Zborník príspevkov z vedeckej konferencie - Department of ...

a) b)
2 2
1 1
3 3
3 3
Fig. 1 The excitation/emission matrices (EEMs) of diluted urine of (a) control and (b) experimental mouse; 1, 2, 3 - 1 st , 2 nd
and 3 rd fluorescence emission maximum; dilution 1:30, excitation wavelengths: 250 – 550 nm, emission wavelengths: 250 –
650 nm.
1
Fig. 2 Emission fluorescence spectrum of diluted urine of control and experimental mouse, (a) excitation 290 nm, (b)
excitation 370 nm; 1, 2, 3 - 1 st , 2 nd and 3 rd fluorescence emission maximum; dilution 1:30.
The concentration matrices of synchronous fluorescence spectra of urine were obtained for control and experimental
animals (Fig. 3). Synchronous spectral scanning revealed fluorescence (excitation/emission) bands at 363/383 nm and
484/504 nm for both control and experimental mice (Fig 4a). The synchronous fluorescence peak 363/383 nm corresponds to
the second emission fluorescence peak (330/416 nm) probably caused by 4-pyridoxic acid (317/420 nm) presence and the
synchronous fluorescence peak 484/504 nm belongs likely to the third emission fluorescence peak (450/520 nm) which may
by caused by flavins (450/525 nm) and its metabolites (Tab. 1).
The synchronous fluorescence spectrum of control urine exhibited two other fluorescence peaks, namely at 280/300 nm
by dilution 1:32 and at 393/413 nm by dilution 1:128, presumably due to suppressed concentration quenching of fluorophores
and reduced inner filter effect [3] (Fig. 4). The peak at 280/300 nm probably comes from proteins and the other peak 383/413
nm comes from NADH (340/460 nm) and from pterins (350/450 nm) (Tab. 1).
The position of fluorescence band at 363 nm is moved to short wavelength with dilution. This shift probably comes
from indol metabolites fluorescence (290/380 nm) and belongs to the first fluorescence peak (290/390 nm).
Implantation of human ovarian cancer cells SKOV-3 into the experimental mouse caused changes in the spectral
appearance: The intensity of fluorescence at 363/383 nm and 484/504, increased when compared to control (Fig4a).
Concomitantly, the new fluorescence peaks at 280/300 nm and 303/323 nm by dilution 1:128 have appeared (Fig. 4b) in
experimental mouse urine. These fluorescence bands can be preliminary attributed to the fluorescence of proteins.
Zborník príspevkov
z 18. medzinárodnej vedeckej konferencie
"Analytické metódy a zdravie loveka", ISBN 978-80-969435-7-9
- 89 -
2
3
hotel Falkensteiner, Bratislava
11. - 14. 10. 2010