Chinese Journal of Chemistry, 2009, 27, 513—517
Full Paper
* E-mail: lihuaixiang@sdnu.edu.cn; Tel.: 0086-0531-86271517
Received June 26, 2008; revised November 6, 2008; accepted November 25, 2008.
Project supported by the National Natural Science Foundation of China (Nos. 60671010, 50534100, 20775045), and Natural Science Foundation of
Shandong Province (No. Y2006B29).
© 2009 SIOC, CAS, Shanghai, & WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Fluorescence Quenching of Pheophytin-a by Copper(II)
Ions
HU, Mingbo
a
(胡明波) LI, Huaixiang*
,a
(李怀祥) CHEN, Lusheng
a
(陈鲁生)
ZHANG, Huaibin
a
(张怀斌) DONG, Chuan
b
(董川)
a
College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan, Shan-
dong 250014, China
b
College of Chemistry and Chemical Engineering, Shanxi University, Taiyuan, Shanxi 030006, China
A method was developed for determination of Cu(II) ions quantitatively by measuring fluorescent intensity of
pheophytin-a (Pheoa) solution. The Pheoa was obtained by de-intercalation of magnesium from the porphyrin ring
of chlorophyll-a (Chla) extracted from fresh spinach leaves. Its two UV-Vis absorption peaks at 505 and 535 nm in
acetone solution have been observed but disappeared when the acetone solution of Pheoa was mixed with a Cu(II)
ion aqueous solution. A fluorescence quenching phenomenon was thus observed when the acetone solution of
Pheoa was mixed with an aqueous solution of Cu(II) ions. However, other physiologically relevant cations rarely
caused any quenching fluorescence of Pheoa under the same experimental conditions. Kinetics of the fluorescence
fading process was investigated by measuring the effects of Cu(II) ion concentration, reaction time and reaction
temperature on the fluorescence intensity of the Pheoa acetone solution. An activation energy of (10±1) kJ•mol
-
1
was estimated from Arrhenius empirical relation assuming that the interaction between the Pheoa and the Cu(II)
ions was the first order reaction. The calibration graph obtained with the fluorescence was linear over the Cu(II)
concentration range of 8.0×10
-
5
—8.0×10
-
7
mol•dm
-
3
with a detection limit of 8.0×10
-
7
mol•dm
-
3
for Cu(II)
ion.
Keywords fluorescence quenching, chlorophyll-a, pheophytin-a, Cu(II) ion, activation energy
Introduction
The chemical sensor has been mainly driven by the
requirements of higher and higher sensitivity, stability
and selectivity values.
1
Actually a sensor output can be
considered as the result of a process of synthesis of the
whole features of the environment, to which the sensor
is exposed.
2
One of the signal transduction ways is ob-
tained by monitoring fluorescence. There are various
advantages in using fluorescence.
3
Firstly, it is a pow-
erful visual phenomenon. Secondly, its sensitivity is
several orders of magnitude higher than that of absorp-
tion spectroscopy. Thirdly, a spectrofluorimeter is a
piece of equipment commonly available in many scien-
tific laboratories.
The chemistry of metalloporphyrins has been exten-
sively studied for many years in relation to the naturally
occurring tetra-pyrrole macro-cyclic compounds in
various biological systems, such as electron transfer
catalysts of heme-containing enzymes, oxygen-carrying
heme proteins, and light-harvesting pigments in photo-
synthetic systems.
4
Recently, synthetic porphyrins have
attracted attention in relation to the chemical and bio-
logical recognition.
5
A large number of porphyrin re-
ceptors have been designed for the detection of biologi-
cally and environmentally important small species such
as simple inorganic cations or anions.
6
Host-guest
chemistry using porphyrin scaffolds is unique owing to
several features of the porphyrin framework.
7
Copper is an essential trace element for both plants
and animals, including human beings but overdose
copper may be highly toxic to organisms. Therefore,
copper levels should continuously be controlled in
blood or drinking water.
8
It is known that among the most relevant transition
metal ions, Cu(II) is a paramagnetic ion in nature, which
quenches fluorescence emission of almost all fluoro-
phores around via electron or energy transfer mecha-
nism.
9
On the basis of fluorescence quenching, a num-
ber of fluorescence molecular sensors of Cu(II) have
been developed during the last several years.
10-12
How-
ever, no previously published papers have mentioned
about Cu(II) determination using natural porphyrins.
Natural porphyrin compounds, such as chlorophyll
a or b, are good fluorescent luminophors but have rarely
been used for fluorescent chemical sensors. Chloro-
phylls are aromatic compounds comprising a large
20-membered aromatic ring
13
and can act as ligands,
coordinating to a wide variety of metal ions via four
nitrogen atoms. Figure 1 shows the chemical structure
of chlorophyll a (Chla) and pheophytin a (Pheoa).
13