Food Chemistry 403 (2023) 134345

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Colorimetric determination of peroxide value in vegetable oils using a

paper based analytical device
Elham Ghohestani , Javad Tashkhourian *, Bahram Hemmateenejad *
Department of Chemistry, Shiraz University, Shiraz, Iran

A R T I C L E I N F O A B S T R A C T

Keywords: Peroxide value (PV) is one of the most typically used quality parameters to monitor lipid oxidation. Here, a
Peroxide value simple paper-based analytical device (PAD) has been developed to determine PV in vegetable oils. The analysis is
Colorimetric method based on setting up the iodometric titration, where hydroperoxides in the oil are reacted with excess iodide ions
Paper-based analytical device
to generate iodine molecules, on the paper substrate. The device is composed of two paper layers acting as re­
Vegetable oils
action and detection zones, aligned in a metallic mold. A well-defined inverse logarithmic calibration curve was
established between the measured PV by the official iodometric method (ISO;3960, 2017) and the color intensity
of PAD. It offered a working range of 0.01–30.0 meq/Kg. The limit of detection of 0.015 meq/Kg demonstrated
enough sensitivity of the method to estimate peroxide value in edible oils. On-site and visual detection, low cost,
simplicity, and less solvent consumption are advantages of the proposed device.

1. Introduction
The oxidation of lipids (fats and oils) during processing and storage
causes food toxicity (Mehta, Darji, & Aparnathi, 2015; Pilipenko,
Nilova, Malyutenkova, & Tverskoi, 2020). Heat and light accelerate the
degradation of hydroperoxides, as the primary product in the initial
stages of the oxidation, and produce dangerous secondary products such
as epoxides, alcohols, aldehydes, and ketones (Yu, Li, Sun, Dong, &
Wang, 2015; Zhang, Zuo, & Ye, 2015). Due to the toxicity of these
compounds, quantifying them is necessary to determine the degree of
oxidation of oils. Peroxide value (PV) is often used as an index to
monitor oil oxidation and oil quality control during processing, storage,
and marketing. PV is defined as milliequivalent or millimole of hydro­
peroxides per kilogram of oil (meq O2/kg of oil) (Mahboubifar, Youse­
finejad, Alizadeh, & Hemmateenejad, 2016; Yu et al., 2015; Yu,
Vandevoort, & Sedman, 2007).
Several analytical methods have been reported to measure the PV in
food. Based on the recommendation of the Association of Official Agri­
cultural Chemists (AOAC, 2000), International Organization for Stan­
dardization (ISO, 2017), and American Oil Chemists’ Society (AOCS,
2009), PV is measured based on the iodometric titration method. There
are other methods, including the FOX method (ferrous oxidation-xylenol
orange method) (Shantha & Decker, 1994), ferric thiocyanate method
(International Dairy Federation, IDF;74A, 1991) and International
standard method (ISO, 2006), which is based on the oxidation of iron.
Whilst the mentioned techniques are relatively simple, practical and
easy to perform, and are officially accepted methods (e.g., ISO, 2017;
AOAC, 2000; Aocs, 2009), they come with some drawbacks. For
example, titration methods generally use a large number of samples,
reagents, and solvents, which are all environmental concerns (Zhang
et al., 2021). Fourier-transform infrared (FTIR) spectroscopy has also
been used to determine the PV (Mahboubifar et al., 2016; Marina, 2014;
Meng, Ye, Nie, Pan, & Jiang, 2015; Rohman, Che Man, Ismail, &
Hashim, 2011; Yu et al., 2007). This method is sensitive, non-
destructive, and requires minimal sample preparation, but on the
other hand requires expensive equipment and well-trained personnel
(Zhang et al., 2021).
Recently, paper-based analytical devices (PADs) have attracted the
interest of scientists as simple and affordable tools for solving chemical
analysis problems. These devices have the advantages of easy operation,
simple fabrication, low cost, availability, and portability, which are very
promising for point-of-care analytical applications and on-site analysis,
where laboratory reagents, medical equipment, glassware, and in­
struments are lacking (Cai, Ouyang, Song, & Yang, 2020; Myers, Ker­
nisan, & Lieberman, 2015; Taghizadeh-Behbahani, Hemmateenejad, &
Shamsipur, 2018). Self-pumping capability, which is because of the
capillary action of paper, makes it an interesting platform for the
fabrication of microfluidic devices. Among different fabrication

* Corresponding authors at: Chemistry Department, Shiraz University Shiraz 71454, Iran.
E-mail addresses: tashkhourian@shirazu.ac.ir (J. Tashkhourian), hemmatb@shirazu.ac.ir (B. Hemmateenejad).

https://doi.org/10.1016/j.foodchem.2022.134345
Received 8 July 2022; Received in revised form 7 September 2022; Accepted 16 September 2022
Available online 20 September 2022
0308-8146/© 2022 Elsevier Ltd. All rights reserved.
E. Ghohestani et al.
Fig. 1. A schematic of procedure colorimetric detection using paper based analytical device for peroxide value determination.
strategies, three-dimensional microfluidic structures have gained im­
pressions since the fluids follow through the tiny depth of paper, which
is in the micrometer dimension. Various researches have been done so
far concerning the development and application of PADs for quality
monitoring and safety of food (Cinti, Basso, Moscone, & Arduini, 2017,
Cinti, Moscone, & Arduini, 2019; Mostafapour, Mohamadi Gharaghani,
& Hemmateenejad, 2021; Zhang et al., 2015). In our research group, we
have used 3-dimensional paper-based microfluidic devices for different
applications (Pesaran, Rafatmah, & Hemmateenejad, 2021; Sharifi,
Tashkhourian, & Hemmateenejad, 2020).
In the current study, we aimed to fabricate a PAD that is able to
quantify PV of edible and cold press oil samples at different storage
conditions. The chemical reagents that are used in iodometric titration
were preloaded onto two layers of paper, stacked on each other to
provide a 3-dimensional microfluidic device. To this end, two layers of
the PAD were aligned vertically in a cylindrical metallic mold to facili­
tate sample transport based on vertical flow. The device was calibrated
based on the official method (ISO, 2017).
2. Materials and methods
2.1. Chemicals and materials
Whatman filter paper grade 2 was purchased from Whatman GE
Healthcare (U.S.). Chloroform, starch, sodium thiosulfate, and acetic
acid were obtained from Merck Chemical Company. Potassium iodide
was bought from Chem-Lab Company.
Saturated-Potassium iodide solution (saturated-KI), was prepared by
dissolving excess KI in freshly boiled distilled water (DW). Fresh solu­
tions were prepared daily and stored in the dark. The starch solution,
1.0 % (w/v) was prepared in DW and used as an indicator. To prepare a
solvent for dissolving oil samples, 3 volumes of acetic acid were mixed
with 2 volumes of chloroform. The 0.01 M solution of sodium thiosulfate
was prepared by transferring the appropriate weighted mass of the
Na2S2O3 to a 100 ml volumetric flask and diluting it to the mark with
freshly boiled and cooled DW.
A total of 99 oil were used in this study. Among these, 80 samples
were used for deriving calibration curve and 19 samples were used for
method validation). Some samples were received from the partner lab­
oratory of Hormozgan Standard Department (4 samples of corn oil, 4
samples of sesame oil, 5 samples of sunflower oil, 5 samples of frying oil,
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Food Chemistry 403 (2023) 134345
and 9 samples of mixed oils (prepared from mixing of different oil
samples, e.g., mixing of canola with sunflower or soybean), 4 samples of
coconut oil, 4 samples of palm kernel oil and 2 samples of cocoa butter
substitute (CBS)). In addition, some samples were collected from local
supermarkets (9 samples of corn oil, 5 samples of olive oil, 5 samples of
sesame oil, 6 samples of frying oil, 14 samples of sunflower oil and 5
samples of canola oils). Moreover, 18 cold-press oil samples that are
directly extracted from the beans without any further processing were
obtained from Shiraz oil refineries in Shiraz city. The samples were from
different beans including Hazelnuts, Walnuts, Almonds, Olives, Sesame
and Sunflower oil).
2.2. Sample preparation
To measure the PV, the oil samples were made as uniform as possible
by shaking them in a closed container.For solid samples, the oil was first
melted by heating in the oven, controlled at a temperature at least 10 ◦ C
above the melting point of the particular oil. After heating, in case of
turbidity, the oil sample was filtered using a filter funnel (ISO, 2003).
2.3. Fabrication of the PAD and design principles
The PADs were prepared using a puncher in a circle-like pattern. The
available puncher had a diameter of 5 mm and so, the future designs
were based on this dimension. The principal of the detection method
utilized in this assay was the reaction of the lipid hydroperoxides with an
iodide solution, which forms molecular iodine. The amount of the
formed iodine was detected by its interaction with starch. The color
intensity of the iodine–starch complex on the paper substrate was then
correlated with the PV index pre-determined by a standard method (e.g.,
ISO, 2017). The basic principle of this was similar to the titrimetric
method of PV determination.
To do the assay on a paper microfluidic, two separate zones for the
reaction of the sample with potassium iodide and interaction of the
liberated iodine with starch are required. Since the reaction between
iodide and hydroperoxides is not instantaneous and needs time to
complete, the assay cannot be achieved on a two-dimensional paper
microfluidic, like a lateral flow assay. Actually, the movement of solu­
tions through hydrophilic channels in paper-based microfluidics usually
takes around one minute or lesser. Therefore, it was decided to use 3D
microfluidic. In its simplest form, a two-layer device was fabricated.
E. Ghohestani et al. Food Chemistry 403 (2023) 134345

Fig. 2. Effect of (A) starch concentration, (B) number of saturated-KI layer and (C) the sample volume, (D) reaction and detection times on the qualitative deter­
mination of peroxide value of the oil samples. Error bars refer to standard deviation (n = 5).

Fig. 3. Correlation between PVs obtained using the official method and obtained color intensity of colorimetric method. The brown band represents sensor’s
color changes.

Two sheets of filter paper containing potassium iodide and starch so­ and used. The metallic mold provides reproducible force when con­
lutions, separately, were cut in circular form and installed on top of each necting the paper layers and in the same time it protects the reaction
other. To maintain the layers attached and aligned, a metallic mold with from the interfering effect of ambient light.
cylindrical holes (see Supplementary Material Fig. S1) was fabricated To deposit reagents on the paper, two methods were examined (i)

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E. Ghohestani et al. Food Chemistry 403 (2023) 134345

Table 1 2.5. PV determination using an official standard method

Compartion of PV obtained using the ISO titrimetric method and the proposed
PAD method. The reults have been reported as mean of four replicated meas­ The PV of oil samples was determined according to ISO, 2017.
urments (meqO2/Kg oil) ± standard deviation. Approximately, the oil sample was weighted into the glass-stopper
Sample PAD Standard method Absolute error Erlenmeyer (5.0 g for peroxide value from ˃1 to 30 and 10.0 g for
Corn oil 3.53 ± 0.56 3.95 ± 0.31 0.42 peroxide value from 0 to 1) and dissolved in 50.0 ml solution of glacial
Coconut oil 1.16 ± 0.47 1.61 ± 0.14 0.45 acetic acid: chloroform with the ratio of 3:2. Then, 0.5 ml saturated-KI
Coconut oil 0.41 ± 0.08 0.40 ± 0.05 0.01 solution was added, and the mixture was shaken for 1 min. Immedi­
Palm oil 1.17 ± 0.31 1.00 ± 0.06 0.17 ately was added 100 ml of DW and after adding 0.5 ml starch solution,
Sunflower oil 4.48 ± 0.71 5.68 ± 0.43 0.20
Canola oil 5.92 ± 0.42 5.31 ± 0.41 0.61
the mixture was titrated with standard sodium thiosulfate 0.1 M. For
Frying oil 2.17 ± 0.63 2.67 ± 0.2 0.50 each sample, a separate blank analysis was also performed. PV was
Corn oil 3.57 ± 0.64 4.35 ± 0.21 0.78 determined using the following equation:
Frying oil 5.52 ± 0.32 5.60 ± 0.25 0.08
Olive oil 18.97 ± 2.14 22.94 ± 1.12 3.97 (V − V0)C*1000
PV = ( )
Canola oil 3.42 ± 0.36 3.56 ± 0.29 0.14 m
Canola oil 1.81 ± 0.28 1.62 ± 0.18 0.19
Canola oil 2.10 ± 0.19 2.23 ± 0.21 0.13 Where m was the mass of oil (g), C was the concentration of sodium
Olive oil 4.65 ± 0.31 3.90 ± 0.28 0.75 thiosulphate (M), V represented the volume of sodium thiosulfate used
Almond oil 25.16 ± 1.34 26.5 ± 0.94 1.34
Corn oil 15.63 ± 1.98 16.52 ± 1.05 0.89
by the sample and V0 was the volume (mL) of sodium thiosulfate used by
Sunflower oil 29.32 ± 1.45 31.40 ± 1.85 1.08 blank.
Sunflower oil 0.8 ± 0.07 0.55 ± 0.03 0.25
Mixed oil 11.12 ± 1.21 12.08 ± 0.95 0.98 3. Results and discussion

3.1. Optimization of the assay

firstly punching the paper and pouring small volumes of reagent on the
surface of the punched paper and (ii) firstly immersing a large piece of
To obtain reproducible and accurate results, the experimental con­
paper in the reagent’s solutions and then punching the paper after
ditions were optimized. The effects of 5 parameters including concen­
removing out from the solutions and drying. It was found that the second
trations of starch solution, the number of paper layers of KI solution, the
procedure provided a more uniform distribution of the reagents on the
volume of sample solution, the reaction time, and the measurement
paper surface and therefore more homogeneous color distribution was
time, which are supposed affecting on the device’s performance, were
observed when iodine interacted with starch.
considered. Each treatment was run 5 times. As a response variable, the
The reaction zone was prepared by immersing the Whatman paper in
average of color intensities for replicated measurements and the stan­
the freshly saturated KI solution for 30 s and then drying in dark at room
dard deviation of the measurements were considered. It should be noted
temperature (25 ◦ C). Then, the prepared circular patterns were stored in
that there is a reverse relationship between the color values and the level
a sample tube and wrapped with aluminum foil. In the same way, to
of peroxidases in oils. Higher peroxidases produce more iodine and
prepare the detection zone, the paper was immersed in the starch so­
therefore darker color spots are formed in the detection layer. Darker
lution, dried, and finally punched.
colors have lower values of RGB color elements. The white paper
The whole assay procedure is shown schematically in Fig. 1. The
without any color spot has color intensities of 255 for all RGB values. So,
fabricated mold is able to hold 5 devices simultaneously. The corre­
desirable experimental conditions possess simultaneously lesser color
sponding shafts on the complementary sheet of the mold push the device
values and lower standard deviation among 5 repeated measurements.
layers such that the solution inside them can communicate. When paper
To study the effect of starch concentration as the indicator, starch
layers were held into the mold holes, the sample solutions were added.
solutions in the concentration range of 0.5–5.0 % were used whereas
Afterward, DW was added and the complementary sheet of the mold was
other parameters were kept constant (one paper layer of saturated KI,
installed. After completion of the reaction, the mold was opened, the
40 μL of the sample solution, and 10 min for both reaction and detection
paper sheets were removed, and the image of the detection layer was
times). As shown in Fig. 2A, lesser amounts of starch resulted in more
recorded using a flatbed scanner.
reproducible data. However, better color intensities were observed for
starch concentration of 1 %. This concentration also represented
2.4. Colorimetric detection of PV using the PAD
acceptable reproducibility, which is better than larger concentrations of
starch. Therefore, a starch concentration of 1 % was used in the detec­
Initially, two layers of the pad were placed inside the mold (Fig. 1).
tion layer.
The weighted amount of the oil sample (0.25 or 0.50 g) was dissolved in
Since in the iodometric titration, KI must be existing in the excess
250.0 μL of the previously prepared solvent (a mixture of 3 volumes of
amount, it was attempted to use more than one paper layer of KI to be
acetic and 2 volumes of chloroform). A 35.0 μL aliquot of this solution
ensured that the amount of saturated KI for reaction with hydroperoxide
was dropped onto the device. The device’s cap was then closed, and the
was sufficient. Since saturated KI solution was used it was not possible to
liquid was allowed to transport to the detection zone through vertical
change the amount of KI by changing its concentration. Instead, the
flow in a sufficient time (10 min). In the reaction zone, the lipid hy­
number of paper layers pre-deposited by saturated KI solution was
droperoxides is reacted with the deposited iodide ions on the paper to
changed. As shown in Fig. 2B, the sensor’s response was not affected
form the iodine molecules. Afterward, by the addition of 10 μL of DW,
significantly by the number of KI layers suggesting using just one paper
the produced iodine in the reaction zone was moved to the detection, on
layer of saturated KI solution was enough.
which starch was pre-deposited. Changes in the color of the detection
The third optimized parameter was the sample volume. The required
zone were captured by a flatbed desktop scanner and the images were
volume of the sample solution should be large enough to pass the re­
digitized into the red, green, and blue (RGB) color intensity using
agent layer and then wet the detection layer completely. As shown in
ImageJ software. The Euclidean norm of the color elements
√̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅ Fig. 2C, the sensor response was enhanced by increasing the sample
( R2 + G2 + B2 ) was used as an analytical signal to provide the cali­
volume from 10.0 to 30.0 μL. Afterward, no significant changes were
bration curve and estimate PV.
observed.
As explained, the detection principle in this essay is composed of two

4
E. Ghohestani et al.
Fig. 4. The PCA score plots for classification of different ranges of peroxide value (A) according to the Iranian national standards and (B) Codex standard.
consecutive steps (i) reaction of lipid hydroperoxides with iodide solu­
tion and production of iodine, called reaction step, and (ii) detection of
the liberated iodine using starch, called detection step. The required
time for the completion of reactions in both steps needs to be optimized.
Since these reaction steps are assumed to be dependent, an interaction
between the parameters of reaction times was assumed. Therefore, these
parameters were optimized using a 3-level full factorial design, each
with levels of 5, 10, and 15 min. The results are given in Fig. 2D. As seen,
better responses were obtained at longer times (15 min for both re­
actions). However, because of water evaporation, this reaction condition
was associated with lower reproducibility. On the other hand, the re­
action times of 10 min (for both steps) represented an appropriate
response and at the same time much better reproducibility.
To be ensure about completeness of the reaction, the detection time
was considered fixed at 10 min, and variable reaction times of 5, 10, 15,
and 20 min were used. As shown in Fig. S2, no significant difference in
the color intensity of the PAD was observed in reaction times longer than
10 min.
In summary, the selected optimized condition is as follows:
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Food Chemistry 403 (2023) 134345
(i) The reaction and detection layers are soaked in saturated KI and
1 % starch solutions, respectively.
(ii) For both reaction steps, one paper layer is used.
(iii) A sample volume of 30.0 μL is used and after 10 min, 10.0 μL of
water is added.
(iv) The color of the detection zone is imaged 10 min after water
addition.
3.2. Calibration and validation
Under the optimum condition, 80 different oil samples belonging to
11 groups of oils (olive, corn, sesame, sunflower, frying, mixed, coconut,
palm kernel, cocoa butter substitute (cbs), and cold press oils (hazelnuts,
walnuts, almonds, olives, sesame and sunflower) were used for training
the sensor with 4 replicate for each sampels. The degree of oxidation of
oil samples (PV) was analyzed using the standard method. Then, the
measured PVs were correlated with the respective color intensity of the
colorimetric sensor. As shown in Fig. 3, there is an inverse logarithmic
relationship between the PV and the color intensity. Obviously, the
observed logarithmic relationship is valid in the wide range of PV
E. Ghohestani et al.
Fig. 5. Photograph of the paper devices for investigating the stability of PAD. The images in every frame represent repeated measurements.
(0.01–30.00 meq/Kg) and interestingly the relationship is independent
of the type of oil. The inverse logarithmic relationship implies that the
sensor possesses higher sensitivity at lower PVs, which is a fortune event
since the sensor is able to discriminate oils of very small PV.
Limit of detection (LOD) was calculated according to the IUPAC
recommendation by replicate analysis of solvent without added oil as
y +3s
blank solution (LOD = b m b ), where yb andsb are the average and stan­
dard deviation of blank measurements, respectively. Since the calibra­
tion curve is logarithmic, the first few data points were fitted to a linear
model and initial slope (m) used. In this way, LOD was calculated as
0.015 meq/Kg. The precision of the method, as relative standard devi­
ation (RSD), was obtained by analyzing oil samples of different peroxide
value. RSD values in the range 0.3–10.0 % were obtained.
At the bottom of Fig. 3 are shown the scanned images of the sensor
for different PVs in the range of 0.01–30 meq/Kg. These images can be
used a color bar for semi-quantitative estimation of PV by naked eye
without needing a scanner or image analysis.
To validate the sensor accuracy, 19 new oil samples were analyzed
by the sensor with 4 replicate measurements and their PV values were
estimated using the calibration model in Fig. S3. Then, their estimated
PV by the sensor was compared with their PV measured by ISO titri­
metric method. The estimated and measured values are listed in Table 1
and are correlated in Fig. S2. There is observed a close agreement be­
tween the PV indices measured by the standard method and estimated
by our sensor. The correlation between two values follows a linear
regression equation of intercept and slope of 0.09 and 1.09, respectively,
which are close to the ideal values of zero and one, respectively.
In the next step, instead of converting the R, G, and B values of the
sensors’ images to a single digit (by calculating their Euclidean norm),
they were used to represent every sensor in a 3-dimensional vector
space. Then, a data matrix was provided by stacking the color element
vectors of the sensors of all samples sizing (80 × 3). This data matrix was
then subjected to multivariate data analysis.
According to the maximum level of peroxide value approved for the
cold press oils and edible oils in the Iranian national standards, the
maximum level of PV can be classified into four ranges, 0 < PV ≤ 1, 1 <
PV ≤ 2, 2 < PV ≤ 5 and 5 < PV ≤ 10 meq/Kg for the edible oils at the
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Food Chemistry 403 (2023) 134345
time of production, free at the port of shipment, consumable limit,
respectively (INSO, 2014). Also, according to the Codex standards, the
maximum level of PV for refined and cold press oils is PV ≤ 10 and PV ≤
15 meq/Kg respectively. (Alimentarius, 2019) In this regard, multivar­
iate data analysis was employed to device classification models that are
able to cluster oil samples into groups based on the above defined
standards. The provide data matrix of color intensity values was first
subjected to principal component analysis (PCA) as an unsupervised
exploratory data analysis. Then, the extracted principal components
(PCs) were fed into linear discriminant analysis to build classification
models based on the Iranian national standard (4-class classification
model) and Codex standard (2-class classification model). The results
are given in Fig. 4. As observed, qualitative discrimination between
different ranges of peroxide value of oils has been achieved. However,
some overlap is observed between classes of the Codex standard. The
overlapped data correspond to the PVs of 10.0 ± 1. The classification
accuracies for training and cross-validation were 90 % and 93 % for the
Iranian national standard and 89 % and 91 % for the Codex standard,
respectively. The results of these analyses suggest the design PAD can be
used as a simple device for fast screening of vegetable oils.
3.3. Stability
Storage stability is an essential parameter for point-of-use and rapid
detection of PV in the on-site analysis. Since light and oxygen cause the
oxidation of iodide ions, sensors’ stability was investigated in the dark
condition. The fabricated devices were immediately stored in a sample
tube, which was wrapped with aluminum foil, and were kept at room
temperature (25 ◦C) for 3 months. Six different oil samples were then
analyzed by the freshly prepared PADs and the 30-day old PADs. As
shown in Fig. 5, there are no significant differences between the images
of the fresh and aged sensors. The color intensities of the PADs indices
measured by both types of PADS are given in Table S1. Analysis of
variance (ANOVA) and t-test were used to evaluate the statistical dif­
ferences between the fresh and aged PADs. Both methods supported no
significant differences between the results of the aged and fresh PADs at
95 % confidence level, suggesting the stability of the sensor for 30 days.
E. Ghohestani et al.
Moreover, as given in Table S1, the reproducibility in color intensity of
the repeated measurements remains the same for the aged PADs relative
to the fresh one (for all oil samples, the relative standard deviations are
always less than 4 %.
3.4. Effect of blank experiment
As noted previously, the reaction of iodide with hydroperoxide may
be interfered by oxygen and light. To investigate the effect of these
parameters on the performance of our device, a blank experiment, where
30.0 µL of distilled water was added instead of oil, was run. Firstly, the
blank’s response was the same for all oil samples and no color change
caused by the oxidation of iodide was observed. Secondly, as shown in
Fig. S4, the original images and those calculated after subtracting the
blank image look very similar. Indeed, the calibration curves plotted
based on the original images and the blank-subtracted images repre­
sented no significant differences in their statistical parameters (Fig. S5).
These observations suggest that the response of the device is indepen­
dent of blank and the images of the sensor can be directly used for
estimation of PV.
4. Conclusion
Overall, we have designed a paper-based device for the determina­
tion of PV in the refined and cold press oil samples based on the iodo­
metric reaction. Under optimal conditions, the device response
represented a clear and well-defined inverse logarithmic relationship
with the PV indices measured by the conventional iodometric titration
(ISO-3960) with a working range of 0.01–30.0 meq/Kg. Also, semi-
quantitation of PV was achieved by multivariate analysis of the color
values. The peroxide measurements by our method offer a faster
analytical procedure that does not need glassware and skilled personnel
and consumes very low amounts of reagents and solvents. The PADs
were stable for months when kept under dark conditions and at room
temperature (25 ◦ C). The time needs for analysis by PAD (20 min) is
slightly lower than that of the conventional. However, the PAD method
offers some other advantages like (i) it needs much less reagents and
solvents, (ii) the device is pre-calibrated and hence it can be used for on-
site analysis without need handling reagents/solutions.
CRediT authorship contribution statement
Elham Ghohestani: Investigation, Methodology, Writing – original
draft. Javad Tashkhourian: Methodology, Supervision. Bahram
Hemmateenejad: Conceptualization, Supervision, Writing – review &
editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgments
The authors thank the Shiraz University Research Council for sup­
port of this work.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
Food Chemistry 403 (2023) 134345
org/10.1016/j.foodchem.2022.134345.
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