Monatsh Chem (2009) 140:11491154
DOI 10.1007/s00706-009-0167-x
ORIGINAL PAPER
Design and characteristics of tin (II) optode based
on immobilization of dithizone on a triacetylcellulose membrane
and its application in canned foods
Hossein Tavallali Solmaz Daneashian
Received: 7 January 2009 / Accepted: 25 July 2009 / Published online: 22 August 2009
Springer-Verlag 2009
Abstract The characterization of an optical sensor
membrane is described for the determination of tin (II)
based on the immobilization of dithizone on a triacetyl-
cellulose membrane. The membrane responds to tin (II)
ions by changing color reversibly from green to red in
buffer solution at pH 6 and wavelength 611 nm. This op-
tode has a linear range of 0.36.33 lg cm-3 (2.52
53.32 lmol dm-3) of Sn2? ions with a limit of detection of
0.15 lg cm-3 (1.26 lmol dm-3). The response time of the
optode was about 810 min depending on the concentra-
tion of Sn2? ions. The selectivity of the optode to tin (II)
ions at pH 6 was good. The sensor can be readily regen-
erated by exposure with EDTA solution. The color is fully
reversible, and the optical sensor could be fully regener-
ated. Experimental results showed that the optode could be
used as an effective tool in analyzing the tin content in
canned foods.
Keywords Optode
Sn2? ions
EDTA
Dithizone

Canned foods
Introduction
There is a huge variety of different chemical sensors, and
over the past 2 decades, the development and applications
of optical chemical sensors have grown rapidly. An optode
or optrode is an optical device that practically measures a
specific substance by absorbance, chemical bonding, or
physical entrapment [1]. The sensing element consists of
reagent dyes immobilized in organic or inorganic matrices.
H. Tavallali (&)
S. Daneashian
Chemistry Department, Payame Noor University, Shiraz, Iran
e-mail: tavallali@pnu.ac.ir; tavallali@yahoo.com
In most cases, a thin layer of polymer containing an
immobilized indicator dye is the core element of such
sensors [1]. Both the methods of immobilization and the
class of matrix exert a significant effect on the performance
of ion-selective layers [2]. Optochemical sensors based on
organic polymeric thin or thick films have been investi-
gated extensively in the last decade [3, 4]. Organic
indicator dyes and immobilization techniques play impor-
tant roles in the development and design of the optode.
Normally, the immobilization of the indicator is achieved
by absorption [5], chemical bonding [6], or physical
entrapment [7] into the support materials. Reaction with
the analyte changes the absorbance, fluorescence, reflec-
tance, and reflective index behavior of the sensitive layer
[811]. The development of optical sensors in analytical
chemistry is of great interest because of their possible
application in biology, biotechnology, and ecology [12,
13], and also because of their advantages such as small
size, freedom from electrical interference, low cost, safety,
the possibility of remote sensing, easy fabrication, good
sensitivity, and selectivity.
Tin is a widely sought metal and used in hundreds of
industrial processes, tin plates, pewter sheets, or jewelers
models, amalgam powder (in biological cases such as
dental filling) throughout the world. Interest in tin has
focused on its toxic potential in humans through the contact
of foods with tin-coated cans and tinfoil. Large amounts of
tin can accumulate in foods in contact with tin plates unless
these are lacquered or coated with resin [14, 15].
Tin can enter foods either from natural sources, envi-
ronmental pollution, packaging material, or pesticides.
Higher concentrations are found in processed food and
canned foods. Dissolution of the tinplate depends on the
food matrix, acidity, presence of oxidizing reagents
(anthocyanin, nitrate, iron, and copper), presence of air
123
1150 H. Tavallali, S. Daneashian

(oxygen) in the headspace, time, and storage temperature

[16]. According to the Food and Agriculture Organization
H
of the UN World Health Organization (FAD/WHO), the N N
maximum limit for tin in canned food is 250 mg kg-1. N N
H
Unfortunately, tin itself can slowly transfer into the food by S
many factors. It may cause stomach upsets, gastrointestinal
effects, and irritates the digestive tract of sensitive people Fig. 1 Structure of dithizone
when present in high concentration. When the concentra-
tion reaches a level higher than 250 mg kg-1 [17, 18], 1.6

organoleptic properties of the food can be observed, and (a)

there is some evidence suggesting that consumption of
food containing tin above 200 mg kg-1 has caused gas- 1.2

trointestinal effects [19].

There is a relationship between the concentration and

Abs.
0.8
the length of time after manufacture. After a can has been
opened, the amount of dissolved tin rapidly increases.
Fortunately, there are no long-term effects due to normal
0.4
intakes from tin in the food even if it is present in higher
levels. Several analytical methods have been developed for
the detection and determination of tin, including spectro-
0
photometry [20, 21], atomic absorption spectrometry [22, 400 450 500 550 600 650 700
23], atomic emission spectrometry [24], electrochemical /nm
[25], and spectrofluorimetric methods [26, 27]. 0.6
These methods have good selectivity, but they require (b)
well-controlled experimental conditions and careful sample 0.5
preparation. Although there are many reports on the
development of optodes for metal ion determinations, to 0.4
the best of our knowledge there has been no report on the
Abs.

use of an optical sensor for tin determination. 0.3

In this work, we describe the application of the metal
chromic reagent, dithizone, immobilized on a triacetylcel- 0.2
lulose membrane for determination of trace amounts of tin
(II) ions in juices and canned fish. The reaction takes place 0.1

within 810 min, and a color change occurs from green to

red at pH 6, which is spectroscopically detected in the 0
400 450 500 550 600 650 700
absorbance mode. /nm

Fig. 2 a Absorption spectra for a dithizone solution in the range of

Results and discussion 2.515 lmol dm-3 of tin at pH 6. b Absorption spectra of the optode
film in the presence of 110 lmol dm-3 tin at pH 6. Arrows indicate
the spectral trends in changing tin concentration from 2.5 to
Optical characteristics of the sensor membrane 15 lmol dm-3 in a and 110 lmol dm-3 in b

The optode membrane response to different metal ions was
investigated in universal buffer (0.04 M) at pH 6. The
membrane was first exposed to the buffer solution, and the
absorbance was measured at 611 nm. Then the sample
solution was added and the absorbance at 611 nm was
again measured after 10 min.
Dithizone (sym-diphenylthiocarbazone) contains azo
and hydrosulfide groups (S,N donating ligand), which can
form stable complexes with many metal ions under proper
conditions and is widely used in extraction spectropho-
tometry. The structure of dithizone is depicted in Fig. 1. If
the reagent is in excess and the aqueous phase is acidic
with pH 6, tin in the aqueous phase reacts with dithizone
dissolved in an organic solvent (ethylenediamine) to give a
highly colored red complex that distributes into the organic
phase. The absorption spectra of the membrane obtained
after being equilibrated in universal buffer solution (pH 6)
containing different concentrations of tin (II) ions are
shown in Fig. 2a. The spectral change (decrease in
absorption band at 611 nm and increase in absorption band
at 507 nm) is a result of the increase of tin ion concen-
tration in the membrane, which is due to the extraction of

123
Design and characteristics of tin (II) optode
tin ion into the membrane. In fact, the mechanism of the
response of optode is believed to be based on complexation
of dithizone and tin (II), which is accompanied by the
deprotonation of the ligand dithizone and release of proton
in the solution.
Spectral characteristics
Figure 2a and b shows the absorption spectra for free and
immobilized dithizone, which were obtained after being
equilibrated in buffer solutions (pH 6) containing different
concentrations of tin (II). The spectral changes are the
result of the addition of Sn2? ions and their reaction with
dithizone. The spectral characteristic of this sensor showed
two maxima at 611 and 507 nm. And those of the free dye
at 570 and 469 nm were observed. It is important to note
that the absorption spectrum of the immobilized indicator
is red shifted (or bathochromic shifted) in comparison to
that of the soluble form (611 instead of 570). This suggests
that the structured conformation of the immobilized indi-
cators is more planar than of its soluble analogue [28].
The effect of pH
The effect of pH on the complex was studied in the range
of 19 by changing the universal buffer from 1 to 9. A
maximum value in the sensor response was obtained at pH
6 and can be seen in Fig. 3. This pH was selected for
further studies.
Dynamic range
The absorbance measurements were expressed as the
absorbance difference, which is defined as the difference
between the absorbance of membrane with immobilized
0.03
0.02
Abs.
0.01
0 of Sn2?. Figure 5 shows the time dependance for the
0 1 2 3 4 5 6 7 8 9
pH
Fig. 3 Effect of pH on the optode film response
1151
0.5
0.4
0.3
Abs.
0.2
0.1
0
0 1 2 3 4 5 6 7 8
c (Sn 2+)/g cm-3
Fig. 4 The optode film response versus Sn2? ion concentrations in
the range 07 lg cm-3 (059 lmol dm-3) at pH 6
dithizone alone and the absorbance of the membrane in the
presence of tin (II) ions. Figure 4 shows the absorption
signals of the optode film to the various concentrations
of Sn2? ions in the range of 0.37 lg cm-3 (2.52
59 lmol dm-3) for tin (II) ions.
In this case, 6.5 lg cm-3 (54.73 lmol dm-3) was found
as the concentration of Sn2? ions that saturate the film. As
can be seen, the plot of the difference in absorbance versus
Sn2? ion concentration exhibited a linear range between
0.3 and 6.33 lg cm-3 (2.5253.32 lmol dm-3). The
detection limit that was estimated as the concentration of
analyte producing an analytical signal equal to three times
the standard deviation of the blank signal was found to be
0.15 lmol dm-3 for tin (II).
Response time
An important analytical feature of any optode is its
response time. The response time of the optode is con-
trolled by the time required for the analyte to diffuse from
the bulk of the solution to the membrane interface and to
associate with the indicator. The response time was tested
by recording the absorbance change from a pure buffer (pH
6) to a buffered tin solution of 8.42 lmol dm-3.
The membrane was found to reach 95% of the total
signal in about 810 min, depending on the concentration
10 absorption intensity of the membrane at 507 nm. In gen-
eral, the response time is more rapid when proceeding from
dilute to concentrated solutions.
123
1152
1.04
1.03
1.02
1.01
1.00
Abs.
0.99
0.98
0.97
0.96
0.95
0 200 400 600 800
t/s
Fig. 5 Typical response curve of the film optode at 507 nm as a
functional of time when the film was exposed to 8.42 lmol dm-3
Sn2? ions
Life time of optical sensor
The life time of membrane was determined by adding a
buffer solution (pH 6) in a cuvette including the film. The
signal was recorded at a wavelength of 611 nm over a
period of about 5 h. No significant loss of the indicator
occurs during this time. When the membrane was
exposed to light, no drift in signal was observed, and the
sensing phase was stable over the experiment with no
leaching of the indicator. However, prepared membranes
were kept under water when not in use to prevent them
from drying out. Additionally, the stability of response of
the film was investigated over 1 month under ambient
conditions, which indicated that the film was stable over
this period.
Reproducibility
The reproducibility and reversibility of the membrane in
the determination of tin (II) were evaluated by repeatedly
exposing the membrane to 8.42 lmol dm-3 tin solution.
The repeatability was evaluated by performing seven
determinations with the same standard solution of Sn2?.
The relative standard deviation (RSD) for the response of
one membrane towards a 8.42 lmol dm-3 of tin solution
was 1.9%. The reproducibility of the response of different
membranes was also studied. Six different membranes
were prepared from the same batch, and they were evalu-
ated by performing the determination of 8.42 lmol dm-3
tin. The RSD for the response between membranes was
3.2% for Sn2? ions. The results show the reproducibility to
be satisfactory.
123
H. Tavallali, S. Daneashian
Regeneration of the sensing phase
For a sensing phase to perform suitably, the color change
must be reversible. Some reagents, including HNO3, HCl,
SCN-, I-, S2O32-, sulfuric acid, histidine, 5-sulfosalicylic
acid, and EDTA, were studied as regenerating reagents.
The best result was obtained by applying EDTA, which
gave short membrane regeneration times (10 min). The on-
use durability of the sensor phase was obtained by subse-
quently placing the film in Sn2? ion and regenerating
solution. After regenerating seven times, a drift of about
2% was observed in the response of the sensor film.
Selectivity
To determine the selectivity of the optode film, the film
was tested for the determination of 8.42 lmol dm-3 Sn2?
ions in the presence of other metal ions, including Pb2?,
Cd2?, Cu2?, Hg2?, Ag?, Mg2?, Zn2?, Ni2?, Al3?, Co2?,
Bi3?, Fe2?, V4?, Na?, K?, and Ba2?. The tolerance limit
was taken as the concentration causing an error of 5% in
the determination of Sn2?. Samples containing a fixed
concentration of Sn2? ions, 8.42 lmol dm-3, and different
concentrations of other metal ions were analyzed by the
method. At the applied pH value no interference was
observed from even 100-fold excess of the above metal
ions except Co2?, Cu2?, and Ni2?. However, the interfer-
ence from these ions can be eliminated by using murexide
as the masking agent. Ions such as Cd2?, Hg2?, Ag?, Fe2?,
Al3?, and Zn2? were precipitated at this pH.
It is apparent from this study that discrimination among
different metals can be achieved by the control of several
parameters, including the absorption wavelength, the pH,
and the sample contact time in addition to the use of
masking agents.
Application
To evaluate the analytical applicability of the proposed
method, it was applied to determine tin in mineral water
samples and also in canned products. The water sample
was prepared by Kohrang mineral water factory, Shiraz,
Iran. The canned fish was prepared by the Sahele fac-
tory, Iran, and the fruit juice samples were prepared by
Shani factory, Iran. These samples were spiked with a
standard stock solution of tin (II) and determined by the
proposed optical membrane. The results are shown in
Table 1.
This proposed method also was applied to determine the
concentration of tin in different canned products (fruit juice
and fish). Prior to the determination, samples were digested
and diluted. The results are given in Table 2.
Design and characteristics of tin (II) optode 1153

Table 1 Recovery of tin from

Sample Tin (II) added Tin (II) founda Recovery (%)
spiked products before digestion
(mg kg-1) (mg kg-1)

Mineral water 1 0 n.d.b 96.0

1 0.96 0.03
Mineral water 2 0 n.d. 95.0
6 5.7 0.02
Mineral water 3 0 n.d. 98.0
20 19.6 0.03
Canned fish 1 125 127.7 4.1 102.2
Canned fish 2 150 152.3 4.5 101.5
Canned fish 3 175 176.4 7.5 100.8
Pineapple juice 1 25 25.2 1.9 100.8
Pineapple juice 2 50 49.4 2.1 98.8
Pineapple juice 3 75 73.9 1.8 98.4
Peach juice 1 40 38.7 1.1 96.7
Peach juice 2 60 61.4 1.4 102.3
Peach juice 3 80 79.7 1.2 99.6
Orange juice 1 25 24.6 1.9 98.3
a
Average of three Orange juice 2 50 50.2 1.7 100.4
determinations found by the
Orange juice 3 75 73.9 1.3 98.5
proposed method; bnot detected

Table 2 The total tin content of canned products (after digestion) reversible, has a low detection limit, and can be applied to
Type of sample Content of tina (mg kg-1)
the monitoring of tin in real samples.

Canned fishb 348 1.7

c
Orange juice 80 1.2
Peach juiced 100 1.10
Pineapple juicee 65 1.8
a b
Averages of three determinations standard deviation; after
8 months after production; cafter 6 months after production; dafter
9 months after production; eafter 7 months after production
Conclusion
The ion-selective optode is one of the optical chemical
sensors that have attracted interest since the 1970s [1].
Generally, the essence of the usefulness of the ion-
selective optode is the ion selectivity associated with the
highly selective complexation of an ionophore (either
neutral or charged) with an analyte ion in optical chem-
ical sensing.
In this work, a simple and non-extractive optode method
was developed for determination of trace amounts of tin
based on complex formation with a constructed membrane.
So far, there has been no report on the use of optical sen-
sors for tin (II) determination. The sensor responds to tin
ion by changing color reversibly from green to red. This
sensor can be regenerated readily with a solution of EDTA,
and it has a long lifetime. The response of the sensing
phase was reproducible, and the method is sensitive, fully
Experimental
Reagents
All reagents were the best available analytical reagent
grade, and all solutions were prepared with double distilled
water. The indicators dithizone (sym-diphenylthiocarbaz-
one), ethylenediamine, and EDTA solution were supplied
from Merck.
A stock solution of 1,000 lg dm-3 tin (II) was prepared
daily by desolving 0.095 g SnCl2
2H2O (Fluka) in double-
distilled water and diluting to the mark in a 50-cm3 volu-
metric flask. Standard solutions of lower concentrations
were prepared by suitable dilution of the stock solution
with distilled water. Buffer solutions were prepared from
boric acid/phosphoric acid/acetic acid (0.04 mol dm-3
each), and the pH adjustments were made by the addition
of sodium hydroxide solution (1 mol dm-3).
Instrumentation
A Perkin Elmer lambda 2 UV-Vis spectrophotometer was
used for recording the visible spectra and absorbance
measurements. The sensing membrane was placed and
fixed in a disposable glass cuvette, and all measurements
were performed in a batch mode. A Metrohm 3510 pH

123
1154
meter (model jenway) with a Metrohm double junction
glass electrode was used for pH adjustment.
Preparation of the sensor membrane
The immobilized indicator on the triacetylcellulose mem-
brane was prepared according to the following procedure:
The transparent triacetylcellulose membranes were pro-
duced from waste photographic film tapes that previously
had been treated with commercial sodium hypochlorite for
several seconds in order to remove colored gelatinous
layers. The films were treated with a clear solution of
dithizone (0.02 g) in 20 cm3 ethylenediamine for 7 min at
ambient temperature. Then they were washed with water
for removing excess ethylenediamine and loosely trapped
indicator. The membranes were finally washed with
detergent solutions and rinsed with water. Prepared mem-
branes were kept under water when not in use.
Digestion procedure for juice samples
To test the practical application of the present sensor, it
was applied to the determination of tin in fruit juice sam-
ples. Sample juice (5 cm3) was transferred into a 250-cm3
Erlenmeyer flask, and 10 cm3 concentrated sulfuric acid
was added. The solution was diluted to about 75 cm3 by
distilled water. Afterwards, it was cooled, filtered, washed
with water, and the filtrate was collected in a 10-cm3 cal-
ibration flask and diluted to the volume with water. For the
analysis, 0.5 cm3 of the obtained solution was transferred
into a 25-cm3 calibrated flask, and 0.02 g aluminum metal
was added in order to reduce Sn (IV) to Sn (II), and if
necessary, diluted to a suitable volume.
Digestion procedure for fish
The fish (2.0 g) was dried and ashed at 700 C. After that
the ash was dissolved with 10 cm3 of concentrated HCl and
diluted to 50 cm3 with distilled water and 1 mol dm-3
NaOH, so the pH 6 was reached. For the analysis, 0.5 cm3
of the solution was transferred into a 25-cm3 calibrated
flask, and 0.02 g aluminum metal was added in order to
reduce Sn (IV) to Sn (II), and if necessary, diluted to a
suitable volume.
123
H. Tavallali, S. Daneashian
Acknowledgment The authors wish to acknowledge the support of
this work by Payame Noor University Research Council.
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