabetic nephropathy and can be measured using a 24
hour urine specimen. However, this method has difficulty for assessing microalbuminuria due tovariations in protein concentration over time. To eliminate the uncertainty and the strain, a moreconvenient method to detect microalbuminuria is the albumin/creatinine ratio (ACR) measured in arandom urine specimen.

2 Stopped-in-Dual-Loop Flow Analysis (SIDL-FA)
of Vanadium [3,4]The chemistry is based on the vanadium-catalyzed
oxidation reaction of p-anisidine by bromate in thepresence of Tiron as an activator to produce a dye(l max = 510 nm). The SIDL-FA system is shown in
Fig. 1. A 100-μL coiled loop around a heated device
is fitted onto each six-way injection valve. A
well-mixed solution containing reagents and

L1
B
1

S1
2

V1
4

L2
H

TSC

H

V2

V3

3
S2
RS1
RS2
RS3

P1
P2
P3

B
BP
D
ON
OFF

W

R
PC

Fig. 1 Schematic flow diagram of the SIDL-FA system. B,
HNO3; S1, and S2, sample; RS1, mixture of p-ansidine andacetate buffer (pH 3.3); RS2, KBrO3; RS3, Tiron; V1,
selection valve; V2 and V3, injection valves; P1, P2 and P3,
pumps; H, heater (105°C); L1, first loop; L2, second loop;
D, spectrophotometer (510 nm); R, recorder; PC,
computer; BP, back pressure restrictor.

Corresponding author, Norio Teshima, E-mail teshima@aitech.ac.jp

standard/sample is loaded into the first loop on a six-way valve, and then the same solution is loaded
into the second loop on another six-way valve. The solutions are isolated by switching these two
six-way valves, so that the catalytic reaction can be promoted. Then each resulting solution isdispensed to the detector with suitable time lag. The proposed SIDL-FA method allowed vanadium
to be quantified in the range of 0.1 to 2 μg L–1 and was applied to the determination of vanadium indrinking water samples.

In this work, two specific reactions were used

A

1000

Creatinine

y = 0.939x + 10.0
r2 = 0.989

800
600
400
200
0
0

250

500

750

1000

1250

Conc. of protein reported by Bradford protein
assay / mg dL-1 [32]separately to determine albumin and creatinine

texp 0.9829 < tcritical 2.024
38 degrees of freedom
95% confidence level

levels. First, eosin Y was employed to form a color

Conc. pf creatinine by proposed
method/mg dL-1

and Creatinine [5]Conc. of protein by proposed
method / mg dL-1

3 Sequential Injection Analysis (SIA) of Albumin

Albumin
1200

180
160
140
120
100
80
60
40
20
0

y = 1.02x + 0.57
r2 = 0.976

B

0

50

100

150

200

Conc. of creatinine reported by creatinine emzymatic
assay /mg dL-1 [33-34]

texp 0.5378 < tcritical 2.024
38 degrees of freedom
95% confidence level

2 Comparisons of two measurement
complex with albumin. The concentration of this Fig.
methods, successive determination of albumincomplex could be monitored by absorption at 547 and creatinine using the proposed SIA system
nm. Second, creatinine levels were determined by

and Bradford protein assay (A) and creatinine
enzymatic assay (B).

Jaffe’s reaction, where creatinine quantitatively produces an orange color with sodium picrate inalkaline medium. The absorbance was measured at 500 nm. These two detection reactions wereintroduced into a SIA system. The proposed method was applied to successively evaluate albumin
and creatinine in urinary samples taken from diabetic individuals. The results obtained by the
proposed method were good agreement with those obtained by official methods (Fig. 2).
4 References[1] S. Hara, Nippon Jinzo Gakkaishi 45 (2003) 65[2] C.E. Heyliger, A.G. Tahiliani, J.H. McNeill, Nature 227 (1985) 1474.
[3] N. Teshima, S. Ohno, T. Sakai, Anal. Sci. 23 (2007) 1.
[4] N. Teshima, M. Kuno, M. Ueda, H. Ueda, S. Ohno, T. Sakai, Talanta, in press.
[5] W. Siangproh, N. Teshima, T. Sakai, S. Katoh, O. Chailapakul, Talanta, in press.
5 Biographical Sketch
Name : Norio Teshima; Born : Okaya, Nagano; Nationality : Japanese
Affiliation : Aichi Institute of Technology
Contact : E-mail: teshima@aitech.ac.jp; Web: http://aitech.ac.jp/~analabo/
Title : Associate Professor, Dr.
Record : D.Sc. awarded by University of Tsukuba, Mar. 1996
Apr. 1996 to Mar 1997, worked as a research postdoctoral fellow of the JSPS.
Apr. 1997 to Mar. 1998, worked as an Assistant Professor at University of Tsukuba
Apr. 1998 to Mar. 2001, worked as a Lecturer at Aichi Institute of Technology
Apr. 2001 to present, worked as an Associate Professor at Aichi Institute of Technology
Aug. 2003 to Jul. 2004, studied abroad as a postdoctoral fellow at Texas Tech University, USA

New approach of simple pretreatment for GCMS analysis
by using monolithic material sorption extraction
for determination of 16 PAHs
Yuki Hashi1*, Manami Takeda2, Jin-Qiang Zhang1, Jun Fan1, Yoshihiro Nagami2, Atsushi Sato2
1

Shimadzu International Trading (Shanghai) Co. Limited

2

GL Science Inc.
E-mail: y-hashi@shimadzu.co.jp

Environmental pollutions and food contaminations are biggest social concerns, nowadays. Because of
the complex matrices in environmental substances and food stuffs, sample preparations for the quantitative
analysis of toxic and hazardous compounds with chromatographic instruments are inevitable in order toobtain reliable results. Gas chromatography mass spectrometry (GCMS) has become the important meansin the areas of qualitative and quantitative analysis of environmental monitoring and trace amount ofcontaminants in food stuffs. Although the performances of sensitivity and selectivity of this instrument
have rapidly been improved in recent years, the sample pretreatment procedures remain the bottleneck for
rapid analysis. Therefore, economical and effective pretreatment methods to eliminate matrix interferences,to achieve high throughout and to enrich the analyte for high sensitive detection are demanded.
The newly developed monolithic material sorption extraction (MMSE) has several advantages such as
effective pre-concentration and cleanup of organic molecules from gaseous and liquid samples, and simple
operation. Since monolithic material has wide through pore and meso pore structure, rapid permeability oforganic molecules and high trap efficiency with large surface area can be realized. The thermal desorption
technique together with GCMS measurement is most convenient for desorption of the analyte from themonolithic material.
MonoTrapTM (GL Science Inc, Japan) as a MMSE has been applied for the determination of 16
polycyclic aromatic hydrocarbons (PAHs) in water. After one hour extraction, MonoTrapTM device wastransferred to OPTIC3 thermal desorption device (ATAS GL International BV) which was accommodatedin GCMS-QP2010 Plus instrument (Shimadzu Corporation, Japan), and then GCMS analysis was carriedout. All the analytical performances have been evaluated. PAH (Fluoranthene, Benzo[b]fluoranthene,
Benzo[k]fluoranthene,Benzo[a]pyrene) recoveries were higher than 85% for spiked water sample. Therewas good linear correlation between peak area and PAH concentration with a linear range of 0.1–100
ng/mL and correlation coefficients >0.995. The repeatability (RSD) at 100 ppb concentration was less than
6～15 %.
Thermal desorption GCMS combined with MMSE has been confirmed as a simple pretreatment methodfor determination of 16 PAHs.

Biographical Sketch
Name

Yuki Hashi, (Ph.D.)

Title

Shimadzu International Trading (Shanghai) Co., Limited
Shimadzu Shimadzu Global COE for Application & Technical Development
General Manager, Chief Scientist

Address Shimadzu International Trading (Shanghai) Co., Limited , Shanghai Office
Block E, No.570 West Huaihai Road, 200052, Shanghai, China
Phone: +86-10-8525-2352
Fax: +86-10-8525-2330
E-mail: y-hashi@shimadzu.co.jp
http://www.shimadzu.net.cn
Birthday27th February, 1961

Nationality Japan
Research Major
Dr. Hashi graduated from Dept. of Chemistry, Osaka University, Osaka, Japan in 1986. At thesame year, he joined Shimadzu Corporation, analytical instrument division to develop HPLC
instrument especially electrochemical detector and column packing materials for protein
analyses. In 1994, he moved to Shimadzu Singapore as an application manager for
chromatography instruments to take care of the customers in South-East Asia as well as south
Asia countries. His major area of interest is development of new application system using LC,
GC, LCMS and GCMS for the enhancement of analysis efficiency. Based on this researchwork, he has received a Ph.D. degree from Research Center for Eco-Environmental Sciences,
The Chinese Academy of Sciences in 2009. Currently, he is interested in themulti-dimensional GC/GCMS system for expansion of applicability. Application area is not
only for food safety but also, for pharmaceutical, environmental and life science such as
genomics and proteomics. Beside his research work, he is also actively providing the lectureof basic instrumentation in university, and contributing his papers to some technicalconferences.

Analytical Method of Trans Fats by Comprehensive GCxGC and NMR
Jaeho HA*, Miyoung YOO and Dongweon SEO
Korea Food Research Institute, Republic of KOREA
Summary
The analytical condition of comprehensive GC×GC for TFA analysis has been optimized. Primary andsecondary column was sp-2560(100m × 0.25mm, ID) and RTX-5(1.2m × 0.18mm, ID) respectively; primaryoven temperature program of 150℃ for 1min, 150-220℃ at 3℃/min and 220℃ for 20min, also secondary
oven temperature was increase 20℃ of primary oven temperature; Modulator temperature offset was 30℃;
second dimension separation time was 6 sec; hot pulse time 1.20 sec and cool time between stages was 1.80 sec.
The structural composition of TFAs in the triacylglycerols of the oil extracted from commercial butter andmargarine has been studied using NMR spectroscopy. The intensity of the signal can also be used for quantitation.
In the case of fat from butter and margarine, C18 carbon allylic to a trans bond has a chemical shift of 32.6 ppm.
Key word: Trans fat, GCxGC, NMR
Introduction
In recent years there has been an increasing awareness of the influence of diet on human well-being,
especially the effect of dietary fat on heart disease. It has been studied that partially hydrogenated vegetable oils
contain trans unsaturation, positional isomers and conjugation not naturally occurring in vegetable oils. Although
the major emphasis has been on the fatty acid composition of food fats, other characteristics have been
implicated. There include triglyceride structure and configurational and positional isomers. Since the effect oftrans fats on the human health may be different depending the positional isomer, the separation of individual
isomer of trans fats (TFAs) has been attracted a great interest by the many separation scientists. TFAs are present
in meat and dairy products as a by-product of fermentation in ruminant animals as a nature and in vegetable fats
as a result of partial hydrogenation. This study was aimed to discrimination of natural(N-TFA) and hydrogenatedtrans fatty acid(H-TFA) by comprehensive GC×GC and NMR spectroscopy.
Materials and Methods
Lipids from butter and margarine for the TFA analysis were extracted by chloroform and methanol mixture(2:1, v/v) and evaporated to dryness. Methyl ester of TFA was obtained by the reaction in a heating block with
BF3/Methanol as a catalyst. For the NMR analysis of TFA, 50 mg of lipid was taken and solved in 500 uL of
CDCl3.
GCxGC analysis was performed using LECO system. The column set for GCxGC analysis consisted of twocolumns. The primary column was SP-2560(100m x 250um x 0.2um, film thickness) and secondary column was
RTX-5(1.5 m x 0.18mm x 0.18um, film thickness).
NMR analysis was performed using Bruker Avance III 400 (Bruker, Germany). 1H-NMR (500 MHz),13
C-NMR (125 MHz), 1D and DEPT, were used to predict the TFA content in butter and margarine, and estimate
the position of double bond.
Corresponding author, Jaeho HA,516, Baekhyeon, Bundang, Seongnam, Gyeonggi, 463-746, R.O.KOREA,email :jhkfri@kfri.re.kr.

Leader of Food Analysis Division of Korea Society of Food Science and Technology
Vice President of Korea Society of Analytical Science
Visiting Professor/Tsinghua University, China
Director of Food Analysis Center and Food Safety Research Division
Delegate of CODEX for the Analytical Method in Korea

Result and Discussion
The key isomers of TFA to discriminate the natural (N-TFA) and hydrogenated (H-TFA) TFAs are w9 andw11 TFAs since these two isomers are the major TFA with one double bond. To discriminate and quantify theamount of these two TFAs, the optimum analytical condition has been studied. For the separation of these two
isomers, The condition of comprehensive GC×GC has been optimized: primary oven temperature program of
150℃ for 1min, 150-220℃ at 3℃/min and 220℃ for 20min, also secondary oven temperature was increase 20℃
of primary oven temperature; Modulator temperature offset was 30℃; second dimension separation time was 6
sec; hot pulse time 1.20 sec and cool time between stages was 1.80 sec. Since w9 and w11 TFA isomers, which
are most abundant in the lipids in both natural and hydrogenated TFAs, were well separated.
High resolution 13C NMR spectroscopy has several applications in fat analysis and is an effective tool thatcan provide structural information about cis and trans isomers of fatty acid. The signal from a carbon that is
allylic to a double-bond carbon provides the evidence concerning the cis and trans nature of the double bond. The
intensity of the signal can also be used for quantitation. In the case of fat from butter and margarine, C18 carbon
allylic to a trans bond has a chemical shift of 32.6 ppm.
References
1.F. Destaillats et al., Comparison of available analytical methods to measure trans octadecenoic acid isomericprofile and content by gas liquid chromatography in milk fat, J. Chromatogr. A 1145 (2007) 222-228
2.M. Adahchour et al., Recent developments in the application of comprehensive two-dimensional gas
chromatography, J. Chromatogr. A 1186 (2008) 67-108
3.J.M. Chardigny et al., Rationale and design of the TRANSFACT project phase 1: A study to assess the effect of
the two different dietary sources of trans fatty acids on cardiovascular risk factor in humans, Contemporary
Clinical Trials 27 (2006) 364-373
4. Casimir C. Akoh and David B. Min, Food Lipids, Marcel Dekker Inc., New York, NY 840, pp.175=176, 1998.

Corresponding author, Jaeho HA,516, Baekhyeon, Bundang, Seongnam, Gyeonggi, 463-746, R.O.KOREA,email :jhkfri@kfri.re.kr.

Leader of Food Analysis Division of Korea Society of Food Science and Technology
Vice President of Korea Society of Analytical Science
Visiting Professor/Tsinghua University, China
Director of Food Analysis Center and Food Safety Research Division
Delegate of CODEX for the Analytical Method in Korea

Monitoring of Hazard Air Pollutants (HAPs) Inventory
Gon Ok1),2), Jee-hoon Kim1), No-jin Park1),2), Seok-hyung Lee1),2), Sung-min Hwang1),2), Dong-hwan Kim1)
1) Graduate School of Earth Environmental Engineering, Pukyong National University
2) Dioxin Research Center, Pukyong National University

Summary
The Hazard Air Pollutants (HAPs) not only risk human health by the direct inhaling but also
leads to the pollutions in different environmental medias, such as plants, water and soil
through atmospheric pathways. In Korea, 53 chemicals are selected as Hazard Air Pollutants.
However, there is not enough data on the source and the amount of HAPs that are being
produced. This study was the research at national level on producing HAPs source inventory
and the emission estimation method. Additionally, the methods on analyzing HAPs from thevarious sources were organized. Also, the field monitoring on HAPs emission from synthetic
resin and plastic manufacture industries, one of the priority sources of HAPs. According to theresult, the amount HAPs emission in this study was lower than the level reported in TRI.
However, more various types of compounds were detected in this study than the compounds
reported by TRI.
Key word: Hazard Air Pollutants (HAPs), Sources, Inventory, GC/MS
1. Introduction

The hazardous compounds take various pathways to enter human body. Humans inhale 15kg
of air daily, which is 10 times more than the intake of water and food. The human exposure to
HAPs through inhalation is crucial because the substances are directly absorbed into lung andhuman body. Furthermore, HAPs that are released in ambient air have high potency ofharming ecosystem, as it is deposited in plants, soil and water. Since HAPs are toxic
compounds that directly and indirectly pose negative influences on human health, plants andanimals, different countries selected certain chemicals as HAPs. For example, United States,
Germany, Japan and Korea selected 187,154,234 and 53 compounds as HAPs, respectively.
In Korea, there are Toxic Release Inventory (TRI) and Stack Emission Management System(SEMS) as HAPs data. However, TRI and SEMS have certain restrictions that prevent moreaccurate data production. In case of TRI, the concentration levels are the values reported by
the industries that produced HAPs, while SEMS only involves the level of certain
compounds released from the chimney. In this study, the attempt for the creation of moreaccurate and realistic inventory at the national level was made.Furthermore, the source
monitoring of HAPs was held based on the suggested list of different sources of HAPs, andthe results were compared with the already existing data.
2. Materials and Methods
2.1 HAPs inventory
Through TRI, SEMS and other reference surveys, the main industries that are sources of
HAPs production were selected and listed. The concentration estimation method wassuggested using Top down approach, and the emission factor and activity research method forthe main HAPs sources were suggested.
2.2 SamplingSample collection
The source of HAPs can be classified into two groups, point sources and non-point sources.
For the point source analysis, the analytical methods suggested by Korean standard method
and U.S EPA methods, depending on the chemical compound group. Non-point sources that
may leak, such as flange, valve and pump have various types and appearances, and there does
not exist sampling and monitoring method specific to those non-point sources. Thus, twomonitoring methods for non-point sources were proposed after the numerous testing, and wereapplied in actual sampling and analysis.
- Vacuum Bagging method
The leakage point at the valve was covered with Teflon film and tightly sealed on the pipe,
creating a bag. The air inside the bag was pumped out. As the gas leakage occurs from valve,
the bag expands, and the glass filter is exerted to collect the leaked gas. However, the vacuum
bagging method is only possible when the amount of leaking gas is noticeably a lot, and when
there is no danger of explosion. Thus, the vacuum bagging method is inappropriate for theusage at petrochemical industry plants.
- Blow-through bagging method
The leakage point at the valve was covered with the film and tightly sealed on the pipe tocreate a bag. At the entrance of the sealed bag, the high purity nitrogen was injected, and thegas released from the opposite opening was collected. During the process of collecting gas
samples in both vacuum bagging method and blow through bagging method, five different
strategies, Tedlar bag, Absorption tube, 2,4-DNPH cartridge, Filter & Adsorbent & Impinger,
Impinger, absorption tube, were applied depending on the target compound.
Instrumental Analysis
For the analysis of five different target compounds, 6 types of instrumental analysis methods
were used. For the analysis of VVOC groups, Pre-concentrator and GC/MS (Entech andAgilent) were used. Thermo Desorption System and GC/MS (Gerstel TDS and Agilent) wereused for the VOC groups, while Agilent GC.MSD was used for the PAHs and Aldehyde group
analysis. Finally, the HCl was analyzed by UV-vis.
2.3 Monitoring
The monitoring for the HAPs sources were held in 5 plastic manufacture plants that produces
HDPE, LHDPE, LLDPE,PP,PE,PVC and ABS. In possible source points in each plant, 5samples were collected from non-point sources, 2 point sources and 2 background samples.
3. Result and Discussion
HAPs emission inventory had 77 priority sources form 12 main categories of original CAPSS.
The field monitoring on HAPs emission from synthetic resin and plastic manufactureindustries, one of the priority sources of HAPs. According to the result of comparison of TRI
and the field monitoring of 53 HAPs compounds, the field monitoring showed more various
types of HAPs compounds. The TRI reported 10 HAPs compound, while the field monitoringin this study reported 14 HAPs compounds out of 53 compounds. However, the amount of 53
HAPs emission was less than the emission amount reported by TRI. Such results wereconfirmed to be caused by the application of TRI default value, which led to theoverestimation than the actual field monitoring emission amount.

Biography : Professor Gon Ok. Ph.D
Professor of Department of Environmental Atmospheric Science andGraduate School of Environmental Atmospheric Science,
Chief Professor of Graduate School of Earth Environmental Engineering,
And the Director of Dioxin Research Center
Pukyong National University.
Address: 599-1 Daeyeon 3-dong, Nam-gu, Busan, 608-737, KOREA.
Tel : +82-51-629-6641
Fax : +82-51-626-9635
E-mail : gonok@pknu.ac.kr

Dr. Gon Ok is a professor at Department of Environmental Atmospheric Science, Graduate School ofEnvironmental Atmospheric Science and the Chief Professor of Graduate School of Earth Environmental
Engineering at Pukyong National University. Professor Gon Ok has been working as a professor at Pukyong
National University since 1995. Professor Gon Ok is the Director of Dioxin Research Center at Pukyong
National University in Busan, Korea.
Prof. Gon Ok received his Ph.D. in Materials and Environmental Safety Engineering at Yokohama National
University in Japan.
He is currently an academic chair and the editorial board member of the Korean Society for Environmental
Analysis.
Professor Gon Ok was the chief researcher, and the member of Committee Board, for the development of
Dioxins (PCDD/DFs and DLPCBs, 1995) and POPs(2005) Standard Method research, conducted by theenvironmental ministry of Korea.
He carried out the research for the development of Waste Treatment Technology and Management Guideline
in 1996. Furthermore, from 2001 to 2005, he developed the National Inventory Method and Dioxins (PCDD/DFs,
PCBs,HCB) monitoring methods in Korea.

Professor Gon Ok was the chief researcher of Dioxins Exposure and Health Assessment study organized by
the Korean Ministry of Environment in 2005 and 2006. From 2002 to 2008, he also led the research on
Dioxins, Endocrine Disruptors, and POPs Assessment in Marine Eco-Environment, organized by Korean
Ministry of Maritime Affairs & Fisheries.
In 2006, Professor Gon Ok was the Water, Land and Nature Steering Committee member of the Presidentialcommission on Sustainable Development in Korea.
Professor Gon Ok is currently conducting researches on the Development of National Inventory of HAPs
(Hazards Air Pollutants) by Ministry of the Environment Republic of Korea.
He also works for the development of monitoring method and Inventory, environmental monitoring inmedia, path- way and long range transportation monitoring, and the assessment on emerging POPs(PBDEs,
PCNS, HCHs and PFCs such a PFOS/PFOA et.) in Asian Dust.

On-Line and On-Column Electrochemical Redox Derivatization
Enhancement of Separation Selectivity of Liquid ChromatographyforM. Shibukawa1
1

Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570, Japan

Summary
On-line and on-column electrochemical redox derivatization methods for enhancement of separation selectivity of
HPLC are presented. In the former separation system, the derivatization reaction proceeds in a small flow through electrolytic
cell placed between two separation columns so that an analyte compound migrates as its original form in the first column,
while as its converted form in the second column. On the other hand, in the latter strategy an LC column designed forelectrochemically modulated liquid chromatography (EMLC) is used for derivatization as well as separation. We can gain
broad and facile control by these techniques over the distribution of redox species in the separation system and achieve highly
selective separation.
Key word: Electrochemical redox derivatization; Electrolytic flow cell; Electrochemically modulated liquid chromatography;
Secondary chemical equilibrium

1 Introduction
HPLC is always demanded to acquire new separation selectivity in order to achieve better separation anddetermination of trace levels of compounds in complex matrix samples. Use of a chemical reaction specific for target
compounds as a pre-column or on-column derivatization is one of the efficient ways for enhancement of separation selectivityof HPLC. We recently presented two redox derivatization techniques, on-line derivatization using double separation columns
and one redox derivatization unit [1, 2] and on-column derivatization via electrochemically modulated liquid chromatography(EMLC) [3]. It will be shown in this paper that on-line and on-column electrochemical redox derivatization techniques provide
intriguing pathways for realizing a highly selective separation of redox active analytes.

2 Materials and Methods
The electrolytic flow cell used for the on-line electrochemical derivatization was a Model 5020 guard cell (ESA,
USA), which contained a porous graphite working electrode, a palladium counter electrode and a palladium-hydrogenreference electrode (Pd/H2). All potentials described for this cell are given vs. Pd/H2.
The EMLC column consists of a Nafion cation-exchange membrane in tubular form (Perma Pure Inc., USA) placed
inside a porous stainless steel cylinder (Mott Metallurgical Corp., USA) [4]. The Nafion tubing serves as a container for the
porous graphitic carbon (PGC) stationary phase (3.5 mm BTR carbon, Biotech Research, Japan). PGC also functions as theworking electrode. The porous stainless steel cylinder prevents deformation of the Nafion tubing under the high pressure ofchromatographic flow and is also used as the counter electrode. An Ag/AgCl reference electrode (saturated NaCl) is placed inan electrolyte reservoir that surrounds the stainless steel counter electrode.

3 Results and Discussion
The on-line electrochemical redox derivatization HPLC system consists of two separation columns and a small
electrolytic flow cell placed between them [2]. A redox derivatization reaction proceeds in the electrolytic flow cell so that an
analyte compound migrates as its original form in the first column, while as its oxidized or reduced form in the second column.

The selective separation of an analyte compound can be achieved because redox reactions specific for the analyte in theelectrochemical derivatization unit can be controlled by alteration of potential applied to the cell. There are three requisites

for the cell to be used for the on-line redox derivatization HPLC: (1)the cell should withstand high pressure; (2)the innervolume of the cell should be small; and (3)the efficiency of electrolysis should be 100 %. We substantiated that theelectrolytic cell we adopted meets the first two requisites; the maximum operating pressure of the cell shown by the
manufacturer is 422 kgf/cm2 and the inner volume of the cell we determined is 44 mL. We also evaluated the electrolysis
efficiency of the cell using Co(II)-EDTA and Co(III)-EDTA as model compounds and found that Co(II)-EDTA wasoxidized to Co(III)-EDTA by the electrolytic cell at 0.1 V or above, while Co(III)-EDTA was completely converted to theCo(II)-EDTA at -0.6 V or below. The electrolysis efficiency is approximately constant independent of the flow rate in therange of 0.02 to 0.10 mL min-1. We applied this on-line electrochemical redox derivatization system to determination of
trace amount of cobalt in a stainless steel sample after chelation with EDTA. The cobalt complex eluted in the firstcolumn as Co(II)-EDTA and was separated from the trivalent metal complexes including Fe(III), while after oxidation by
the electrolytic cell it was completely separated from the other divalent metal ions during elution in the second column.
The cobalt concentration determined by the present method was in good agreement with the recommended value.
Using catalytic redox activity of PGC packings, we presented a new type of secondary chemical equilibrium(SCE) for enhancement of selectivity in HPLC [5]. This technique is based on redox equilibria and may become much
more useful if the equilibria can be controlled electrochemically. We have thus developed an on-column electrochemical
redox derivatization method using EMLC technique [3]. We examined dependence of the retention factor and peak areaof hydroquinone, resorcinol, catechol, tyrosine, and dopa on potential applied to the EMLC column and found out thathydroquinone and catechol migrate through the column as equilibrium mixtures of their oxidized and reduced forms andthe heterogeneous rates for these redox reactions are much higher than the rate of mass transfer between the mobile andstationary phases. This indicates that these redox reactions can be used as secondary chemical equilibria so that thecorresponding equilibrium mixtures elute as single peaks and their retention times can be controlled by alterations in thepotential applied to the PGC stationary phase. All five compounds were reasonably separated at +300 mV, whereas both
catechol and resorcinol and dopa and L-tyrosine co-eluted at 0 mV. These data point to the ability to enhance
EMLC-based separations via on-column electrochemical transformations of the analyte. Factors that influence efficiency
in EMLC will also be reviewed from a chromatographic perspective.
4 References[1] K. Saitoh, N. Yamada, E. Ishikawa, H. Nakajima, M. Shibukawa, J. Sep. Sci. 2006, 29, 49.
[2] K. Saitoh, S. Naitoh, M. Endo, M. Washiya, M. Shibukawa, Bull. Chem. Soc. Jpn. 2007, 80, 951.
[3] Kazunori Saitoh, K. Koichi, F. Yabiku, Y. Noda, M. D. Porter, M. Shibukawa, J. Chromatogr. A 2008, 1180, 66.
[4] E. Y. Ting, M. D. Porter, Anal. Chem. 1998, 70, 94.
[5] M. Shibukawa, A. Unno, T. Miura, A. Nagoya, K. Oguma, Anal. Chem. 2003, 75, 2775.

5 Biographical Sketch
Name : Masami Shibukawa
Nationality : Japanese
Affiliation : Graduate School of Science and Engineering, Saitama University
Contact : E-mail: sibukawa@apc.saitama-u.ac.jp
Web: http://www.apc.saitama-u.ac.jp/bunseki/Index1.html
Title : Professor

Preparation, Characterization, and Analytical Applications of a Novel Hybrid
Organic-inorganic Material Used as Chromatographic Stationary Phase
Yan Zhu* and Yingying Zhong
Department of Chemistry, Xixi Campus, Zhejiang University, Hangzhou 310028, Zhejiang, P. R. China
Email:zhuyan@zju.edu.cn
So far, columns of packed particles have still been the most popular and core devices forchromatographic separations and detections because of their great utility, excellent performance and wide
variety. In order to overcome the defects of traditional column packing, such as fragile under extreme pH
and not rigid enough under high pressure, a novel hybrid organic-inorganic material used as
chromatographic stationary phase has been synthesized, which maintains the advantages of both organic
polymers and inorganic supporters.
According to the excellent mechanical and thermal properties, carbon nanotubes with a concentration
between 0.1~5% (wt%) of monomers have been incorporated into polystyrene-divinyl benzene
microspheres to form homogeneous packed particles ranging from 2 μm to 8 μm. Three different ways havebeen tried to synthesize polystyrene-divinyl benzene-carbon nanotubes (PS-DVB-CNT) hybrid particles.
One is to intermingle carbon nanotubes which is purified by mixed acids with monomers simply, another is
to copolymerize modified carbon nanotubes with other monomers, and the third is to modify the surface ofpolystyrene-divinyl benzene with pretreated carbon nanotubes. After optimization of the synthetic
conditions, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM),
thermal-gravimetric analysis (TGA), elemental analysis and Fourier transform Laman spectroscopy(FT-Laman) have all been used for the characterization of this novel stationary phase. And the results provethat carbon nanotubes have been incorporated into the matrix of polystyrene-divinyl benzene definitely.
Besides, the chromatographic properties of the stationary phase have been investigated byreversed-phase high-performance liquid chromatography with alkylbenzenes, pyridines, anilines andphenols as probes. After comparison with pure PS-DVB particles, the hybrid stationary phase owns betterhydrophilicity and the chromatographic peaks are more symmetric, which may help to separate non-polar
compounds on organic polymer supporters. And with wide pH endurance, this stationary phase can beutilized in extreme mobile phases to separate different kinds of acids and alkalis. Meanwhile, it is suitable
to be used in ion-pair chromatography to separate organic and inorganic acids for its longer life comparedwith silica gel columns. As a result, the incorporation of carbon nanotubes has enhanced the rigidity of thepacked particles and improved the porous structure of the microspheres with better selectivity, resolution
and wider applications.

Biography
Professor Dr. Yan Zhu
Department of Chemistry, Zhejiang University, Hangzhou 310028, China
Tel/Fax: +86-571-88273637
E-mail: zhuyan@zju.edu.cn

Professor Yan Zhu was born in 1963 in Hangzhou, China. He received his BA degree in 1986
at Hangzhou University and received a PhD degree in environment science at Zhejiang
University in 2001. He had studied and worked in University of Tasmania, Australia during
2001-2002. He is currently a full professor of Zhejiang University since 2008. He is amember of council of Chinese Society for Chromatography Science and a member of councilof Analytical Instrument Society, Chinese Instrument Society. He is a contributing editorial
broads Chinese Journal of Chromatography (Sepu) and several Chinese Journals. He received
several awards for his contributions in ion chromatography and separation science:
Outstanding Young Scientist Award of Zhejiang Provice (Zhejiang Government, 1996),
Achievement Award for Ion Chromatography (Ion Chromatography Discussion Group, Japan
Society of Analytical Chemistry, 2006) , Science Fund for Distinguished Young Scholars ofZhejiang Province (Natural Science Foundation of Zhejiang Province, 2008).
His current research is focused on ion chromatography, new stationary phase for liquid
chromatography, and sample pre-treatment for ionized species. He is the author and co-authorof 150 original research papers published in international journals, 5 reviews, 4 books and 10
patents.

Use of ion-exclusion/adsorption chromatography to evaluate quality of TiO2
photocatalytic materials in water
Masanobu Mori 1, Kazuhiko Tanaka 2, Hiroshi Taoda 3, Hideyuki Itabashi 1
1Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan

2

Graduate School for International Development and Cooperation, Hiroshima University, Japan

3

National Institute of Advanced Industrial Science and Technology, Japan

Summary
In our laboratory, ion-exclusion/adsorption chromatography of dimethylsulfoxide (DMSO) and its derivatives, i.e.,methanesulfinic acid (MSI), methanesulfonic acid (MSA), and sulfuric acid (SA), was employed to test the quality of TiO2
photocatalyst in water. DMSO was separated by hydrophobic adsorption, and MSI, MSA, and SA were separated byion-exclusion using a weakly acidic cation-exchange resin column with 20 mM succinic acid as the eluent. Among theseparated species, DMSO and MSI were detected by UV at 195 nm and MSA and SA were detected by conductivity. Thismethod was used to monitor the decomposition of DMSO with OH radicals generated by the UV irradiation of TiO2-coatedmaterials. The concentration of DMSO by OH radicals was found to decrease through the stoichiometric reaction in the orderof MSI, MSA, and SA.
Keywords: Ion-exclusion; adsorption; chromatography; photocatalyst; dimethylsulfoxide; OH radicals
1 Introduction
Recently, the use of TiO2 photocatalyst has attracted from the viewpoint of applications in advanced oxidation technologies
for the purification of air and water. Therefore, many TiO2-coated materials have been developed by researchers, and they havebeen employed commercially in the manufacture of tiles, floors, walls, roads, glass, etc.
In order to establish a quality test for photocatalytic materials in the liquid phase, we used dimethylsulfoxide (DMSO) as atest sample. DMSO is well known as a scavenger that can be used to indirectly determine active oxygens such as OH radicals
generated during a biological reaction [1]. By the oxidative decomposition of DMSO with OH radicals, MSI, MSA, and SA
are formed on the basis of stoichiometric reaction. In this study, we developed an ion-exclusion/adsorption chromatography of
DMSO and its derivatives as an analytical method for evaluating the artificial decomposition of DMSO by a photocatalytic
material.

2 Materials and Methods
Ion-exclusion chromatographic system
We used the Tosoh IC-2001 ion chromatograph; it consists of a dual pump (0.6 ml/min), an online degasser, an auto-sample
injector (30 μl), a column oven (40 °C), and a conductivity detector, and the Tosoh UV 8020 UV-vis detector was inserted
between the separation column and the conductivity detector. We used the Tosoh TSKgel Super IC-A/C as a separation
column; this is a polymethacrylate-based weakly acidic cation-exchange resin of H+-form that is packed with 3-μm particles,
and it has 0.1 mequiv./ml cation-exchange capacity (150 mm × 6 mm i.d.). Among the species separated by the column,
DMSO and MSI were detected by UV at 195 nm and MSA and SA were detected conductivity.
Masanobu Mori, m_mori@chem-bio.gunma-u.ac.jp

Photocatalytic reaction system
A schematic of the acryl-resin-based photocatalytic oxidation reactor (290 × 108 × 110 mm) used in this system is shown in
Fig. 1. The tested TiO2-coated materials were placed in the photocatalyst area of the reactor. A 500-ml solution containing 10
ppm DMSO was poured into the photocatalytic reactor. An anatase-type TiO2-glass beads BL2.5DX coating 1 μm thickness of
TiO2 on a 2.5mm Ø of silica-glass bead was purchased from Photocatalytic Materials Inc. (Aichi, Japan). 50 g ofphotocatalyst-coated beads were used in one experiment. The
TiO2photocatalyst was a commercial BLB light (λMAX = 350 nm).

Lamp

The solution in the reactor was circulated by a pump at a flowrate of 500 ml/min. During the photocatalytic reaction, a 10-ml

Test solution

sample solution was collected every 30 min. The concentrationsof DMSO and its products in the solution were determined by
the ion-exclusion/adsorption chromatography techniquedeveloped in this study.

Turbulent piece

Photocatalyst

Fig. 1. Schematic illustration of the present
photocatalytic reaction system

3 Result and Discussion
Ion-exclusion/adsorption chromatography of DMSO and its derivatives using a weakly acidic cation-exchange resin in the
H+-form with succinic acid as the eluent was optimized in terms of the retention times and the peak resolutions of each analytewith UV and conductivity detections. In ion-exclusion/adsorption chromatography, the retention mechanism is hydrophobic
adsorption onto the cation-exchange resin for organic solvents such as DMSO [2]; in contrast, the mechanism is ion-exclusion
for fully ionized species such as MSI, MSA, and SA [3]. As a result, a well-resolved chromatogram was obtained with the
combination of a weakly acidic cation-exchange resin column and 20 mM succinic acid.
The photooxidation of DMSO by anatase-type TiO2-coated glass beads in water was investigated by using the photocatalytic
reaction system developed in this study. The test solution with 10 ppm DMSO was poured into the reactor and circulated for 4
h under UV irradiation. As shown in Fig. 2, the remaining DMSO decreased gradually with the time-dependence of UV
irradiation; in contrast, the intermediates were produced in the order of MSI, MSA, and SA with irradiation. As shown in Fig.
3, the plots indicated that the concentrations of sulfur atoms in the remaining DMSO and the intermediate products formed
were found to be stoichiometric oxidation by OH radicals from the TiO2 material irradiated with UV light.
In Japan, one part of this analytical method was adopted as a standardized evaluation method for TiO2 photocatalytic
materials in water (JIS R1704), and at present, it is awaiting ISO certification (ISO/CD10676 Fine ceramics - advanced
ceramics, advanced technical ceramics).

4 References[1] R. Bruck, H. Aeed, H. Shirin, Z. Matas, L. Zaidel, Y. Avni, Z. Halpern, J. Hepatol. 31 (1999) 27.
[2] M. Mori, K. Tanaka, M.I.H. Helaleh, Q. Xu, M. Ikedo, Y. Ogura, S. Sato, J. Chromatogr. A, 997 (2003) 219.
[3] K. Tanaka, J.S. Fritz, J. Chromatogr. 409 (1987) 271.

Masanobu Mori, m_mori@chem-bio.gunma-u.ac.jp

DMSO

1h

2h

Abs. at 195 nm

0h

4h

MSI

Conductivity (μS/cm)

Time (min)MSA
SATime (min)Fig. 2. Peak profiles of decomposed DMSO and the products formed by the TiO2 photocatalyst with UV irradiations.
Ion chromatographic and photocatalytic conditions are described in Materials and Methods (i.e., Sec. 2).
Non-irradiation

Concentration of sulfur (ppm)10

Total-S

8

DMSO

MSA

6
4

MSI

2

H2 SO4
0
0

60

120

180

240

Fig. 3. Changes of sulfur atom concentrations in
the DMSO and its decomposed products with andwithout UV-irradiation

Irradiation time (min)5 Biographical Sketch
Name: Masanobu MORI
Born: Hokkaido, Japan (01/07/1971)
Nationality: Japanese
Affiliation: Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan
Contact: E-mail: m_mori@chem-bio.gunma-u.ac.jp
Web: http://www.chem.gunma-u.ac.jp/~md3/
Title: PhD (Environmental Science, Hokkaido University, 2001)
Record:2001–2005

National Institute of Advanced Industrial Science and Technology, Post Doctoral Researcher

2004–2005

Photocatalytic Materials Inc., Researcher

2005–Present

Graduate School of Engineering, Gunma University, Associate Professor

2007–2008

University of Texas at Arlington, Visiting Professor

Masanobu Mori, m_mori@chem-bio.gunma-u.ac.jp

Analysis of Amino Acid and Peptide Biomarkers by CEC-ESI-MS andHPLC-ECD
Zilin Chen*, Yue Wu, Lihong Liu, Fang Wang
Department of Pharmaceutical Analysis, College of Pharmacy,
Wuhan University, Wuhan, 430072, China
chenzl@whu.edu.cn

Some amino acids and peptides are regarded as biomarkers which areassociated with human diseases. For examples, amyloid-β(Aβ)polypeptides istargeted as the potential biomarker for Alzheimer’s disease (AD) [1]. The oxidized
tyrosine derivatives including 3,4-dihydroxytyroisine (L-DOPA), dopamine,
dopamine quinine are relevant in Parkinson’s disease [2]. Therefore, the analyticaltechniques for analysis of multiple amino acids and peptides are needed as a newparadigm in disease prognosis and drug development. Hyphenated separation
techniques like liquid chromatography (LC), capillary electrophoresis (CE) andelectrochraomatography (CEC) coupled to mass spectrometric (MS) andelectrochemical detections (ECD) offer high separation selectivity and excellent
detection sensitivity.
In recent years we focus one of our works on the development of hyphenatedseparation techniques CE/CEC/LC with MS and ECD detections [3-5]. The recent
progress on analysis of amino acid and peptides by CEC-ESI-MS and LC-ECD in mygroup will be presented in this talk. The main contents include (1) a novel sheathlessnanoelectrospray interface for hyphenating CE/CEC to electrospray (ESI)-MS [3], (2)Lupamin-coated column technology for analysis of amino acids and peptides by
CEC-ESI-MS [3,4], (3) analysis of small molecular peptides with same amino acidcomposition but different sequence by CE and LC-MS [5], and (4) analysis of aminoacid biomarkers of tryptophan and kynurenine by LC-ECD with carbon
nanotubes-modified electrodes.
References[1] N. J. Clarke*, F. W. Crow, S. Younkin, S. Naylor, Anal. Biochem. 298 (2001) 32-39.
[2] A. S. Ptolemy, R. Lee, P. Britz-McKibbin*, Amino Acids 33 (2007) 3-18.

[3] Z. Chen*, B. Boggess, H. C. Chang, J. Mass Spec. 42(2007)244-253.
[4] Y. Wu, J. Xie, F. Wang, Z. Chen*, J. Sep. Sci. 31 (2008) 814.
[5] Y. Wu, J. Xie, F. Wang, Z. Chen*, J. Sep. Sci. 32(2009)437-440.
Acknowledgments
This work was supported by the National Scientific Foundation of China (NSFC NO.20775055
and NO. 90817103), Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry and the start-up funding for Luojia Chair Professorship of Wuhan
University (No. 306276216).

Biography
Dr. Zilin Chen
Luojia Scholar-Chair Professor, Department Chair
Department of Pharmaceutical Analysis and Screening
College of Pharmacy
Wuhan University
Wuhan, 430072, China
Email: chenzl@whu.edu.cn
Phone: 86-27-68759893
Fax: 86-27-68759850

Dr. Zilin Chen received his Ph D degree at Tokyo Metropolitan University (TMU),
Japan, in March, 2000 and then worked as a postdoctoral fellow of JSPS (Japan
Society for Promotion of Science) at TMU for 2 years with Prof. Dr. Toshiyuki Hobo.
After having worked at NTT Science and Core Technology Laboratory Group for
another 2 years, he moved to University of Notre Dame, USA in 2004 and worked asa research assistant professor.
Dr Chen joined Wuhan University in March, 2007. He is a Luojia Scholar-Chair
Professor of Wuhan University and department chair of Pharmaceutical Analysis andDrug Screening. He serves as editorial board members for international journal “Drug
Discoveries & Therapeutics” and national journal “Herald of Medicine”. He wasawarded “New Century Prize” by Japan Society for Analytical Chemistry (JSAC) and“Encouragement (Shourei) Award” by Japan Society for Chromatographic Sciences(JSCS) in 2003. He has published over 60 peer-refereed journal articles. Dr. Chen’s
research interest is micro-scaled separation and detection for bio- and/or
pharmaceutical samples.

Advance of capillary electrophoresis/electrochromatography coupling withelectrospray ionization mass spectrometry based on monolithic columns in
doping analysis
Minghua Lu, Xin Li, Qiaomei Lu, Guonan Chen, Lan Zhang*
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety(Fuzhou University), Department of Chemistry, Fuzhou University, Fuzhou 350002, Fujian, China

zlan@fzu.edu.cn
In recent 30 years, capillary electrophoresis/electrochromatography (CE/CEC)
have attracted considerable attention due to high separation efficiency, low sample
consumption and short analysis time. Compared with ultraviolet detector, which usedmostly in CE/CEC instruments, mass spectrometry (MS) has some inherent
advantages. Firstly, MS can provide both molecular mass and structural information.
Secondly, MS enables to extract analytes signals from a high background noise.
Thirdly, MS can provide high selectivity and sensitivity. These are of particularimportance when it is necessary to positively identify analytes in complex samples
such as biological fluids.
Monolithic column technique is an emerging column preparation technique, andattracted more and more attention due to good permeability, fast mass transfer
property, high stability and easy modification. Monolithic columns can be classified
into two categories: silica-based and organic polymer-based monoliths. Silica-basedmonoliths usually offer the advantages of high mechanical strength, heat stability andresistance to organic solvent. In comparison with silica-based beds, polymer-basedmonoliths exhibit more potential advantages, involving simple preparation and thepossibility of controlling their properties of porous and surface chemistries.
In our laboratory, some novel CE/CEC-ESI-MS methods based on different kinds ofcolumns, which including bare fused capillary, chemically modified fused silica
capillary, silica-based and polymer-based monolithic column, have been developed
for rapid and sensitive analysis of doping. A rapid, high-resolution and effective new
method for simultaneously analyzing 12 diuretics by CE-ESI-MS was demonstrated in
our laboratory [1]. To reduce adsorption and provide fast separation ability, a coatedcapillary was prepared and applied to rapid analysis of peptides and amino acids in
CE-ESI-MS system [2]. Based on silica-based monolithic columns, apressure-assisted CEC-MS method for rapid and sensitive analysis of narcotics wasestablished. Under the optimum conditions, the tested five narcotics could becompletely separated within 10 min with the detection limit in the range of 2.0–80
nmol/L [3]. A method for rapid separation and sensitive detection of β-blockers bypressure-assisted CEC-ESI-MS using silica-based monolithic column were within 11
min with detection limits in the range of 0.15–0.80 ng/mL (deﬁned as S/N= 3). The real
samples from some male volunteers were successfully analyzed and confirmed withthe proposed method [4]. Also, to overcome limitation of silica-based monolithic
columns, a novel porous polymer-based monolithic column was prepared [5] andapplied to analysis of β2-agonist and narcotics [6]. These novel CE/CEC-ESI-MS
methods present many superiority compared with traditional GC-MS or LC-MS
methods, which include higher separation efficiency, shorter analytical time, and lower
sample and organic solvent consumption.
Referents:[1] Electrophoresis 2007, 28, 1461-1471. [2] Electrophoresis 2009, 30, ***-*** (available online)[3] Electrophoresis 2008, 29, 936-943. [4] J. Chromatogr. A, 2008, 1193, 156-163. [5]Electrophoresis 2009, 30, ***-*** (accepted) [6] In preparing.

Dr. and Prof. Lan Zhang,Prof. Lan Zhang was born in March 1963. She received her M.S.
PhD degree in Analytical Chemistry from East China Normal University.
She has been a teacher of Fuzhou University in 1995 and now she ishead of the Fuzhou University Detect Center. Her research interests
cover the pharmaceutical analysis, bioelectrochemistry, capillaryelectrophoresis, Choromatographic Technology, Chromatography andmass spectrometry combined technology, etc. Prof. Lan Zhang has
gained the brilliant academic achievements in pharmaceuticalanalysis, modern separation and detection technology, etc. More than
70 papers have been published in the international Journals such as
Electrophoresis, J. Chromatogr. A, Talanta, Anal. Chem. Acta , etc.
and gained much citation. In 2002, Prof. Lan Zhang was elected for
the Fujian Provincial Project of “Talent Project”. In 2006, Prof. Lan
Zhang was inducted into “the Plan of University New Century
Outstanding Talented Man of Fujian Province”. Then Prof. Lan Zhang
received the Second Prize of Fujian Provincial Scientific-Development
Award in 2004 and 2006 respectively. Now Prof. Lan Zhang continue
her scientific career as the Director of University Analytical andTesting center of Fujian Province and a main member of the Ministry of
Education Key Laboratory of Analysis and Detection for Food Safety.
She immersed in the research of the key technology for food safety
capillary electrophoresis and capillary electrochromatography,
chromatography and mass spectrometry combined technology, theseparation and analysis of natural medicinal effective compounds andbioactive compounds, etc.
Representative paper：
1. Minghua Lu, Lan Zhang*, etc. “Rapid analysis of peptides and amino acidsby

CE-ESI-MS

usingchemically

modiﬁedfused-silica

capillaries”,

Electrophoresis 2009, 30, 1-7.
2. Wei Liu, Lan Zhang*, etc. “Analysis of agonists and blockers in urine using
hollow fibre - protected liquid-phase microextraction with in situ derivatization
followedbygaschromatography/mass

spectrometry”,

JournalofChromatography A, 2009, 1216, 5340-5346.
3. Qiaomei, Lan Zhang*, etc.

“Identification and quantitation of auxins inplants by liquid chromatography/electrospray ionization ion trap mass
spectrometry”, Rapid Communications in Mass Spectrometry, 2008, 22,2565-2572.
Lan Zhang* is the corresponding Author.

Break through point of micro GC and future image of advanced GC
T. Nakagama1 and K. Uchiyama2
1

College of Industrial Technology, Nihon University, 1-2-1 Izumicho, Narashino, Chiba, 275-8575,

JAPAN, 2 Tokyo Metropolitan University, 1-1 Minamiosawa, Hachioji, Tokyo, 192-0397, JAPANMiniaturized and microfabricated instruments for GC are of interest because of their verysmall size, weight, and resource requirements and are intended for on-site analysis [1]. These
instruments are under development in several laboratories and some of them are commerciallyavailable. The advantages of micro GC for monitoring volatile and semivolatile organic compounds,
relative to conventional sorbent tube or whole-air sampling methods followed by laboratory analysis,
include the following: reduced sample handling, which minimizes loss, degradation, or
contamination; greater sample throughput; and the potential for more timely responses tomeasurement results [2]. Where nonspecific detectors are employed, qualitative analysis is based
solely on retention time. Instruments employing GC with more sophisticated detectors, such as mass
spectrometers or ion mobility spectrometers, can provide more definitive identification of eluting
species at the cost of increased size, power, complexity, and cost.
In our research, a compact GC by applying microfabrication and inkjet technologies forflow control, on-chip column separation [3] and liquid sample injection [4,5] was developed. A small
pencil-sized atomic emission detector (AED) using a helium radio-frequency microplasma was also
employed for the GC system [6]. This AED is capable of operating using a radio-frequency board of
up to 3W of applied power or with only carrier (helium) gas, and performed high selectivity andsensitivity by using a USB-driven CCD spectrometer. The detection limits of chlorine, bromine,
sulfur and phosphorus were obtained to be pg/sec levels. These GCs have a possibility of becoming
more compact and further improvements of their performance, robustness and usability will beeffective for saving of time, space, electrical power and cost for on-site and in-laboratory GC
analyses.
References[1] G. R. Lambertus, C. S. Fix, S. M. Reidy, R. A. Miller, D. Wheeler, E. Nazarov, and R. Sacks, Anal.
Chem., 77, 7563 (2005). [2] J. J. Whiting, C.-J. Lu, E. T. Zellers, and R. D. Sacks, Anal. Chem., 73, 4668
(2001).

[3] M. Nishino, Y. Takemori, S. Matsuoka, M. Kanai, T. Nishimoto, M. Ueda and K. Komori,IEEJ Trans, 4, 358 (2009). [4] T. Nishiyama, F. Endo, H. Eguchi, J. Tsunokawa, T. Nakagama, N. Seino,
M. Shinoda, T. Shimosaka, T. Hobo, and K. Uchiyama, Chem. Lett., 35, 272 (2006). [5] T. Nishiyama, F.
Endo, H. Eguchi, T. Nakagama, N. Seino, M. Shinoda, T. Shimosaka, T. Hobo and K. Uchiyama, Anal.

Sci., 23, 389 (2007). [6] N. Saito, T. Nakagama and K. Uchiyama, Bunseki Kagaku, 56, 729 (2007)(Japanese).

Biographical Sketch
Name : Tatauro Nakagama
Born : 1964

Nationality : JAPANAffiliation : Department of applied molecular chemistry, College of Industrial Technology, Nihon
University
Contact : E-mail: tatsurou.nakagama@nihon-u.ac.jp
Web:

http:// www.ic.cit.nihon-u.ac.jp/staff/professor/index28.html
http://kenkyu-web.cin.nihon-u.ac.jp/Profiles/IA/0007549/profile.html

Title : Associate Professor
Record :1989-1992

SANDOZ Pharmaceutical Co., Researcher

1992-2007

Tokyo Metropolitan University, Research Associate

2002-2005

National Metrology Institute of Japan, Research Scientist

2007-2008

Tokyo Metropolitan University, Assistant Professor

2008-

Nihon University, Associate Professor

Microfluidic chip for cell analysis
Wei Du, Pu Chen, Jian Sun, Bi-Feng Liu*
Britton Chance Center for Biomedical Photonics, Department of Systems Biology, College of Life Science& Technology, Huazhong University of Science and Technology, Wuhan 430074, China
E-mail: bfliu@mail.hust.edu.cn

Cell is the basic unit of life. To reveal cell function and its underlying molecular mechanism has beenhighly focused for a long time. With the development of cellular biology, new approaches are greatly
demanded for better understanding more and more sophisticated cell phenomena both in qualification andin quantification. Therefore, cell analysis becomes a major challenge for bioanalytical chemist to innovate
the techniques. Microfluidic chip, or also referred to as lab-on-a-chip is a new concept technology and isrecognized as the next-generation platform for chemical and biological analysis, due to its advantages ofminiaturization, integration, automation and high throughput. Because the size of microchannel inmicrofluidic chip is quite compatible with the dimension of a regular cell, microfluidic chip is an ideal toolfor cell analysis, which has been recently paid much attention.
In this work, we are trying to develop advanced microfluidic chips for multiple purposes of cell analysis.
1) Microfluidic chip for cell sorting: regularly, cell sorting is performed using flow cytometry. However,conventional flow cytometry is applicable only for a large amount of cells. Thus, we attempted to develop a
microfluidics-based cell sorter for sorting a small amount of cells. By using novel on-chip pressure gating
technology, the purity and recovery of sorting particles and cells are over 90%, revealing a better
performance than previous reports. 2) Cell stimulation on a microfluidic chip: Cell can give reply withvarious stimuli. Investigation on stimulation-reply can obtain valuable information about cell function. In
our work, we developed a microfluidics-based and time-controllable platform for cell stimulation. Cells
were cultured inside the microchannels. By using on-chip pressure gating-pinching technique, cell could be
stimulated and washed with a minimum time of 20 ms, and made the concept of the-same-cell-analysis
very applicable. Examinations of cell stimulations with ATP and histamine implied that our method is
simple, stable, accurate and reliable. 3) Microfluidic chip for cell clamping. We demonstrated a sidewall
clamping technique in microchannel, which could realize high throughput cell clamping, hence paved a
new approach for high throughput and automatic patch clamping. 4) Microfluidic chip for cell-to-cellcommunication. When cells were cultured inside the microchannel, a cell could be stimulated by chemicals
while other cells could not because of laminar flow effect in microchannel. With this way, we successfully
visualized the calcium waves among cells due to cell-to-cell communication by gap junction. To aconclusion, microfluidic chip is suitable platform for cell manipulation and analysis. Further applications oflab-on-a-chip technology on cell analysis are greatly promising.

Biography

Dr. Bifeng LIU
Professor, Chair
Department of Systems Biology
College of Life Science & Technology
Huazhong University of Science and Technology (Eastern Campus)
Wuhan 430074, China
Tel: +86-27-8779-2203 (Office)Fax: +86-27-8779-2170 (Office)
Email: bfliu@mail.hust.edu.cn

Prof. Bifeng Liu was born in1972 in Hunan Province, China. He received his BA in 1994 at
Hunan Normal University with a major in chemistry and PhD in 1999 in analytical Chemistryat Wuhan University (China). He then did a postdoctoral research in College of Life Scienceat Wuhan University. From 2001, he visited Prof. Shigeru Terabe’s group at Himeji Instituteof Technology (Japan) as a JSPS postdoctoral fellow. In Fall of 2003, he moved to Huazhong
University of Science and Technology (China), where he is now a full professor and serving
as the Chair of Department of Systems Biology. Dr. Liu currently is serving as the editorial
board member of several journals such as The Open Analytical Chemistry Journal, Journal ofComputer Science and Systems Biology, Chinese Journal of Analytical Science etc.
His research focuses on systems biology-oriented analytical science in the areas of micro
separations, microfluidics and molecular imaging. He is the author and co-author of over 50
original research papers and 9 invited reviews published in international journals.

Studies on Microfluidic High-Speed Capillary Electrophoresis
Qun Fang, Ting Zhang, Yong-Qiang Cheng, Qing-Hu Lin, Ya-Ni Dong
Institute of Microanalytical Systems, Zhejiang University, Hangzhou, 310058, China.
In 2008, we developed a microfluidic picoliter-scale sample introduction techniquebased on a short capillary and translational spontaneous injection technique. With thenovel approach, a high-speed capillary electrophoresis (HSCE) system was built toachieve high-speed separations of FITC-labeled amino acids [1]. The HSCE system
was composed of a short capillary and an automated sample introduction system withslotted sample and buffer reservoirs and a computer-programmed translational
platform. The inlet end of the capillary was mechanically ground into a tapered tip.
Translational sample introduction was performed by linearly moving the platform,
allowing the capillary inlet to enter the sample solution, and then remove from it toachieve spontaneous sample introduction by capillary action. The platform
continuously moved to allow the capillary inlet to enter the buffer solution, and CE
separation was performed by applying high voltage between the buffer and waste
reservoirs. With the HSCE system, five FITC-labeled amino acids including arginine,phenylalanine, glycine, glutamic acid and aspartic acid were separated within 5.5 s in
15 mm separation length. High separation efficiencies ranging from 0.40 to 0.46 μm
plate height were obtained.
Recently, we applied this HSCE system in the separation of DNA fragments andproteins under capillary gel electrophoresis mode. In the HSCE system for DNA
fragments separation, ΦΧ 174-Hae III digest DNA Marker was separated within 100 s
using an effective separation length of 2.5 cm. Theoretical plate number of 603 bp
fragment was 3.2×106 /m. In the HSCE system for protein separation, five protein
standards including myoglobin, egg albumin, BSA, phosphorylase b and myosin wereseparated in 60 s with an effective separation length of 1.5 cm.

Reference
1. Zhang, T.; Fang, Q.; Du, W.-B.; Fu, J.-L. Microfluidic Picoliter-Scale Translational
Spontaneous Sample Introduction for High-Speed Capillary Electrophoresis. Anal.
Chem. 2009, 81, 3693-3698.

When prion drop into the picnic basket of analytical chemist’s: Surprisingproperties of optical isomers of proteins and novel detection methods*

Li Qiang Chen, Ping Ping Hu, Sai Jin Xiao, Cheng Zhi Huang

Education Ministry Key Laboratory on Luminescence and Real-Time Analysiss, Southwest
University, Chongqing 400715 P R China.

Prior to the discovery of prion, it was thought that all pathogens of diseases used nucleic
acids to direct their replication. However, the "protein-only hypothesis" which was based on theconversion of a normal cell-surface glycoprotein (PrPC) into a conformationally altered isoform(PrPSc), proposed that a protein structure can replicate without the use of nucleic acid.1 This was
initially controversial as it contradicts the so-called "central dogma of molecular biology", which
describes nucleic acid as the only central form of replicative information. That is why prion has
attracted great attention of biologists in recent years.2 However, what will happen when proin was
highlighted by analytical chemists?
Our present work is composed of three parts, namely discussion the properties of opticalisomers of pion proteins, exploitation the novel detection methods, development of a variety oflabeling and imaging techniques for prion protein in living cells.
It has stimulated long-standing endeavors to understand homochirality in life and the
production of enantiomeric excess in prebiotic organic molecules. We took prion protein as amodel to design a simple experiment to discuss the instinctive relationship between protein
configuration and the stirring direction of their culture medium. It was surprisingly to find that theconfiguration of the cellular prion protein is dependent on the stirring direction (clockwise or
anticlockwise). By simply controlling the stirring direction, we have obtained prion protein with
distinctive properties of optical isomers corresponded with the stirring directions, and D-aminoacid was speculated to be concluded in the protein of cultured by anticlockwise direction. These
results occur us to further explore the mysterious homochirality in life exactly how and where in
the cell the optical isomers of proteins occur.
To develop the high selective and sensitive methods for prion proteins detection, aaptamer-participated haprin structure was designed by employing cellular prion protein (PrPC) as amodel protein, and thus an aptamer-mediated turn-on fluorescence assay for proteins wasdeveloped. The designed aptamer-participated haprin structure consists of three segments. Namely,
an aptamer sequence located in the loop, three guanine bases at 3’-terminal, and a fluophor
modified at 5’-terminal. It was found that the guanine bases at the 3’-terminal could quench thefluorescence of the fluophor such as tetramethyl-6-carboxyrhodamine (TAMRA) at the
5’-terminal about 76.6% via electron transfer if the guanine bases are close enough to the fluophor,and the quenched fluorescence could get restored when the target protein is present since theinteraction, which could be confirmed by measuring fluorescence lifetime, between
TAMRA-aptamer and the target protein forces the guanines away from TAMRA so that
TAMRA-modified aptamer changes into turn-on state. We also developed a new ultra-sensitive
prion protein detection strategy that based on the fluorescent resonance energy transfer (FRET)
between PrPC functioned quantum dots (QDs) and nanometer-sized gold particle (AuNPs) labeledwith Anti- PrPC aptamer. The detection limit was low to 1.0 fg/mL.
*

Corresponding author. Tel.: +86-23-68254659; Fax: +86-23-68866796. E-mail address:
chengzhi@swu.edu.cn (C.Z. Huang).
1Aptamer adapted gold/silver nanoparticles (Apt-G/SNPs) probes with both stability andbiocompatibility were developed as a novel optical probe for labeling and imaging of prion protein,
wherein G/SNPs act as illuminophore and aptamer as biomolecule specific recognition unit,
respectively. It’s further application demonstrates that the caveolae-related endocytosis is likely anecessary pathway for Apt-G/SNPs labeling PrPc internalization in SK-N-SH cells. The integrated
capability of Apt-G/SNPs to be used as light scattering and TEM imaging agents, along with theirpotential use for single particle spectral analysis, makes them a great promise for future
biomedical imaging and diseases diagnosis. For real time labeling and imaging of prion protein in
living cells, aptamer conjugated QDs as an optical probe was also designed by employing
aptamers as “underdiscovers”. The utility of this probe has been demonstrated in real time
tracking of cellular prion protein.
Keywords: prion, optical isomers, aptamer, detection methods, optical imaging probes
References1.

Prusiner, S. B., Proc. Natl. Acad. Sci. USA 1998, 95, (23), 13363-13383.

2.

Aguzzi, A.; Polymenidou, M., Cell 2004, 116, (2), 313-327.

2

A Study of Gold-Coated Magnetic Nanoparticles for Solid-Phase Immunoextraction:
Determination of Epitestosterone in Urine by High-Performance Liquid
Chromatography

Shuang Qiu, Li Xu, Yi-Ran Cui, Qin-Pei Deng and Xin-Xiang Zhang*
Beijing National Laboratory for Molecular Science, College of Chemistry, Peking
University, Beijing 100871, China* To whom correspondence should be addressed. E-mail: zxx@pku.edu.cn. Tel:
+86-10-62754174. Fax: +86-10-62751708.

A solid-phase immunoextraction method for quantitative analysis of epitestosterone(ET) in urine samples by high-performance liquid chromatography (HPLC) has beendeveloped based on gold-coated magnetic nanoparticles. In this mothed, half-IgG
fragments of anti-ET monoclonal IgG antibodies were covalently immobilized onto
magnetic core-shell Fe3O4@Au nanoparticles. The magnetic nanoparticles (MNPs)
were used as a solid phase for extracting epitestorsterone in spiked human urine
samples. An external magnetic field was applied to recover the MNPs which werethen washed with distilled water followed by eluting with absolute methanol to obtain
ET as the analyte. The obtained extraction solution was analyzed by HPLC with UV
detection (244 nm) within 12 min. The standard calibration curve obtained for ET
showed good linearity in the range of 20-200 ng/mL in phosphate-buffered saline

1

(PBS) solutions with acceptable accuracy and precision results. This method has anenrichment factor of 100-fold to fulfill the need for measuring analyte with its
concentration 100 times lower than the limits of quantification (S/N = 10). The results
obtained by the present method for spiked urine samples were in good agreement withthose from indirect competitive enzyme-linked immunoadsorbent assays (ELISAs).
The appoarch described here is high specific, fast, easy to operate and is likely to havethe advantage of conventional immunoaffinity column technique.

Acknowledgement: This work was supported by National Science Foundation ofChina(No90713013)

and

the

China(2006BAF07B03,2009CB320305)

2

Ministryof

ScienceofDevelopment of advanced ion chromatography applied todeveloping ocuntries

R. Goto1
1

Summary

References[1]Biographical Sketch
Name : Ryozo Goto
Born :
Nationality : JAPANAffiliation :Contact : E-mail:
Web: http://www.
Title :
Record :A Novel Approach for Aroma Components Analysis Using
Monolithic Material Sorptive Extraction method.”MonoTrap”
Atshushi Sato1, Katsuhiko Sotomaru, Manami Takeda2
1

Department of R&D,GL Sciences,273-2,Sayamagahara,Iruma,Saitama 358-0032,Japan

Summary
Monolithic Material Sorption Extraction (MMSE) is a novel approach for sample adsorption andextraction using a monolithic hybrid adsorbent -MonoTrap. MMSE has several advantages such as
effective pretreatment with simple operation and high-efficiency adsorption capacity. In this study,
Various Aroma components were analyzed by MMSE and SBSE method.
Key word: MMSE, MonoTrap, Aroma, juice, Food, Coffee.
1 Introduction
A Conventional media for sample adsorption and stir bar sorptive extraction (SBSE) and solid-Phase
micro extraction (SPME) are coated with an extracting polymerphase, for example, polydimethylsiloxane(PDMS).Disadvantages of conventional media are their small surface area and thin polymer phase coating
resulting in low recovery and need for a long extraction time and long conditioning time. The newly
introduced product, MonoTrap is a new generation media for adsorption and extraction developed byusing silica monolith technology. The state-of-the-art silica monolith technology allows us to manufacture
an innovative hybrid adsorbent of silica and activated carbon (graphite carbon) having a large surface area
bonded with octadecyl silane (C18). This C18 bonded hybrid medium shows quite effective adsorption
capability to a wide variety of compounds. In this study ,the aroma component of peach juice wereanalyzed by MMSE and SBSE methods.

2 Materials and Methods
Instrumentation. Analyses were performed GC2010 and GCMS,QP2010 (Shimadzu) ,Thermal desorption unit (T-Dex) and GC Capillary Column IC-PureWAX(30mX0.25mmi.d.,df=0.25um),MonoTrap-TD(*2)(graphite carbon) (GLSciences).
3 Result and Discussion
As a result, it was confirmed that MMSE

(x10,000,000)
5.5

has the capability of the Adsorption the

5.0

Some Aroma components far better than

4.5
4.0

MMS

3.5

SBSE method.

3.0

An component with high sensitivity is

2.5
2.0
1.5

SBSE

1.0
0.5

described below.
1;beta-Linalool、2;gamma-Caprolactone
3;alpha-Terpineol、4;Benzyl acetate

0.0
-0.5
5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

50.0

Corresponding author, Name, E-mail Address

55.0

4 References[1] C.L. Arthur, J. Pawliszyn, Anal.Chem.,1990,62,2145[2] E.Baltussen , P.Sandra ,F. David, C.A.Cramers., J.Microcol.Sep.,1999,11,737[3] C.Bicchi,C.Iori,P Rubiolo,P.Sandra., J. Agric. Food Chem., 2002, 50 (3), pp 449–459[4] E.E. Stashenko,J.R.Mart nez., J. Sep. Sci. 2008, 31, 2022- 2031
[5] Gerstel Solutions Worldwide-May 2007
5 Biographical Sketch
Name : Atsushi Sato
Born : Japan Fukushima Prefecture Fukushima City
Nationality : Japan
Affiliation : Department of R&D,GL Sciences
Contact : E-mail: a.sato@gls.co.jp
Web: http://www.
Title : A Novel Approach for Aroma Components Analysis Using
Sorptive Extraction method.”MonoTrap”
Record :Corresponding author, Name, E-mail Address

Monolithic Material

The recent trend of analysis method for food safety in China
Yuki Hashi (Ph.D.)
Shimadzu International Trading (Shanghai) Co. Limited
E-mail: y-hashi@shimadzu.co.jp
In recent years, public and scientific concern about food contaminations has been increased seriously due
to their potential risks to human health. The monitoring of the contaminants such as pesticides andantibiotics in agricultural products and process foods, and intentional addition of illergal hazardous
substances such as melamine and cyclamate etc. in food stuff must be carried out before these products are
released to the market. However, high throughput analysis is biggest challenge to these products since
freshness of these products are necessary. Moreover, the cost of analysis is also one of importantconsiderations for development of the new analytical method. Although the performances of the analytical
instrument for ultra fast determination have rapidly been improved in recent years, the sample pretreatment
procedures remain the bottleneck for rapid analysis. Therefore, economical and effective pretreatment
methods to eliminate matrix interferences, to achieve high throughout are demanded.
Recently, solid phase extraction (SPE) is getting most popular technique as a replacement of classicalliquid-liquid extraction (LLE) in China, since SPE has the advantages of quick treatment and affordable
cost. Most of the latest analytical methods such as residual pesticides analysis or melamine analysis areincluding this SPE procedure. However, the cost of SPE tube is not really cheap for the routine analysis
purpose. Therefore, we have introduced matrix solid-phase dispersion (MSPD) for the determination ofresidual pesticides in vegetable and fruits. Same sorbents used for SPE can be dispersed to homogenizedmatrices for solid sample or liquid sample for extraction and clean-up. In order to improve the extraction
efficiency for polar compounds, large amounts of Na2SO4 are usually added in this procedure. The
combination of MSPD and on-line sample pretreatment GCMS instrument such as GPC-GCMS
hyphenated system is quite acceptable in China market due to low cost and quick determination for residual
pesticides.
In this report, the recent trend of analysis method for food safety in China including latest Chinese official
method, residual pesticides analysis and melamine analysis will be introduced.

Biographical Sketch
Name

Yuki Hashi, (Ph.D.)

Title

Shimadzu International Trading (Shanghai) Co., Limited
Shimadzu Shimadzu Global COE for Application & Technical Development
General Manager, Chief Scientist

Address Shimadzu International Trading (Shanghai) Co., Limited , Shanghai Office
Block E, No.570 West Huaihai Road, 200052, Shanghai, China
Phone: +86-10-8525-2352
Fax: +86-10-8525-2330
E-mail: y-hashi@shimadzu.co.jp
http://www.shimadzu.net.cn
Birthday27th February, 1961

Nationality Japan
Research Major
Dr. Hashi graduated from Dept. of Chemistry, Osaka University, Osaka, Japan in 1986. At thesame year, he joined Shimadzu Corporation, analytical instrument division to develop HPLC
instrument especially electrochemical detector and column packing materials for protein
analyses. In 1994, he moved to Shimadzu Singapore as an application manager for
chromatography instruments to take care of the customers in South-East Asia as well as south
Asia countries. His major area of interest is development of new application system using LC,
GC, LCMS and GCMS for the enhancement of analysis efficiency. Based on this researchwork, he has received a Ph.D. degree from Research Center for Eco-Environmental Sciences,
The Chinese Academy of Sciences in 2009. Currently, he is interested in themulti-dimensional GC/GCMS system for expansion of applicability. Application area is not
only for food safety but also, for pharmaceutical, environmental and life science such as
genomics and proteomics. Beside his research work, he is also actively providing the lectureof basic instrumentation in university, and contributing his papers to some technicalconferences.

Food Regulation and Analytical Method for the Food Safety in KOREA
Jaeho HA, Ph.D.
Food Analysis Center, Korea Food Research Institute,
Bundang, Seongnam, Gyeonggi Province, Republic of KOREA
Tel : +82 31 780 9127, Fax : +82 31 780 9280, E-mail : jhkfri@kfri.re.kr
Food safety issues have been attracting a big concern in every country, becoming global
problems in a very short period soon after its outbreak. Over the past 10 years, a lot of food safetyissues have been attracting the national and international interest. The consumers require eating thesafe and wholesome food, while the government, producers, manufacturers and research instituteendeavor themselves to find the best way providing the safe food to the people. Based on our pastexperience, providing safe food to the consumers is considered to be one of the most difficult tasks to
be solved.
Safe food must be free from the following hazards to health; Biological Hazards (Pathogenic
bacteria, virus, parasites, worms); Chemical Hazards (Natural toxins, agricultural chemicals,
environmental contaminants, food additives); Physical Hazards (Stones, metal fragments, boneshards)To provide the safe food to the consumers, the effective food control system is required ingovernmental and private sectors. Where food safety is concerned, most countries share the objectivesof protecting consumers from health hazards and commercial fraud. Achieving these objectives
require a food control system based on a fundamental food law, accompanied by detailed regulations
and administered by an efficient food control organization. Therefore when food legislation is enacted
the importance of consumer participation must be recognized, as this will enhance consumer
confidence in the national food control system.
On the other hand government food control agencies tend to avoid consumer organisations,which they view as antagonistic and not representative of the true concerns of the individual consumer
within the country. In the eyes of the of government officials the issues raised by consumer groups
seems irrelevant and even when they are viewed as important there may not be enough resources todeal with the issues that were raised.
Analytical method to ensure the food safety, food sanitary law related with food and foodadditives in Korea include the food code, functional food code and food additive code. In the foodsanitary law, the analytical methods for proximate components, hazardous materials, nutrients,
functional compounds, food additives and other components. In the presentation, food regulation andanalytical method in the view point of food safety has been discussed.

韓国の環境政策と環境分析研究の最前線
玉韓国坤

国立釜慶大学校環境大気科学科、地球環境工学大学院、ダイオキシン研究センター
Gon OK

Department of Environmental Atmospheric Science, Graduate School of Earth Environmental
Engineering, and Dioxin Research Center, Pukyong National University

SummaryThis paper describes the historical development of Korean environmental analysis research, and theenvironmental issues and policy in Korea. In 1950s, DDT and synthetic chemical compounds were usedduring the Korean War. 1960s and 1970s were the time of industrialization in Korea, which began tobring up the pollution issues. The government and people of Korea started to experience the change intheir concept of environmental issues between 1980s and 1990s.

In 21st century, environmental policy

and laws were prepared as counter plans for the global environmental issues.
The Environmental Analysis field showed the change in trend from criteria pollutants to toxic compounds
like Hazardous Air Pollutants (HAPs) and Persistent Organic Pollutants (POPs), and to trace level
analysis and environmental problems.
The monitoring data of environmental analysis is the important scientific evidence of risk assessment of
humans and ecosystems. The results of these monitoring data are being used to reduce the human andenvironmental risk, to create safe living environment and to find solutions for the global environmental
issues. Thus, it can be concluded that “Sources and Environment Budget System Approach”, which
begins from the discovery of the source to the environmental media, should be completed for the futureenvironmental analysis research.
Key word:

Dioxins, Emerging POPs, HAPs, Sources Inventory, Environ Budget System

環 境 汚 染 究 明 と環 境 分 析 化 学
地球環境５６億年の変化の中で人類が生き残るため活動してる中で作り出した化学物質の種類を明確に答えることは出来ないと思われる。近年、環境問題は地域的問題ではなく、また、一つの国に留まる問題でもない地球規模の問題として取り上げられている。 それは産業発展と共に作り出した化学物質の製品だけではなく、製品の生産などの過程で非意図的に発生した化学物質
が環境汚染物質として越境移動している問題を乗り越え両極地までの影響を与える状況である。
これらの現状を分かるようになった科学的な研究成果の基盤は化学分析研究者の努力に伴なう分析技術の発展の成果の一つである。

その歴史の中で地球環境５６億年の生命誕生のMillerの

地球大気再現実験などの科学的根拠を作り出したこともクロマト分析化学であり、成層圏のオゾ

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

ン破壊問題を指摘していたRowland 教授とMolina博士の研究の基盤にはECD-クロマトグラフの開発のより1970年代のLovolock博士による大西洋大気中のCFCsの分析結果によることである。
また、近年の環境ホルモンの問題、ダイオキシン類の問題、残留性有機汚染物質（POPs)等のその出発と原因の解明、対応など研究の中心にはクロマト分析技術が大きな役割を果たしてきたと言われることが出来る。
全世界におけるこれらの分析技術の発展の中で、特に、環境分析研究は一般の化学分析とは異なり極微量であり、環境媒体からの数多くの化学物質の中での目的成分の同定の分析技術の研究能力が要求されていた状況であった。これは環境分野研究においての特殊性の一つであり、環境モ
ニトリングは発生源、大気などの環境プロセス過程の媒体、水、土壌、低質、生物などの環境収
容体と人における暴露の過程までの喪失と濃縮の繰り返し現状を持ち、自然の中での複雑な過程の解明作業である。これらの特殊性を持つ環境試料の分析結果は国の政策と社会的ないろいろの面との関係を持っている。 その結果は政策に対し一つの尺度的であるため科学的な根拠を持ちいる事が要求されていると共に法律的、行政的実行の根拠として用いられ環境管理の政策の作りと実行の科学的な出発が環境分析の位置付けである。

韓国の歴史的な環境問題と環境政策の法律的根拠
韓国の場合でもこれらの環境分析の地位と言うものは世界共通のことである。 韓国の環境管理の政策も同じく、近年の国際レベルの分析技術に基ついて科学的な根拠により法律と行政実行の流れを持ち行けるように成りつつある。
そして、韓国の環境管理の政策において法律的な根拠は以下のようなことにある。


憲法第35条: すべての国民は健康で、快適な環境で生活できる権利を持ち、国家と国民
は環境保全に努力すべきであると述べられている。



この意味は広義では、環境政策基本法第3条1項において規定している“自然環境と生活環境”に関わる関連の法律である。しかし、狭義では、政府の組織の環境部に関わる事として自然環境と生活環境の保全と環境汚染防止に関する法律である。

このことで、韓国の環境問題に対する政策の歴史的な流れを見ると、まず、1950年6月25日に起きた韓国戦争は国土環境と経済の破壊、苦しい国民環境生活などの様々な問題問題が生じていた。それに対し、国の再建設のため産業の発展は国家最大の目標でもあった。1950年代の韓国戦争はまず、生活衛生問題を起こし、合成化学物質に依存するきっかけになり、最近のPOPsの一つであるDDTという化学物質の本格的な使用の始まりであったかも知らない。韓国においての忘れられない６．２５の韓国戦争はDDT使用による“奪われし未来”の出発であったかも知らない。 韓国戦争が終わり、国家の再建設は国の第一の目標であり、工業を中心とした産業経済活性化の 第１次経済開発５階年計画による工業化の推進が行ったのが1960年から70年代であった。それに伴ない公害対策として環境管理の政策の始まりが1960年代韓国の環境政策は公害対策であった。

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

図

韓国戦争の被害と女優のマリリンモンロのアメリカ軍人の慰問講演の訪問

図

アメリカ軍による北朝鮮軍と子供にDDTの散布

ʻ第１次経済開発５階年計画による工業化の推進が１９６３年に公害対応の環境基本法である「公害防止法」が制定·公布された。



しかし、公布された公害防止法は予算を含め実施に伴なう法律がまとまらなく、その役
割は微々の状況であった。



1960年代と70年代に掛け繊維、衣類、製鉄、化学、造船など重化学工業を中心とした経済開発が本格的に行い蔚山―温山地域などで発生した環境汚染問題は恩山病の名前が付けられ大きな社会問題として取り上げられていた。

韓国の産業公害から環境問題までに関わる環境分析の話題
韓国の産業発展とこれらの公害汚染問題からの分析対象の汚染物質は主に,重金属、窒素酸化物
質(NOx）,黄酸化物(SOx),塩化水素（HCl）などが発生源と環境中の主なモニトリングの対象物質であった。特に、大気汚染のモニトリングではダストージャ方法で亜硫酸カスのモニトリングをしたり、発生源の煙突排ガスの測定のため一日4個位の煙突を昇った記憶がある。そして、機器分析は主にAAとUVが使用されていた。
また、温山の公害問題以前の７０年代半ばには、海洋汚染の一つとして国レベルで大きく取り上げられた分析研究結果としては当時、韓国釜山水産大学の元ジョンフン教授により韓国の南海岸

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

の水産物の重金属汚染の研究結果が日本の“ぶんせき”学会論文に報告され、韓国の水産物の日本に輸出が出来なくなることの事件が起きた。その時代の韓国経済成長に水産分野の役割は大きく、それに影響を与えたことで朴大統領から大学は重い責任をとることが指摘されたが研究者を育つことも大切であったことも認めたと言うエピソドがあった時期でもある。しかし、その時代、
朴大統領は韓国国土の緑化、し尿などを用いエネルキー化など最近の地球温暖化問題に対応出来る環境政策を世界一に取り上げ未来を設計していた政治家であったと思う。
１９７０年代後半、「公害防止法」が廃止され、この代わりに「環境保全法」が1977年12月31日
制定·公報され環境基準及び排出基準、環境監視制度、防止施設の設置の義務化と環境影響の評
価などの一連の根拠が作り出された環境政策の基盤が出来た時期であった。
1980年から1990年代は韓国の産業の発展がもっと進められ従来の産業汚染と自動車排ガス、都市地域の下水、ごみ問題、自然生態系の破壊を始め生活消費と関係する環境問題が大きくなり始
めていた。
1990年から2000年には本格的に公害汚染から環境汚染の問題へに転換されると共に汚染の対象物質は無機から有機の汚染物質の問題が取り上げられるようになった。特に1990年代に入り
都市大気の空気質の悪化、有害化学物質の使用の増加、廃棄物排出の増加などにより埋立地などの最終処分場の確保の問題の発生から廃棄物の焼却によりダイオキシン問題が1995年本格的に取り上げられるようになった。それの事で国レベルで、1996年環境部からの研究依頼を受け廃棄物焼却によるダイオキシンの排出実態の調査と排出ガイドライン作りの研究が釜慶大学の私の研究室で本格的に始まることになり、ごみ焼却からのダイオキシン対策に対する政府政策が取りまとめられた。
2000年代に入り、環境政策は事前予防の原則（Precautionary Principle）に伴ない政策を検討するようになっており、環境部が管理する関連法律は４６に拡大された。また、大気と水質に関して汚染物質の環境排出に対し総量の管理制度が導入されると共に、水に関しては使用者の負担原則と廃棄物管理においては生産者責任の制度が実行される時代になった。
特に、2000年には釜慶大学ダイオキシン研究センターの黄砂中ダイオキシンモニトリング研究結果からダイオキシン類の越境移動している問題が最初に取り上げる事にもなり春の黄砂の環境問題が社会問題として関心を高まることになった。

図

黄砂とダイオキシン類の越境移動

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

そして２０００年代での環境政策はグロバル化が進んであり、残留性有機汚染物質（POPs）管理を目的として2007年 残留性有機汚染物質（POPs）管理法が制定された。

韓 国 の ダ イ オ キ シ ン 類 な どPOPsの 対 策 と モ ニ ト リ ン グ
これらの法律を行かせるためには

まず、発生源の目録化により排出量を把握する必要があり、2001年にダイオキシン類（PCDD/DFs及びCo-PCBs）に対し、国レベルのダイオキシンインベント
リのカテゴリ分類など方法を環境部から依頼され釜慶大学ダイオキシン研究センターと環境管
理公団が共同に開発し2001年から発生源のモニトリングが始まり2005年に排出量がまとまるようになった。図に韓国のダイオキシンインベントリ推進の概要を表した。韓国のダイオキシン類のインベントリ分類カテゴリは発生量が多く予想される順位になっており、それに伴ない発生源のモニトリングの優先順位と削減対策の優先順位の位置付けが決まる事になった。

図

韓国のダイオキシンインベントリ推進の概要

また、韓国と外国とはの違いのもう一つの特徴は発生源の中での火葬場において外国には廃棄物のカテゴリに分類されているが、韓国の場合は廃棄物とは区別し、別のカテゴリに分けることにした。これは人間尊重の韓国の精神と文化の哲学の概念を用いる事にしたのが理由である。これ
らのインベントリ計画により発生源のモニトリングが実施されダイオキシン類の削減戦略が本格的に実行されることになった。

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

表韓国のダイオキシン類のインベントリの分類カテゴリ
Categories
Main

Semi-subSub

Waste Incineration

Municipal, Industrial, Hazardous

1. Waste Treatment
2. Ferrous Metal
Production
3. Non-Ferrous Metal
Production

4. Production of mineral
Products

5. Production ofChemicals
6. Power generation

Landfill

Controlled, Non-controlled

Iron and Steel Production

Sinter Furnace, Electronic Arc
Furnace, AlloyFoundries

Pig-iron, Steel Casting

Smelting, Refining and Alloy
Rolling,
Compressing
elongating

Copper, Aluminum, Lead and Zinc
andCopper, Aluminum, Other

Non-Ferrous Metal Foundries

Aluminum, Copper, Other

Glass

Flat Glass, Industrial Glass

Ceramic

Refractory, Non-refractory

Cement and Lime

Cement, Lime

Other

Asphalt

Chloride Compounds

EDC, VCM, PVC

Pulp and Paper Manufacturing

Pulp and Paper

Power generation
Area heating Facility

7. Combustion of
Manufacturing Industry
8. Heating

Furnace of Process
Commerce and Public Facility
Residence facility

9. Transport

Gasoline, Diesel, LPG

10. Others

Crematory, Forest Fire, Fire
accident, Tobacco

そして、２００５年にはダイオキシン類以外の農薬類POPsの公定試験法が環境部からの依頼を受け検討されるようになった。その対象の環境媒体は

まず、大気、水、土壌、低質を中心にし、試料の採取方法を含む、高分解ガスクロマトグラフ・高分解質量分析機（HRGC/HRMS）を用いた試験法が制定された。
韓国の環境中のPOPsの中でダイオキシン類はすべての環境媒体から検出されているが、農薬類のPOPsはダイオキシン類に比べ検出頻度はかなり少なく、農薬類POPsの中ではDDTとLindanが別の化合物に比べ検出される頻度があるが、その濃度レベルも外国に比べかなり低いことも確認された。
また、海洋汚染に対しては韓国の西海の低質からのダイオキシン類の調査の結果が１９９９年発表された以降、海洋汚染の現状の把握のため海洋水産部からの研究計画を作り環境ホルモンとPOPsに関する調査研究が２００２年から始まりダイオキシン類を中心に全国モニトリングが行っている。 これらの一連の研究結果らは２００６年国レベルで環境残留と環境暴露評価の報告がまとまり、焼却からの発生は２５０〜２５７ｇ WHO –TEQ/year、製鉄などを含む非焼却源からは１５４ ｇ WHO –TEQ/yearで推算された。また、一般の生活環境において食品を含め環境

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

経路からの暴露のレベルは０．５０５pg WHO-TEQ/kg/dayとして推算された。
また、近年UNEPは１２グルプのPOPs以外、PBDEsなど９種のEmerging POPsが追加された。韓国でも有害汚染物質管理及び残留性有機汚染物質（POPs）においての政策はダイオキシン類を出発にして Emerging POPs としてのPBDEs、PFOS/PFOAなどのモニトリングが行われている。
その結果を見ると先端IT産業の環境問題が新しい局面の状況にあると考えられる。

大気中の有害化学物質のモニトリング
もう一つの環境問題は化学物質の移動及び排出の概念であるTRIからの情報と国際的な管理が要
求 さ れ て い る REACH(Regulation on the registration, evaluation, authorisation andrestrictions of chemicals) などからも新しいアプロチが要求されている。
そして、有害大気汚染物質（HAPs; hazard air pollutants）により人のリスクの削減の対策が要求され,

２００８年HAPsインベントリを検討し化学関連の産業施設を始めHAPsの発生源からのモニトリングを行っている。
HAPsインベントリのモニトリングには対象になる化合物の種類が５３化合物であり、無機と有機化合物に対する試料の採取と分析が必要である。このモニトリングにおいて試料の採取方法と分
析方法は５グルプ以上が要求されている。特に、分析にはガス状の試料を吸着材等を用い直接に機器分析に対応できるGerstel-TDS システムとGC/MSDを組み合わせて分析などが主に用いられた。
また、２００１から２００２年には温室ガス対策のため排出インベントリも検討された。 分析の対象化学物質は主にCO2, CH4, N2Oが取りあげられているが一般的には電気化学センサーの連続ガス分析とガスクロマトグラピのGC-FIDとGC-ECDが用いられたが、それと共に我々は特に N2Oに対してはGC-MSDを用い確認と分析結果の精度を上げる事にしていた。

終わりに最近の韓国での環境に対する分析の対象は微量で、毒性が強く、人をはじめ生態系に与える悪影
響を及ぼす可能性が高い化学物質の研究が活発に行われるようになった。また、環境分析はただの規制基準に基ついたことでなく、環境の管理に事前対応出来、新しい化学物質の管理に必要な科学的な解明の研究が進んでおり、その中の中心に微量で、制度よい環境媒体を対象にしている
化学分析の役割が存在し、環境媒体の微量化学物質の分離と同定にクロマト分析の大切さが目立っていると言われる。
その環境分析結果はただの濃度と規制の意味ではなく、人と環境生態系の管理とリスクアセスメ
ントの大切なパラメタ‐作りの資料としての活用につながっている。人と環境のリスクアセスメ
ントを行うためには発生源から環境媒体のプロセス全般と食品、生物、人の暴露試料まで分析が要求されている。
したがって、空から海の下まで分析は幅広くアプロチをし“Sources and Environ Budget
System Approach ” の研究シナリオによる環境研究は必要であると思う。

Corresponding author, Gon Ok, E-mail Address:gonok@pknu.ac.kr

The Analytical Chemistry Division on Chinese Chemical Society andRecent Research Topics in China
Jin-Ming Lin
Department of Chemistry, Tsinghua University, Beijing 100084, China
E-mail: jmlin@mail.tsinghua.edu.cn
The Chinese Chemical Society (CCS) is an academic organization formed by Chinese Chemists of theirown accord and aiming at uniting Chinese Chemists at home and abroad to promote the development ofchemistry in China.
The CCS was founded in Nanjing (Nanking) on August 4, 1932. Since then sixteen Boards of Directors
had been elected by 1949 with more than 3000 individual members and 150 group members. In 1959, the
CCS merged with the Chinese Chemical Engineering Society to become the Chinese Chemical andChemical Engineering Society, but was split up again into two separate organizations, the Chinese
Chemical Society and the Chinese Chemical Engineering Society in 1963.
At present, there are more than 55000 registered individual members, some foreign members and 58
group members. Under the CCS there are 7 Academic Committees, 15 Specialized Committees and 7
working Committees. The CCS has established a close relationship with some chemical organizations in
the countries from Europe, America, and Asia. Since 1983, the CCS has set up 11 kinds of awards,
including Young Chemists Award, Professor Wang Baoren's Award for accomplishments in the field ofpolymer science, Professor Liang Shuquan's Award for accomplishment in basic research in analyticalchemistry, Chuangxin Award for Polymer Science etc.
The Analytical Chemistry Devision is one of the 15 specialized committees of CCS. Prof. Erkang Wangis the chairman. Prof. Jin-Ming Lin and other six professors from different universities or institutes are theassociate chair for the devision. Each three years, we have once national symposium on analyticalchemistry. This year during October 29 to November 2, the 12th National Symposium on Analytical
Chemistry will be hold at Yangzhou City. More than 1000 analysts and students will attend thissympposium. Liang Shuquan’s Award and the Awards for the Excellent Research Papers published in Fenxi
Huaxue (Chin. J. Anal. Chem.) during 2005-2007 will be announced at the conference. Beside this
symposium, each year there are many different meetings for the different research topics, for example, life
analytical chemistry, luminescence, capillary electrophoresis, μ-TAS, spectrometry, electroanalyticalchemistry, chromatography, mass spectrometry, environmental analytical chemistry, sensor, etc. In thepresent presentation, I shall introduce the function of the Analytical Chemistry Devision of CCS and therecent resrach topics in China. I hope to be as a bridge between China and Japan in the research area ofanalytical chemistry.

Biography
Professor Jin-Ming Lin (Ph.D)
Professor of Cheung Kung Scholarship, Ministry of Education, China
Vice Chair, Department of Chemistry, Tsinghua University, Beijing 100084, China
Tel/Fax: +86-10-62792343
E-mail: jmlin@mail.tsinghua.edu.cn

Professor Jin-Ming Lin was born in 1963 in Fujian, China. He received his BA degree in 1984
and received a PhD degree in analytical chemistry at Tokyo Metropolitan University in 1997.
He had studied and worked in Showa University and Tokyo Metropolitan University during
1992-2002. He is currently a full professor of Tsinghua University since 2004. He is a vicechairman of the Division of Analytical Chemistry, Chinese Chemical Society, and a memberof council of Chinese Society for Chromatography Science. He is a contributing editor ofTrends in Analytical Chemistry, a guest editor of Talanta for special issues ofChina-Japan-Korea Environmental Analytical Chemistry and editorial broads of Talanta,
Luminescence, Eurasian Journal of Analytical Chemistry, J. Flow Injection Analysis,
International Journal of Analytical Chemistry and seven Chinese journals. He received several
awards for his contributions in chemiluminescence and separation science: Outstanding
Young Chemist Award (Chinese Chemical Society, 1992), Young Analyst Award for Flow
Injection Analysis (Flow Injection Section, Japan Society of Analytical Chemistry, 2000) ,Kanton New Century Award (Japan Society of Analytical Chemistry, 2001), National
Science Fund for Distinguished Young Scholars of China (National Natural Science
Foundation of China, 2002), FIA Award for Science (Flow Injection Section, Japan Society ofAnalytical Chemistry, 2008) and GC Contribution Award (GC Discussion Group, Japan
Society of Analytical Chemistry, 2008)
His current research is focused on sample pretreatment, chemiluminescence, capillaryelectrophoresis and microfluidic device. He is the author and co-author of 218 original
research papers published in international journals, 21 reviews, 3 books and 31 patents.