Sensors and Actuators B 228 (2016) 774–781

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Sensors and Actuators B: Chemical

journal homepage: www.elsevier.com/locate/snb

Electrochemical determination of histamine in fish sauce using

heterogeneous carbon electrodes modified with rhenium(IV) oxide
Albana Veseli a,∗ , Majlinda Vasjari b , Tahir Arbneshi a , Ahmet Hajrizi c , L’ubomír Švorc d ,
Anchalee Samphao e , Kurt Kalcher c
a
Department of Chemistry, Faculty of Natural and Mathematical Science, University of Prishtina ‘Hasan Prishtina’ Nëna Terezë, 10000 Prishtina, Republic of
Kosovo
b
Department of Chemistry, Faculty of Natural Sciences, University of Tirana, Albania
c
Institute of Chemistry, Analytical Chemistry, Karl-Franzens University of Graz, Universitätsplatz 1, 8010 Graz, Austria
d
Institute of Analytical Chemistry, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Radlinského 9, Bratislava
SK-812 37, Slovak Republic
e
Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ubon Ratchathani University, Ubon Ratchathani 34190,
Thailand

a r t i c l e i n f o a b s t r a c t

Article history: The electro-catalyzed oxidation of histamine on heterogeneous carbon (carbon paste and screen-printed
Received 24 August 2015 carbon) electrodes modified with rhenium (IV) oxide was investigated for the purpose of its flow-injection
Received in revised form 16 January 2016 amperometric determination with a detection potential of −100 mV. Histamine was oxidized by the
Accepted 19 January 2016
mediator, which in turn was re-oxidized electrochemically. The developed histamine sensor showed a
Available online 22 January 2016
linear dynamic range up to 10 mg/L with a detection limit (3␴) of 0.2 mg/L (E = − 0.1 V) and 0.3 mg/L (E = −
0.15 V), respectively. The repeatability was 5.7% relative standard deviation (n = 10 measurements), the
Keywords:
reproducibility 14% (n = 5 sensors). The proposed sensor was applied for the assessment of the histamine
Histamine
Rhenium dioxide
content in fish sauce with the results in good accordance with spectrophotometry as a comparative
Flow injection analysis method.
Amperometric detection © 2016 Elsevier B.V. All rights reserved.

1. Introduction (Scombridae, Clupeidae, Engraulidae, Coryfenidae, Pomatomidae,

Histamine (2-aminoethyl-imidazole) is one of the biogenic
amines, which is produced by the enzymatic decarboxylation of the
corresponding amino acid histidine. It is also a byproduct of bac-
terial action during food processing and storage and hence could
be present in substantial amounts in fermented food stuffs and
beverages [1,2]
A high level of histamine correlates strongly with the num-
ber of microorganisms present in histidine rich foods (vegetables,
fermented foods and certain fish species). Amounts lower than
10 mg/kg of histamine indicate fish of good quality, a level of
30 mg/kg suggests significant deterioration, and a content of
50 mg/kg or higher is an evidence of definite decomposition. Safety
requirements on the histamine presence in seafood are regulated
by the European Commission [3]. For fish products, especially
from fish species associated with a high amount of histidine
∗ Corresponding author.
E-mail address: albana.veseli@uni-pr.edu (A. Veseli).
Scomberesocidae), the mean level must not exceed 100 mg/kg, and
no sample may contain a level exceeding 200 mg/kg [4,5].
Low levels of histamine in food are not considered as a serious
health risk, but when consumed in excess histamine may cause
specific undesirable physiological and toxic effects. The histamine
content in food is commonly used as an indicator of food deterio-
ration, and frequently regarded as one of the biomarkers in quality
control monitoring during food production, storage and transporta-
tion.
Both histamine deficiency and excessive histamine may cause
neurological and physical disorders in humans. One possible cause
for histamine deficiency is the presence of excess copper, as it
decreases blood histamine. Histamine ingested from food at a level
of more than 2.7 mg/kg body weight induces symptoms of his-
tamine intolerance. Adverse allergic reactions are experienced by
many sensitive individuals on ingestion of lower amounts already.
Histamine at levels usually exceeding 1000 mg/kg has been
implicated with certain food intoxications such as the cheese syn-
drome [6,7].

http://dx.doi.org/10.1016/j.snb.2016.01.085
0925-4005/© 2016 Elsevier B.V. All rights reserved.
 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781 775

Histamine has been determined after derivatisation with flu-

orescent reagents followed by chromatographic separation (e.g.,
HPLC in most cases). As the HPLC analysis of biogenic amines is
labor-intensive with regard to sample clean-up prior to the analy-
sis and requires trained personnel, other analytical methods have
been described such as capillary electrophoresis, immunochemical
methods such as ELISA, and even gas or thin-layer chromatography
[8,9]. To reduce the time needed for analysis and to offer a rapid
screening method for industrial food quality testing, some enzy-
matic methods and several enzyme sensors have been described so
far [10–13]. Chemical sensor applications, in general, and enzyme
biosensor applications in particular exhibit various advantages
such as allowing a more rapid analysis with less sample treat-
ment being required [14,15]. Modified electrodes are appreciated
for their main advantages, such as reduction of the analyte over-
potential (e.g., of H2 O2 ) and hence diminishing the interference
from other species by promoting electron transfer reactions which
may increase the selectivity, specificity and reproducibility of the
electrode surface, and improving the detection limit [16,17].
In the present paper, a sensor based on screen-printed car-
bon electrodes modified with ReO2 as a catalytic mediator for the
amperometric detection of histamine in flow-injection analysis
(FIA) is introduced. The optimization of the experimental condi-
tions and the application of the the developed sensor for assessing
the content of histamine in fish sauce is described.

2. Experimental

2.1. Chemicals and reagents

Biogenic amines (histamine, putrescine, cadaverine, tyramine,

tryptamine and serotonine) were purchased from Sigma–Aldrich
(Austria). All other reagents were of analytical reagent grade and
used directly without purification. Phosphate buffer solution (PBS,
0.1 M) was prepared by mixing aqueous solutions (0.1 M) of dis-
odium hydrogen phosphate and sodium dihydrogen phosphate to
the desired pH value. Stock solutions of amines were prepared by
dissolving suitable amounts of solid standards in PBS; they were
stored in the refrigerator at 4–6 ◦ C when not in use. Working solu-
tions of lower concentrations were freshly prepared on the day of
experiment by diluting the corresponding standard solutions.
p-Phenyldiazonium sulfonate used for photometric measure-
ments was prepared by mixing 1.5 mL 0.9% (m:v) of a sulfanilic
acid chilled solution and hydrochloric acid (4% m:m) with an aque-
ous sodium nitrite solution (1.5 mL, 5% m:v) in a volumetric flask
(50 mL) keeping it in an ice bath for 5 min. After that an additional
volume of 6 mL of the sodium nitrite solution was added and left
again under cooling for 5 further minutes before filling up to 50 mL
with chilled distilled water. The reagent was ready for use after
15 min of storage in the ice bath and was stable for 12 h [18].
Deionized water with a resistivity not less than 18 M–cm pre-
pared by a cartridge system (Milli-Q, Elix 3) was used throughout.

2.2. Instrumentation

The batch voltammetric system consisted of a potentiostat

(PalmSens, The Netherlands) connected to a laptop computer using
the corresponding software for data analysis (PSTrace). Carbon
paste electrodes CPEs (modified and unmodified) were used as Fig. 1. Cyclic voltammograms of histamine at a CPE unmodified (a); modified with
working electrodes. An Ag/AgCl/3 M KCl served as the reference Re2O3 (b); ReO2 (c); ReO3 (d);carrier phosphate buffer solution 0.1 M, pH 7.5; scan
and a platinum wire as the auxiliary electrode. All potentials cited rate 100 mV/s; solid curve-blank, dashed curve with 50 mg/L, dotted curve with
here are given against this reference electrode. 100 mg/L histamine.
All electrochemical flow injections experiments were per-
formed using a thin-layer electrochemical cell (CC-5, BASi), a
peristaltic pump (Pharmacia Fine Chemicals, Sweden), a sample
776 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781
Fig. 2. Possible model for the catalytic action of ReO2 on the oxidation of histamine; (A) catalytic redox cycle of rhenium (IV)-oxide via trivalent rhenium and the action of
histamine; (B) possible follow up reactions upon oxidation of histamine.
injection valve (5020 Rheodyne, Cotati, CA, USA), and an elec-
trochemical workstation (BAS 100B). The screen-printed working
electrode was fixed in the thin-layer cell via the Teflon gaskets
(thickness 0.19 mm) directly to the back plate of the cell with a
Teflon support as a holder. Silver conductive paint (Electrolube Ltd.,
Wargrave Berkshire, UK) was applied on one end of the SPCE, to
which a crocodile clamp was attached for electrical contact. The
software BAS 100 W, version 2, was employed for data processing.
For optical measurements a spectrophotometer (Cary 50, Var-
ian) was used.
Reference measurements were made with a UV–vis spectropho-
tometer (Cary 50Conc, Varian).
2.3. Preparation of working electrodes
Carbon paste electrodes: 1.00 g graphite powder (Ringsdorff
RWC, Bad Godesberg, Germany) and 360 ␮L paraffin oil were mixed
in an agate mortar by gently stirring with a pestle until uniformity
and compactness. For modification 50 mg of ReO2 were added per
gram of graphite powder.
Screen printed carbon electrodes (modified): 0.05 g of rhenium
(IV)-oxide was added to 1.00 g carbon ink (Electrodag 421 SS,
Acheson); the oxide-ink mixture was stirred for 10–30 min with a
glass-rod or stainless steel spatula and finally sonicated for 30 min
in an ultra-sonic bath (Transsonic 700/H, Elma® ). The resulting
mixture was immediately used for electrode fabrication using a
semi-automatic screen printer (SP-200, MPM, USA). The electrode
support was pre-etched sintered aluminum oxide (CoorsTek, UK).
The printed electrodes were dried at room temperature overnight
and then broken into individual sensor strips. The electrode was
inserted into the thin layer cell; electric contact was made with a
crocodile clamp.
2.4. Procedures
Cyclic voltammograms were recorded with carbon paste elec-
trodes either from −1.0 V to 1.0 V or from −0.2 to 0.2 V. The scan rate
was 100 mV/s, usually three cycles were recorded. Similar results
were obtained with slower scan rates (25 and 50 mV) with lower
currents. 100 mV were chosen for shorter analysis times. The sup-
porting electrolyte was phosphate buffer solution (0.1 M, pH 7.5),
the analyte concentration used for measurements was 50 mg/L of
histamine.
Flow injections analyses were done with screen printed car-
bon electrodes at a typical potential of −100 mV (or alternatively
−150 mV), the carrier solution was again 0.1 M phosphate buffer
solution (pH 7.5), flow rate of the pump was 0.6 mL/min and the
injection volume was 100 ␮L. The responses were evaluated using
the peak height.
All experiments were done at room temperature, 24.0 ± 1.5 ◦ C.
Spectrophotometric determinations of the analyte were per-
formed according to the reference method [18]. Thus, 5 mL of a
sodium carbonate solution (1.1%, m/m) were mixed slowly with
2 mL of the chilled reagent, i.e., p-phenyldiazonium sulfonate (as
described in 2.1). Then, 1 mL of the histamine standard or the
diluted fish sauce was added. The absorbance was measured at
496 nm. The concentration of histamine in the sample was obtained
from a calibration curve.
 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781 777

Fig. 3. Calibration curve for histamine within concentration 1–1000 mg L−1 at −100 mV. Supporting electrolyte phosphate buffer 0.1 M, pH 7.5; flow rate 0.6 mL/min; 100 ␮L
injection volume.

2.5. Food samples Table 1

Relative signals of interferences; 100% corresponds to a response generated by
100 mg/L histamine. Flow rate: 0.6 mL/min; operating potential: −100 mV vs.
In order to evaluate the applicability of the method to the deter- Ag/AgCl; carrier: phosphate buffer (pH 7.5; 0.1 M); injection volume: 100 ␮L.
mination of histamine in food samples, the analyte was quantified
C(␮g/L) Putrescine% Cadaverine% Tyramine% Tryptamine% Serotonine%
in various types of fish sauce. As samples three different brands
of fish sauces were used: SQUID, TIPAROS and NAM PLA OYSTER. 50a 1.9 6 17,7 24.5 25
Samples were analysed under optimal experimental conditions 100a 3,7 7.7 18,7 16 13
200a 5 8.2 20.6 12.8 11
by amperometric detection with ReO2 /SPCEs at −150 mV oper-
ating potential in FIA mode (Fig. 5). Though the baseline in FIA 50b −1.7 −2 −1 −5,5 −2,5
occasionally decreases in the negative current with subsequent 100b −3.9 −5.6 −3 −12 −10
200b −5.5 −7.3 −6.7 −14 −12.5
measurements, particularly when not waiting to its full recovery,
a
the signals measured as peak heights show a good reproducibility. (responses in the absence of histamine; separate solution method).
b
(responses in the presence of histamine).
Possible sample constituents such as aliphatic amines were tested
for their influence on the signal, but did not change the slope of the
calibration curve (see also interferences). using the calibration curves (Fig. 3) and optimized experimental
For the measurements in FIA and spectrophotometric method conditions. Standard solutions were prepared in the supporting
the diluted fish sauce was used without any other treatment such electrolyte. The contents of histamine in the samples ranged from
as filtered or centrifuged. Based on the histamine properties, con- 823 to 923 mg/L.
cretely his sensitivity in temperature, during the measurements The results obtained by the proposed sensor were compared
the samples were kept in ice bath. Fish sauce was diluted with with a photometric procedure [18]. p-Phenyldiazonium sulfonate
phosphate buffer solution (0.1 M, pH 7.5) prior to analysis to get a was reacted with histamine producing a colored compound. The
concentration in the linear range of the method. For FIA the dilution method was optimized and validated, and the best experimental
was done in ratio 1:100, for photometric determinations 1:200; conditions corresponded to the parameters given in the literature.
in the latter case at this dilution level the own absorption of the The differences between the both methods were assayed by the
sample was negligible. paired Student’s t-distribution test. At a confidence level of 95%
Thus, the baseline was stable for all injections, whereas it was there was no significant difference between the results of the two
slightly influenced by more concentrated samples. The results are methods [36]. Thus, it was concluded that the results obtained by
summarized in Table 3. The histamine content was calculated by two techniques were in good agreement, and based on this fact

Table 2
Comparison of the analytical parameters of the proposed method and other reported electrochemical methods for determination of histamine.

Electrode Method Linearity range (mg/L) LOD (mg/L) Samples Reference

Membrane with ␣-cyclodextrin and K tetrakis(p-chlorophenyl) borate Potentiometry 1–1000 0.5 Fish [1]
Boron-doped diamond Amperometry 5–900 4.5 − [31]
Gold Chronopotentiometry 2–100 0.27 Fermentedsausages [32]
Glassy carbon electrode Chronopotentiometry 2–90 1.31 Cheese [35]
Au ultramicroelectrode Cyclic voltammetry 0.005–5 0.00116 Tuna fish [34]
Glassy carbon electrode Chronopotentiometry 5–30 0.29 − [33]
Aluminum with a molecularly imprinted polymer Impedimetry not applicable 0.0016 bowel fluid [37]
SPCE with tetrathiafulvalene and histamine dehydrogenase Chronoamperometry 0.89–6.67 not mentioned octopus [38]
SPCE with ReO2 Amperometry 0.5–10 0.2 Fish sauce This work
778 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781

Fig. 4. Dependence of the peak height of histamine on the working potential in FIA with SPCEs modified with ReO2 as detector; (A) Io—baseline, Ip—peak maximum; (B) net
peak signals (Ip—Io); flow rate 0.6 mL/min, carrier 0.1 M phosphate buffer, pH 7.5, histamine solution 100 mg / L injection volume 200 ␮L.

the proposed method can be considered appropriate to be applied
for histamine determination in these kinds of samples. The appli-
cability of the sensor to other types of food is currently under
investigation.
The proposed method, using screen printed carbon electrodes
modified with ReO2 in FIA mode, is rapid because the sample
can be analyzed directly after proper dilution without any further
refinement steps. No additional cleaning processes of the electrode
surface are necessary. Very probably the method could be also
applied to other kinds of food, such as beer, fish, chicken meat and
fermented sausages which is currently the focus of further studies.
3. Results and discussions
3.1. Electrochemical characterization
Rhenium can exist in various oxidation states, which may form
corresponding oxides, which were studied as modifiers for car-
bon paste during this investigation, such as Re2 O3 and ReO2 , ReO3 .
Except Re(VII, VI, IV, III) also penta-, di- and monovalent species
may occur depending on the conditions [19,20]. In particular, if
complexing ligands are present, the probability and stability for
uncommon oxidation states become more realistic.
Fig. 1 summarizes cyclic voltammograms obtained in batch sys-
tem under the same conditions. All the curves do not contain
well-established unambiguously assignable redox signals in the
potential range of interest which indicates that the overall redox
behavior of the rhenium species is rather complex. The broad
signals, if occurring, suggest that sometimes probably more than
one distinct electrochemical reaction occurs involving complexed
and/or hydrated species.
Based on the voltammograms we may conclude that the species
ReO2 and Re2 O3 could be involved in the catalytic detection mech-
anism of histamine. ReO3 does not seem a very probable reaction
candidate because it exhibits the least significant changes in the
presence of the analyte.
For the detection of histamine in the negative potential range
two facts should be considered:
(i) a reduction current at the operating potential of −100 mV (or
below) can be observed for the ReO3 - and ReO2 -modified electrode
 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781
in the absence of the analyte which means, first, that ReO3 can be
excluded as a species to be directly involved in the mechanism,
because its formation from the modifier ReO2 would require an
oxidation. Second, another consequence is that ReO2 as a modifier
is reduced already at the operating potential, probably to Re2 O3 ;
(ii) histamine produces an oxidation signal, which suggests that
the analyte is oxidized and the mediator is reduced chemically;
consequently the reduced form of the mediator is reconstituted
again electrochemically causing an oxidation current.
In order to understand more exactly in which positions
or at which site of the histamine molecule the redox reac-
tion happens, different other organic amines (octylamine, 1,
5-diaminopentane, l-alanine, imidazole, 3, 4-dihydroxy phenylala-
nine adenine, thymine, xanthine, uric acid) were screened in flow
injection analysis. Besides histamine, only adenine and imidazole
yield significant current responses. Alkyl amines and their deriva-
tives produce only very small signals. Thus, it is reasonable to
conclude that the electrochemical conversion takes place at the
imidazole ring of histamine rather than at the amine group.
Due to the fact that histamine causes an oxidation signal, it may
be assumed that it is catalytically oxidized at the modified elec-
trode, because histamine itself is not electro-active at this potential
(−100 or −150 mV).
A possible overall reaction mechanism of the analyte and the
modifier at the electrode is illustrated in Fig. 2. scheme A [20,21–26]
taking into account all the facts which were mentioned above. Due
to the reductive background current at the operating potential a
reduction of the modifier can be assumed, probably to Re2 O3 . As
rhenium can form complexes in its various oxidation states it seems
probable that the first step of the catalytic conversion of histamine
is a complex formation with Re (III). We assume that this complex
is not stable but disintegrates into oxidation products of histamine
and a further reduced form of rhenium, probably Re (II). The lat-
ter is electrochemically oxidized to Re (III) yielding an oxidation
current in FIA. One possible oxidation pathway of the analyte is
its dimerization from primary radicals produced at the imidazole
ring. Another reaction could include the formation of an imida-
zolon derivative (Fig. 2, scheme B) [27–29]. As a conclusion Re2 O3
could be used as a modifier as well. In fact rhenium(III)-oxide shows
similar behavior as ReO2 but the signal is much smaller than with
the latter which could be explained by in situ formation of Re(III)
which shows higher catalytic activity (either directly or via a mixed
valence compound) than Re2 O3 .
3.2. Analytical parameters
The operation potential represents an essential parameter for
the performance of the sensor; it was optimized with FIA. The corre-
sponding hydrodynamic voltammograms within the range of −300
and 200 mV are displayed in Fig. 4.
The selected potential window seems proper for the application
of the sensor in biological matrices. They can be rather complex,
which means that they may contain a lot of oxidizable and reducible
substances. The higher the working potential, either negative or
positive, the higher is the risk of electrochemical conversion of
matrix constituents. Therefore, the aim of optimizing the operating
potential is to keep it as low as possible in both polarities, positive
and negative. In Fig. 2a the total currents (background, background
plus signal) are sketched; Fig. 2b shows the net signal response
which is of analytical relevance. In the range from −100 to 200 mV
we can observe only small signals. What is interesting is the fact
that the oxidation signal of the analyte increases when lowering the
operating potential to −300 mV. This in fact supports the reaction
mechanism which was suggested above. A more negative poten-
tial produces more of the reduced form of the modifier, i.e., Re2 O3 ,
which acts as an oxidant for the analyte. Thus, with more negative
779
potentials more oxidant is formed facilitating the oxidation of his-
tamine. Beyond −300 mV the signal levels off (not shown), because
other redox reactions take control of the overall electrochemical
process.
As most suitable potential −100 mV was chosen for different
reasons; first, the level of the background was much lower at
−100 mV than at more negative values yielding a high signal-to-
background ratio; second, the current signal was at an acceptable
level (it was only some four times higher at −200 mV); third,
the analysis time was significantly shorter than at more negative
potentials; and last, which is also important: the influence of inter-
ferences from matrix components is lower due to a less negative
potential. Taking all this in consideration most of the further studies
were done at −100 mV.
Calibration curves were established in order to check the sen-
sor performance at three appropriate potentials for the sensor;
−100, −150 and −200 mV in FIA mode using SPCEs modified with
ReO2 as the working electrode (Fig. 3). The detection limit (3␴)
was estimated from the standard deviation of FIA-peaks at 2.5 mg/L
histamine as about 0.2 mg/L. The repeatability of measurements
for 2.5 mg/L was about 4% RSD (n = 5 measurements), and the
reproducibility for 2.5 mg/L at different working electrodes (n = 5
electrodes) was 11%. There was no significant difference between
the different working potentials except the linearity range which
was better at −100 mV, 1–10 mg/L.
3.3. Interferences
The study of the principal interfering compounds is a crucial
step to evaluate the response of the sensor prior to the analysis
of samples to ensure its applicability. As potential sample a food
stuff was taken into consideration which contains a high amount of
biogenic amines due to the fermentation process, also obvious from
its smell, i.e. fish sauce. For this reason, the sensor response towards
some of the potentially interfering substances in fish sauce was
studied, namely, biogenic amines such as putrescine, cadaverine,
tyramine, tryptamine and serotonine.
The selectivity of the method was checked by monitoring the
sensor response of standard solutions of histamine in the absence
and in the presence of these five biogenic amines mentioned above.
Table 1 (values a) show the results for separate solutions, where
the signals of possible interferents at three different concentrations
(50, 100, 200 mg/L) are represented relatively to the standard sig-
nal of histamine (100 mg/L), which demonstrate that putrescine
and cadaverine produce only a negligible signal whereas tyramine,
tryptamine and serotonine cause some notable amperometric
response up to about 20% of the histamine peak current. Based on
their chemical structure aliphatic amines such as putrescine and
cadaverine could be considered as non-interfering agents giving
less than 10% of histamine signal up to 200 mg/L. Aromatic amines
exhibit some response which is surprisingly rather independent on
their concentration meaning that they have reached some satura-
tion at low concentrations already. All these amines yield reduction
currents in contrast to histamine showing oxidation signals.
The interferences were also measured in mixed solutions with
histamine (Table 1, values b). Three different concentrations of the
interferences were the same as above in the presence of 100 mg/L
histamine. At low concentrations the influence of all amines is
negligible, but increases at higher levels, especially in the case
of tryptamine and serotonine. In general, the histamine signal is
decreased.
The amount of interfering substances may vary in different types
of food samples, and in addition, it also evolves in time [30].
780 A. Veseli et al. / Sensors and Actuators B 228 (2016) 774–781
Table 3
Comparison of results for histamine concentrations, measured at three fish sauce
samples, using the new sensor in FIA mode and UV–vis method.
FIA (mg/L) UV–vis (mg/L) t calculated Confidence level (P = 95%)
923 ± 7 902 ± 9
860 ± 9 853 ± 1 2.7 3.182
823 ± 12 790 ± 2
3.4. Evaluation of the method
Although electrochemical methods are inexpensive and provide
high sensitivity, there are not many reports which reveal outstand-
ing performance of electrochemical methods for the determination
of histamine, probably due to the high oxidation potential (1.2 V vs.
SCE) of the analyte at which conventional electrodes such as glassy
carbon exhibit high background currents due to oxidation of the
solvent water or of the electrode material itself [33].
Anyway, electroanalytical methods offer the advantages such
as simplicity, precision, low cost of analysis and instrumentation
while keeping the consumption of solutions and reagents at a min-
imum. Using modified heterogeneous carbon electrodes for the
catalytically quantification of histamine is suitable for lowering the
potential required for histamine oxidation. Screen printed carbon
electrodes (rather than carbon paste) modified with ReO2 as cat-
alytic modifier yield mechanically more stable sensors than carbon
paste electrodes often combined with a longer shelf lifetime. The
screen-printed electrodes showed the same activity after half a year
of storage time.
In Table 2 analytical parameters of reported electroanalytical
methods for the quantification of histamine are summarized. Com-
pared to the other methods the approach presented here shows a
smaller dynamic range, but has a lower detection limit in many
cases. Nevertheless, there are a few methods with lower LOD
[34,37]. One is connected to microelectrodes in connection with
a complex cyclic voltammetric waveform with accumulations in
flow-injection mode combined with sample extraction as pretreat-
ment and mathematical corrections of the response curve [34],
another one to molecularly imprinted polymers with relative shifts
to a non-imprinted polymer and impedimetric measurements non-
linearly dependent on the analyte concentration. The detection
limit of the method presented here is sufficiently low enough to
detect histamine in the target sample matrix with good precision
simply after dilution of the sample. The proposed procedure repre-
sents a simple analytical method which does not require chemical
derivatization of the analyte and which can be used as a tool for
Fig. 5. A typical FIA amperogram for fish sauce; dilution factor (1:200) carrier phosphate buffer (0.1 M pH 7.5), operation potential −100 mV, flow rate 0.6 mL/min, injection
volume 100 ␮L.
routine analyses of certain food samples. Its applicability to the
determination of histamine in fish sauce is also shown here for the
first time.
4. Conclusions
The work presented in this paper demonstrates a possibility of
applying heterogeneous carbon electrodes modified with ReO2 for
the determination of histamine, an important biogenic amine.
Using ReO2 as a catalytic mediator has caused an increased per-
formance of the sensor for determining the analyte of interest by
reducing the overpotential of the analyte, thus diminishing the risk
of interferences from other substances, in particular from other bio-
genic amines.An LOD of 0.2 mg/L could be achieved with a dynamic
range up to 10 mg/L.
A very special outcome of this work was that the reaction
between histamine and ReO2 was different to other organic
amines (octylamine, 1, 5-diaminopentane, l-alanine, imidazole,
3, 4-dihydroxy phenylalanine adenine, thymine, xanthine, uric
acid), because the target analyte was oxidized negative operation
potentials of −100 and −150 mV whereas histamine itself is not
electro-active in the absence of the modifier, and alkyl amines
exhibit neglectible signals. It seems evident that the electrochemi-
cal conversion takes place at the imidazole ring of histamine rather
than at the amine group.
The proposed method, using screen printed carbon electrodes
modified with ReO2 in FIA mode, is rapid because the sample
can be applied directly after proper dilution without any further
refinement steps. No additional cleaning processes of the electrode
surface are necessary. The method was applied successfully to the
determination of histamine in fish sauce. Very probably the method
could be also applied to other kinds of food, such as beer, fish,
chicken meat and fermented sausages.
Acknowledgments
This work was supported by the Erasmus Mundus programme
of the European Commission in a partnership with the University of
Graz. The support of the bilateral program “JoinEU-SEE II” is grate-
fully acknowledged. This work was also supported by the Grant
Agency of the Slovak Republic (grant No. 1/0489/16).
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Biographies
Dr. Sc. Albana Veseli is the assistant of Analytical Chemistry at the Department of
Chemistry in University of Prishtina “Hasan Prishtina” in Kosovo since 2007. She
was graduated in 2005; defense her Master degree in 2009 and recently her PhD in
2014. She accomplished many research visits at the Institute of Chemistry, Analytical
Chemistry at the Karl – Franzens University of Graz, Austria as a Master and Ph.D.
student. She is a young researcher; her scientific interest covers the designing of
new sensors and biosensors based on heterogeneous carbon electrodes and their
applications in food and clinical analysis.
Prof. Dr. Majlinda Vasjari is the head of the Analytical Chemistry Group at the
Department of Chemistry, Faculty of Natural Sciences, in University of Tirana, Alba-
nia. She obtained a Ph.D. in Chemistry at University of Tirana (Albania) and followed
various postdoctoral researches in various international universities supported by
TEMPUS program, DAAD, Erasmus Mundus, etc. Her research is focused on envi-
ronmental monitoring and the development and application of electrochemical
sensors and biosensors. The materials used are inorganic particles and their com-
posites with biomolecules and biopolymers. She is author and co-author of around
50 publications.
Prof. Dr. Tahir Arbneshi has received his Ph.D. degree at the University of Tirana,
Albania.Currently, he is a Full Professor of Analytical Chemistry at the University of
Prishtina, Kosovo. He was a visiting professor during 2009–2010 at Jackson State
University, USA. Recently, his research focuses on gold nonmaterials for applica-
tions in materials science and biochemistry being author and co-author of about 10
publications.
Ahmet Hajrizi is currently a Ph.D. candidate in Analytical Chemistry at the Insti-
tute of Chemistry—Analytical Chemistry of the Karl—Franzens University of Graz,
Austria. He was graduated in Chemistry at the Department of Chemistry in Uni-
versity of Prishtina “Hasan Prishtina” in Kosovo. His recent research topic is the
electrochemical behavior of heterogeneous carbon electrodes modified with Tl2O3.
Assoc. Prof. Dr. L’ubomír Švorc is a researcher under the age of 35. He was grad-
uated in 2006, received his Ph.D. in 2009 and habilitated (Associate Professor) in
2015 for Analytical Chemistry at the Slovak University of Technology in Bratislava,
Slovak Republic. His interest covers the development and characterization of new
electrode materials for application of electrochemical sensors and biosensors based
on boron-doped diamond for solving the tasks of food, clinical and environmental
trace analysis. He is author and co-author of more than 70 publications.
Asst. Prof. Dr. Anchalee Samphao is a lecturer at Ubonratchathani University. She
was received her Ph.D. in 2007 and promoted to be Assistant Professor in 2013 in
Chemistry at Ubonratchathani University, Thailand. Her research is about develop-
ment of biosensors based on carbon paste electrode for clinical and environmental
analysis. She is author and co-author of more than 15 publications.
Ao. Univ. -Prof. Dr. Kurt Kalcher is the head of work group Electroanalysis and
Sensorics of the Institute of Chemistry—Analytical Chemistry of the Karl—Franzens
University of Graz, Austria. His main scientific interest lies in the development of
electrochemical sensors and biosensors based on carbon paste and other carbona-
ceous materials. Besides one of another focus is the development of simple analytical
devices including microprocessor control and data acquisition and handling. He has
authored–coauthored around 200 publications.