4th EucheMs chemistry congress

wednesday, 29-Aug 2012
s606
chem. Listy 106, s587–s1425 (2012)
Analytical chemistry Electrochemistry, Analysis, sample manipulation
spectrometric methods – i
o - 2 5 5
LuMinoL CheMiLuMineSCenCe deteCtion of
AMino ACidS with enzyMAtiC reACtionS
A. KuGiMiyA 1 , t. hAMAoKA 1
1 Hiroshima City University, Center for Industry and Public
Relations, Hiroshima, Japan
The determination of amino acids is greatly important in
clinical settings and food technologies such as the measurement
of diagnosis of diseases in clinics and the measurement
of freshness and taste of foods in food technologies. The
conventional methods of detection for amino acids are pre-column
labeling for High-Performance Liquid Chromatography (HPLC)
of amino acids with fluorescence derivatives, or post-column
labeling for HPLC with ninhydrine, but large amounts of organic
solvents and an experimental step for labeling amino acids are
necessary.
In this study, for the rapid and easy measurement of amino
acids, luminol chemiluminescence detection method of amino
acids was developed with enzymatic reactions. Aminoacyl-tRNA
synthetases (ARSs) were used as the molecular recognition
element for amino acids. The biosensing system for asparagine
was constructed with asparaginyl-tRNA synthetase (AsnRS) for
recognition of asparagine and the luminol chemiluminescence
intensity was measured by way of several enzymatic reactions.
By using this sensing system, up to 200 μM of asparagine could
be measured selectively.
This work was partly supported by the Hiroshima City
University Grant for Special Academic Research (General Studies).
Keywords: amino acid; biosensor; protein; chemiluminescence
detection; enzyme;
spectrometric methods – ii
4 th EucheMs chemistry congress
o - 2 5 6
intrA-oPerAtive APPLiCAtion of viBrAtionAL
SPeCtroSCoPy
r. SALzer 1 , G. Steiner 2 , M. KirSCh 3
1 Dresden University of Technology, Department of Chemistry
and Food Chemistry, Dresden, Germany
2 Dresden University of Technology, Faculty of Medicine
Clinical Sensoring and Monitoring, Dresden, Germany
3 Dresden University of Technology, Faculty of Medicine and
University Hospital Neurosurgery, Dresden, Germany
Various techniques are established for biochemical
characterization of tissue and for clinical diagnosis, several other
methods based on new technology are under development. [1]
Unfortunately, even well-established methods like histochemistry,
magnetic resonance imaging (MRI), X-ray tomography, or
positron emission tomography (PET), are not really suitable for
regular intra-operative use. However, there is a clear need for an
intra-operative diagnostics especially in brain surgery. Vibrational
spectroscopy techniques (both Raman and infrared) complement
the standard methods for tissue diagnostics. [2] This contribution
shall review the current status of applicability of vibrational
spectroscopy under intra-operative conditions.
Both Infrared and Raman spectroscopy have the potential
for intra-operative use, because they can provide a biochemically
based profile of tissue in real time and without requiring additional
contrast agents. [3] Such marker-free techniques are particularly
required, because they avoid any perturbation of the sample under
investigation. Infrared spectroscopy is being used to examine
freshly resected tissue ex vivo in the operating theater while the
surgery is going on. The main potential in this case is the
identification of the borderline between normal and tumor tissue.
Other promising approaches include Raman fiber techniques and
non-linear Raman methods. Raman fiber techniques can easily be
combined with commercial endoscopes. Of the non-linear Raman
methods, Coherent Anti-Stokes Raman Scattering (CARS) attracts
the greatest research interest.
references:
1. Reiner Salzer, Biomedical Imaging: Principles and
Applications, John Wiley & Sons 2012, ISBN
0470648473
2. Reiner Salzer, Heinz W. Siesler, Infrared and Raman
Spectroscopic Imaging, Wiley-VCH 2009, ISBN
352731993X
3. Allison Stelling, Reiner Salzer, Matthias Kirsch, Stephan
B. Sobottka, Kathrin Geiger, Edmund Koch, Gabriele
Schackert, Gerald Steiner; Anal Bioanal Chem 400 (2011)
2745–2753.
Keywords: Vibrational spectroscopy; IR spectroscopy; Raman
spectroscopy; CARS (Coherent Anti-Stokes Raman Scattering);
AUGUst 26–30, 2012, PrAGUE, cZEcH rEPUbLIc