Analytica Chimica Acta 558 (2006) 326–331

A new method of histamine colorimetry using

2,3-naphthalenedicarboxaldehyde on a
silica–gel column cartridge
Shigeyuki Oguri ∗ , Aki Mizusawa, Maiko Kamada, Minako Kohori
Laboratory of Food Hygiene, Department of Home Economics, Aichi-Gakusen University, 28 Kamikawanari,
Hegoshi-cho, Okazaki City 444-8520, Japan
Received 28 June 2005; received in revised form 29 September 2005; accepted 3 November 2005
Available online 19 December 2005

Abstract
In our work, we found, by chance, that 2,3-naphthalenedicarboxaldehyde (NDA) acts as a color development reagent to histamine (HA) in the
absence of a reductant, e.g., cyanide anion, even in a weakly acidic environment. The NDA-induced color development reaction to HA shows the
maximum absorption of 552 nm at pH 6 after a minimal interval of 10 min. By using this chemical property, a simple and convenient new method
of visual colorimetry of HA on a short silica–gel column cartridge (called an “HA cartridge” in this study, which consisted of 50 mg of silica–gel
packed into a 1.0 ml disposable syringe) was developed. The new visual method involves extraction of a 5 g sample with 5% trichloroacetic acid
solution (TCA, 35 ml), followed by neutralizing with 1 M sodium hydroxide. After loading the TCA extract (500 ␮l) into the HA cartridge, the
cartridge was then washed with a 0.1 M phosphate buffer, pH 6 (200 ␮l) and water (200 ␮l × 3 times). When a solution of 1 mM NDA in acetonitrile
(200 ␮l) was passed through the HA cartridge, the color inside the cartridge was observed to changed at 3 min after the NDA loading from white
to indigo-blue, for concentrations of HA ranging from 25 to 1000 mg kg−1 . This sequence requires only 5 min to perform.
© 2005 Elsevier B.V. All rights reserved.

Keywords: Allergy-like food poisoning; Colorimetry; Histamine; Histamine poisoning; 2,3-Naphthalenedicarboxaldehyde

1. Introduction
It is a known fact that many people suffer from allergic reac-
tions to various chemicals, house dust, pollen, animal hair, foods
of certain types, etc. Some components in food can cause either
food allergy or food intolerance, including allergy-like food poi-
soning (or histamine (HA) poisoning). Because both cases often
display the same symptom due to the body’s reaction to HA,
there are people who do not know the true etiology of their own
symptoms. To make it possible for people to determine the cause
of their symptoms and to prevent further cases of poisoning, and
because most cases of allergy-like food poisoning are caused by
ingested HA, a simple and reliable method for assaying for the
presence of HA in food is essential.
A fluorometric method officially approved by the Associa-
tion of Official Analytical Chemists (AOAC) (977.13) [1] has
∗ Corresponding author. Tel.: +81 564 34 1212; fax: +81 564 34 1270.
E-mail address: s-oguri@gakusen.ac.jp (S. Oguri).
been used routinely for 25 years as the basis for taking regulatory
action on fish containing HA. The method involves an extraction
with 75% methanol, removal of interfering compounds by an
ion-exchange column, derivatization with o-phthaldialdehyde
(OPA) [2,3], and measurement of fluorescence. Unfortunately,
this official method is difficult to perform and requires consid-
erable time and energy, hence its adoption can be viewed as
being somewhat problematical. To overcome these difficulties,
a wide variety of procedures for the determination of HA have
been published, such as detection by means of an oxygen-based
sensor electrode [4], or by an enzyme-based method [5,6], by an
enzyme-linked immunosorbant assay (ELISA) method [7], by
chemical colorimetry [8] and by chromatographic methods [9].
By using these techniques, recently, several commercially avail-
able test kits designed to determine HA in canned and raw fish
have become available. Although they are advertised as rapid,
easy to use, and capable of providing accurate results at low
cost, they all have similar merits and demerits [10]. For the past
decade, we have worked hard to develop simple and effective
methods for determining the presence of HA or other biogenic

0003-2670/$ – see front matter © 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2005.11.021
 S. Oguri et al. / Analytica Chimica Acta 558 (2006) 326–331
amines by employing OPA [11–16]. Regretfully, our methods
are still not optimally “user-friendly,” and are not, as yet, com-
mercially available. However, during our experiments regarding
2,3-naphthalenedicarboxaldehyde (NDA), which has a chemi-
cal structure similar to that of OPA, we found by chance that
NDA acts as a color development reagent to HA in the absence
of a reductant, even in a weakly acidic environment. By using
this chemical property, a simple and convenient new method of
visual colorimetry of HA on a short silica–gel column cartridge
(called an “HA cartridge” in this study) was developed.
2. Experimental
2.1. Reagents and solutions
NDA was obtained form Aldrich Chemical Company (WI,
USA). HA and other reagents were purchased from Wako Pure
Chemical Company (Osaka, Japan), and, being of the highest or
liquid chromatography (LC) grade commercially available, were
used without further purification. All aqueous solutions were
prepared using water purified with a Milli-Q purification system
(Millipore, Milford, MA, USA). HA or other amines, including
amino acids, were prepared by dissolving with water or with
buffered solutions to make a 0.05, 0.1, 0.5 and 1.0 mM solutions;
NDA solutions were prepared by dissolving with acetonitrile
to make 1.0 and 10 mM solutions. The 0.1 M buffer solutions,
having pH 2 and 4, pH 6, 7 and 8, and pH 9, were prepared
by dissolving sodium acetate, sodium dihydrogenphosphate and
sodium tetraborate decahydrate with water, respectively. Then,
each solution was adjusted to the desired pH with 1 M sodium
hydroxide. The 0.1 M phosphate buffer solution (pH 5) was used
for a capillary electrophoresis (CE) run buffer solution. The HA,
NDA, and other amine solutions were stored in a refrigerator
before usage.
2.2. Apparatus
Absorbance and fluorescence were measured with a Hitachi
Model U-2000 spectrometer and a Hitachi Model F-3010 spec-
trophotometer (Ibaragi, Japan), respectively, in a 3 ml volume
quartz cell. Food samples were homogenized with a Nissei
Model AM-3 universal homogenizer (Nihon Seiki Seisakusho
Company, Tokyo, Japan). For centrifugation of food samples,
a Hitachi Model Himac-CR 20 refrigerated centrifuge (Ibaragi,
Japan) was employed. For CE examination of the color reac-
tion mixture, the CE system consisted of a Jasco Model CE-800
(Jasco, Tokyo, Japan) equipped with an UV/Vis detector model
CE-971-UV (Jasco). All data were printed out by an intelli-
gent data processor a model 807-IT (Jasco). A capillary tube
of fused silica (25 cm effective length × 75 ␮m I.D.) was used
throughout the work. The window (0.5 cm) for detection was
made by removing the polyamide coating at a position located
15 cm from the cathodic end. Sample solutions were introduced
into the capillary tube from the anodic side by hydrostatic injec-
tion by raising the tube 5 cm higher than the level of the cathodic
electrode for 10 s. The electropherograms were recorded by
monitoring the absorbance at 210 nm. The applied voltage and
327
CE run buffer used throughout these studies was 6 kV and 0.1 M
phosphate buffer, pH 5, respectively.
For the liquid chromatography (LC) assay, the system con-
sisted of an intelligent PU-980 pump (Jasco, Tokyo, Japan)
used for isocratic-mode elution. Samples were loaded with a
Rheodyne syringe loading valve (Reodyne, CA, USA) with an
injection loop containing 20 ␮l of sample. The analytical column
was Wakosil-H5C18 (150 cm effective length × 4.6 mm I.D.;
Wako). The column was maintained at 40 ◦ C in a CO-965 col-
umn oven (Jasco). Column effluents were monitored at 220 nm
with a UV-970 UV/Vis detector (Jasco). All mobile phases were
degassed through a DG-980-51 on-line degasser (Jasco). The
flow-rate of the eluent was 1.0 ml min−1 . All chromatographic
data were also printed with the same intelligent data processor
as that for CE analysis.
2.3. Colorimetry for batchwise operation
Two millilitres of a buffer solution containing 0.1 mM HA or
other samples was mixed with 20 ␮l of a 10 mM NDA solution.
After keeping it at room temperature, the absorbance was read
at 552 nm. Note: the solution should be filtered with a dispos-
able syringe filter unit (Modele Dismic-13 cp, Adovantec Toyou,
Tokyo, Japan), if necessary.
2.4. CE examination
Twenty microlitres of 10 mM NDA solution was added to
2 ml of 0.1 mM HA in a 0.1 M phosphate buffer (pH 5.0) solu-
tion. Then, the mixture was examined with the CE system at
the following intervals, just shortly after starting the reaction,
15 and 30 min. The CE run buffer used here was matched to the
color development reaction solvent, 0.1 M phosphate buffer, pH
5.0.
2.5. Sample extraction
The extraction of HA from food samples was carried out as
follows: A 5 g amount of sample was homogenized with 40 ml of
5% trichloroacetic acid (TCA), then diluted to 50 ml as the final
volume with the same solvent as was used during homogeniza-
tion. Subsequently, the extract was transferred to a centrifuge
tube and centrifuged (11,200 × g) at 4 ◦ C for 10 min. The super-
natant was stored in a 50 ml-plastic bottle with a stopper in a
refrigerator until just before use. For LC examination, the sample
was extracted with methanol instead of TCA. After centrifug-
ing the extract the same way as above, 1.0 ml of the supernatant
was evaporated to dryness in vacuo. Subsequently, the resulting
residue was reconstituted with 10 ml of water prior to examina-
tion by LC.
2.6. Assay by LC
The LC method employed was one that was slightly modi-
fied from the ion-pared LC with UV detection method reported
by Yagi [17]. The mobile phase was prepared by dissolving 1-
octanesulfonic acid (432 mg) with a mixture of water (830 ml),
328 S. Oguri et al. / Analytica Chimica Acta 558 (2006) 326–331

acetonitrile (120 ml) and 0.2 M phosphate buffer (pH 3.0; 50 ml).
HA was eluted at 18.3 min when 10 ␮l of 0.01% HA standard
solution (this was equivalent to a 100 mg kg−1 HA-food sample)
was examined by LC.

2.7. Fabrication of HA cartridge

A short silica–gel column cartridge (or HA cartridge) was

fabricated as follows: (1) First, a small amount of cotton was
stuffed inside the bottom of a 5 ml disposable syringe (Terumo,
Osaka, Japan) to retain packing materials. (2) Fifty milligrams
of silica–gel (Silica–gel 60, Merck) was put on top of the cotton
inside the bottom of the syringe. (3) The silica–gel was “sand-
wiched” by inserting another small amount of cotton into the
silica–gel in the syringe. (4) Finally, the material inside of the
syringe was compacted by tamping with a thin rod.
Fig. 2. Picture showing the color developments of a 1 mM HA solution in 0.1 M
buffer having different pHs of 2, 4, 5, 6, 7, 8 and 9, after 30 min at room tem-
perature.
the presence of HA. Therefore, the following experiments were
carried out.
3.1. Effect of pH on color development

2.8. Assay with HA cartridge
One millilitres of TCA extract was mixed with a 250 ␮l of 1 M
sodium hydroxide. Then, the whole mixture was passed through
the HA cartridge; subsequently, the cartridge was washed with a
200 ␮l of 0.1 M phosphate buffer (pH 6), followed by a 1000 ␮l
of water. Finally, 200 ␮l of a 1.0 mM NDA solution was passed
through the HA cartridge slowly, and the color inside the HA
cartridge was observed after 3 min.
3. Results and discussion
As mentioned above, NDA [18,19] generally reacts with pri-
mary amino compounds in an alkali environment in the presence
of cyanide anion to yield a fluorescence derivative as shown
in Fig. 1. Although OPA can react with HA in the absence of
2ME [2,18,20–22], it was believed that the reaction of NDA and
HA did not occur when the cyanide ion or other reductant was
absent from the reaction mixture; similarly, it was believed that
the reaction did not occur in a non-alkaline environment even
when cyanide anion was present, because NDA was chemically
less active than OPA with regard to nucleophilic atoms present
in the HA molecule, such as nitrogen. However, when a NDA
solution was added to a HA solution in week acid buffer, we
found, by chance, that the mixture changed color from colorless
to indigo-blue. We realized that this meant that NDA could be
utilized as a color-development reagent for visually detecting
The specific NDA-induced color development reaction to HA
is shown in Fig. 2 under the different conditions of pH after start-
ing the reaction at room temperature. Under the condition of pH
greater than 6, both color development and precipitation were
observed in each solution. In addition, both pH 5 and 6-solutions
showed color development shortly after adding the NDA solu-
tion. Color tone changed only slightly as a variable with reaction
time, and some precipitation appeared only after an elapse of
several minutes or several tens of minutes after starting the reac-
tion; notably, the solutions were still colored and it was easy to
recognize the difference between the color of HA solution and
HA free solution even 6 h after the start of the reaction. As color
development appeared to depend both on elapsed time and the
pH of the solution, the effect of reaction time on color devel-
opment was evaluated first, using buffers having different pHs
of 3, 5, 6, 7, 8 and 9, respectively. Fig. 3 shows the effect on
NDA-induced absorbance with HA at different elapsed times of

Fig. 1. Schematic showing the chemical reaction of 2,3-naphthalenedicar- Fig. 3. Absorbance levels with change of time at different pHs of: (
) 3.0; ()
boxaldehyde (NDA) and a primary amino compound in two cases, i.e., with 5.0; () 6.0; () 7.0; () 8.0; (䊉) 9.0. The absorbance of each reaction solution
cyanide anions present and with cyanide anions absent, and with two conditions (2 ml of 0.1 mM HA in buffer and 100 ␮l of 10 mM NDA solution) was measured
of pH 10 or pH of less than 10. at 552 nm.
 S. Oguri et al. / Analytica Chimica Acta 558 (2006) 326–331
0.5, 1.0, 2.5, 5.0, 10, 20 and 30 min after starting the reaction.
Each trace was given by measuring the absorbance at 552 nm
(the reason why a wavelength of 552 nm was chosen will be
explained in Section 3.2). During these experiments, some pre-
cipitation appeared in the cases of pH 5 (at 40 min after starting
the reaction), pH 6 (after 15 min) and pH 7 (after 5 min); each
solution was, therefore, measured after filtering or centrifuging
to remove the precipitates. Thus, it was observed that the short-
est time to reach the maximum absorption of 552 nm occurred at
pH 6 after a minimal interval of 10 min. When the pH was either
less than pH 2 or more than pH 7, very little coloring reaction
was observed. In case of a solution having pH 5 or 6, it occurred
shortly after starting the reaction. In addition, with regards to
the reaction of NDA and HA under the condition of pH 5 or 6
at room temperature, no fluorescence was observed, neither at
1 min nor at 10 min after starting the reaction.
Next, to study the effect of reaction time on color develop-
ment, pH 5 was employed in order to prevent the undesirable
occurrence of precipitation during the experiments. Absorption
spectrums were measured at the following separate intervals, 0
(before adding NDA solution), 5, 10, 20 and 30 min, as shown
Fig. 4(A)–(E), respectively. At 5 min after starting the reaction
(Fig. 4(B)), the peak appeared at 552 nm, which suggested that
some product formation occurred in this reaction. The increase
of absorbance at 552 nm occurred rapidly until 5 min after the
start of the reaction, but increased very slowly at later times.
Thus, the wavelength of 552 nm was chosen as the indicator
Fig. 4. UV/Vis spectra of the reaction mixture of HA and NDA at different time
lapses: (A) 0 min; (B) 5 min; (C) 10 min; (D) 20 min; (E) 30 min. Each spectrum
was obtained by measuring the reaction solution (2 ml of 0.1 mM HA in a 0.1 M
buffer at pH 5.0 and 20 ␮l of a 10 mM NDA solution).
329
of the present coloring reaction in this study. The absorbance
peak at 262 nm, corresponding to the naphthalene ring of NDA,
decreased slowly with the reaction time and the broadband
absorbance appeared at around 245 nm (Fig. 4(D) and (E)) at
20 min after the start of the reaction. During this interval, the
change in color was observable with the naked eye.
To study the reaction further, the reaction mixture was exam-
ined with CE. Because the color reaction depended on pH, the
CE run buffer used here was matched to the color development
reaction buffer, 0.1 M phosphate buffer having pH 5, to avoid the
effect of pH on the reaction products during their CE migration.
Just shortly after starting the reaction, the two peaks at migration
times of 3.7 and 11.5 min were identified, which corresponded
to HA and NDA, respectively, as shown in Fig. 5(A). At 15 and
30 min after starting the reaction, more than six peaks corre-
sponding to reaction products (or by-products) were observed
between the both HA and NDA peaks on both electropherograms
(Fig. 5(A) and (B)). These results suggest that complex products
are yielded in this reaction. Regretfully, the chemical structures
corresponding to those six peaks have not yet been identified.
Regarding this chemical reaction mechanism, we suspect that it
is a type of reaction similar to that, which occurs with OPA in
the absence of a reductant [20–23].
Fig. 5. Electropherograms of the reaction mixture (2 ml of 0.1 mM HA in a 0.1 M
buffer at pH 5.0 and 20 ␮l of a 10 mM NDA solution) at different times: (A)
just after starting the reaction; (B) 15 min after starting the reaction; (C) 30 min
after starting the reaction, all cases at room temperature. Peaks indicated in (1)
and (2) on each electropherogram correspond to HA and NDA, respectively.
The arrows indicate the time at which CE analysis was started. (Refer to the CE
conditions described in Section 2.)
330 S. Oguri et al. / Analytica Chimica Acta 558 (2006) 326–331

3.2. Effect of HA concentration on the color

development

To make sure that this coloring reaction developed in pro-

portion with increasing amounts of HA, the effect of HA con-
centration on color development was evaluated by plotting the
absorbance (552 nm) at concentrations of 0.05–1.00 mM HA
in pH 6 after a 10 min-reaction with NDA. The absorbance
increased linearly with increasing concentrations of HA. This
Fig. 6. Picture showing the color development tests of fresh tuna-meat spiked
implied that colorization took place between HA and NDA. with HA at concentrations of 0, 25, 50, 100, 500, and 1000 mg kg−1 , just 3 min
In addition, the linearity and reproducibility (at 0.5 mM) were after an 1 mM NDA solution was passed through an HA cartridge.
calculated as r = 0.997 and 1.38% center of value (n = 5), respec-
tively.
3.4. Cross-reactivity tests
3.3. Detection of HA with HA cartridge
Cross-reactivity tests for the present coloring reaction were
separately performed by using both batchwise (in solution) and
By causing this coloring reaction to take place in an HA
HA cartridge methods. For these studies, 18 kinds of amino
cartridge, HA was detected in food. The HA cartridge can
acids, viz., alanine (Ala), arginine (Arg), aspartic acid (Asp), glu-
also serve as device for removing components other than HA
tamic acid (Glu), glycine (Gly), histidine (His), hydroxyproline
in food extracts that might react with NDA to develop col-
(Opr), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine
oration before developing the NDA–HA reaction on the HA
(Met), phenylalanine (Phe), proline (Pro), serine (Ser), taurine
cartridge. To determine the best way for detecting HA in food
(Tau), threonine (Thr), triptophan (Trp), valine (Val),) and six
samples with the HA cartridge, a number of experiments were
kinds of bioamines, viz., cadaverine (Cad), putrescine (Put),
carried out, such as what amount of the 1 M sodium hydrox-
serotonin (5HT), spermidine (Spr), tyramine (Tyr), and his-
ide should be added to a sample TCA extract, what amount of
tamine (HA) were employed. For the batch test, each amine
sample extract should be injected into the HA cartridge, which
solution was prepared with 0.1 M phosphate buffer, pH 6, and
kinds of buffer solution should be employed for washing the
the test was done by measuring absorption 5 min after starting
cartridge, and so on. Regarding the packing material for the
the reaction to avoid occurrence of the undesirable some pre-
HA cartridge, the performance of various materials was also
cipitation during the measurements. HA, His and 5HT showed
examined to determine optimum performance. After doing var-
absorbance of 1.55, 0.35 and 1.12, respectively. Trp was col-
ious combinations of washing solvents and packing materials,
ored yellow, but the absorbance at 552 nm showed 0.03. All
such as ion-exchange resin and octadecyl silane (ODS) parti-
other amines showed absorbance of less than 0.03. Regarding
cles for LC, it was concluded that the following 4-step protocol
the HA cartridge tests, each sample of tuna-meat spiked with
resulted in the best performance to date: (1) addition of 1 M
one of the amino acids or biogenic amines (0, 25, 50, 100, 500
sodium hydroxide to an sample extract, (2) passing the extract
and 1000 mg kg−1 ) was separately tested by the same method
through the HA cartridge, (3) washing the HA cartridge, and
as mentioned in Section 3.3, but none of the amines, except HA,
(4) loading a NDA solution. This sequence requires only 5 min
showed a change of color inside of an HA cartridge, even when
to perform.
the concentration was at the highest level (1000 mg kg−1 ). This
There are many kinds of pseudo-allergens containing HA,
suggested that, for the batch test, the four compounds, except
such as fruit, vegetables, wines, and certain kinds of animal
for HA showing a color change, could be removed by washing
meat and fish meat. Scombridae or Tuna is by far the riskiest
the HA cartridge with a buffer solution (pH 6) and water before
food regarding contamination with HA. Thus, raw tuna-meat
inciting the NDA–HA reaction on the HA cartridge.
was chosen as the test sample for this experiment; control sam-
ples spiked with HA to make concentrations of 0, 25, 50, 100,
500 and 1000 mg kg−1 were separately prepared for this study. 3.5. Testing of real samples
Before preparing the spiked samples, sample which contained
no HA (less than 1.0 mg kg−1 ) was checked with the LC method. To demonstrate real-sample applicability, samples of fresh
After treating the sample extracts the above-mentioned proce- tuna-meat were first aged for 8 and 24 h at room temperature
dures, the color change inside each HA cartridge was observed and then examined by means of the previously reported proce-
after 3 min, as shown in Fig. 6. Accordingly, the visual detec- dure [17] with a slight modification. The assay results showed
tion limits were observed at 25 mg kg−1 HA. Furthermore, these HA levels of 28 and 625 mg kg−1 in the 8 and 24 h-aged sam-
observations were possible even after an elapse of several hours, ples, respectively. Then, both of the above two samples were
although the tone of the observed color changed slightly with separately tested with the present method using HA-cartridge.
reaction time. It should be noted, however, that it was not pos- The resulting color tones of both cartridges are shown in Fig. 7.
sible to detect HA directly with these samples using batchwise To estimate the concentration of HA present, the color tone was
colorimetry, due to colorization of the HA-free sample extract compared with those of the standards shown in Fig. 6. The color
(0 mg kg−1 HA). tone of the cartridges of the 8 and 24 h-aged samples were
 S. Oguri et al. / Analytica Chimica Acta 558 (2006) 326–331
Fig. 7. Two samples of tuna-meat, aged for 8 h (A) and 24 h (B) at room tem-
perature, were tested. The LC assay results showed that HA contents in (A) and
(B) were 28 and 625 mg kg−1 , respectively.
somewhere between that of 25 and 50 mg kg−1 -spiked stan-
dard sample and 500 and 1000 mg kg−1 -spiked standard sample,
respectively. These results correspond closely with the results of
the LC assays. Although the present method cannot be used as an
alternative of the official method due to the poor detection sen-
sitivity and poor quantitative determination, we believe that the
method using HA cartridge, or so called “HA-detection kit,” pre-
sented here can be considered as a candidate for a first-screening
or pre-testing for the presence of HA in food. Based on our lim-
ited clinical information, allergy-like food poisoning caused by
HA requires that the food in question be contaminated by more
than 500 or 1000 mg kg−1 of HA. Taking this into account, the
present sensitivity (25 mg kg−1 ) and selectivity were sufficient
to “rule out” pseudo-allergen containing HA as a possible cause
of allergy-like food poisoning.
4. Conclusions
By using the new visual color development reaction of HA
with NDA, we developed a simple detection method for the pres-
ence of HA in a food sample. The present method using the HA
cartridge has the following merits: (1) the present HA kit has
a detection sensitivity 25 mg kg−1 , which might be enough to
prevent the occurrence of HA poisoning, by preventing the sale,
serving and ingesting of HA contaminated food which has been
“pre-screened” for the presence of HA; (2) the time needed to
measure HA in a TCA extract of food is only 5 min, which is as
short or shorter than any other method; (3) It is much easier to
331
preserve the HA kit in good condition than with the other meth-
ods, because the HA kit includes stable materials, not unstable
biological ones, such as enzymes or a HA specific antibodies;
(4) the HA kit is HA-selective.
Our final goal of HA analysis in food is to produce an inex-
pensive, handy, portable-type kit that can be used to confirm the
presence or absence of HA in food by anyone at anytime and
anywhere, without any special techniques. Although the present
method has less detection sensitivity in comparison with the
official AOAC method, the HA cartridge has sufficient sensitiv-
ity to help prevent allergy-like food poisoning induced by HA
contamination.
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