Food Chemistry 249 (2018) 154–161

Contents lists available at ScienceDirect

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

Analytical Methods

A biodegradable colorimetric film for rapid low-cost field determination of T

formaldehyde contamination by digital image colorimetry
⁎
Worawit Wongniramaikula,b,c, , Wadcharawadee Limsakula, Aree Chooduma,b
a
Integrated Science and Technology (Applied Chemistry/Environmental Management and Software Engineering) Research Center, Faculty of Technology and Environment,
Prince of Songkla University, Phuket Campus, Kathu, Phuket 83120, Thailand
b
Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand
c
Center of Excellence on Hazardous Substance Management (HSM), Bangkok 10330, Thailand

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

Keywords: A biodegradable colorimetric film was fabricated on the lid of portable tube for in-tube formaldehyde detection.
Biodegradable film Based on the entrapment of colorimetric reagents within a thin film of tapioca starch, the yellow reaction
Colorimetric sensor product was observed with formaldehyde. Intensity of the blue channel from the digital image of yellow product
Formaldehyde showed a linear relationship in the range of 0–25 mg L−1 with low detection limit of 0.7 ± 0.1 mg L−1. Inter-
Digital image colorimetry
day precision of 0.61–3.10%RSD were obtained with less than 4.2% relative error from control samples. The
developed method was applied for various food samples in Phuket and formaldehyde concentration range was
non-detectable to 1.413 mg kg−1. The quantified concentrations of formaldehyde in fish and squid samples
provided relative errors of −7.7% and +10.8% compared to spectrophotometry. This low cost sensor
(∼0.04 USD/test) with digital image colorimetry was thus an effective alternative for formaldehyde detection in
food sample.

1. Introduction spectrometry (GC–MS), and high performance liquid chromatography

Formaldehyde (HCHO) is a toxic substance, which can cause severe
health issues (Bunkoed, Davis, Kanatharana, Thavarungkul, & Higson,
2010; International Agency for Research on Cancer, 2004; WHO
Regional Office for Europe, 2001). It can irritate the eyes and nose,
damage the central nervous system, and cause immune system dis-
orders. Moreover, it has been classified as carcinogenic to humans by
the International Agency for Research on Cancer (IARC) (International
Agency for Research on Cancer, 2004). However, it is still widely used
as a precursor in many industries (Halvarsson, Edlund, & Norgren,
2008; Kim, 2009), e.g. textile, plastics, and wood industries, due to its
high reactivity and relatively low cost. It is also commonly used as a
preservative in medical laboratories, mortuaries, and consumer pro-
ducts. Formaldehyde has been used also inappropriately to extend the
shelf-life of food (Atahar, 2013; Riaz, Moin, Tasbira, Naz, & Kumar,
2011), as it was found in various foods at higher concentrations than
deemed natural (e.g. 1.8 ppm in squid and 1.0–2.4 ppm in prawn) (Clary
& Sullivan, 2001; World Health Organization, 2016). The liquid form of
formaldehyde, namely formalin (35–40% aqueous solution stabilized in
methanol), was commonly used for this purpose.
A wide range of analytical methods, e.g. gas chromatography-mass
(HPLC), have been reported for the determination of formaldehyde, or
formalin, in food (Wahed, Razzaq, Dharmapuri, & Corrales, 2016; Wang
et al., 2012; Yeh, Lin, Chen, & Wen, 2013; Zhu, Peng, Wang, Wang, &
Rui, 2012). However, these are analytical laboratory methods with
expensive and sensitive devices, not with robust portable field instru-
ments. A colorimetric sensor would be an advantageous choice for rapid
cost-effective on-site determinations of formaldehyde. This approach
would provide a number of advantages such as field portability, visual
qualitative feedback, rapid detection, and ease of handling (Maruo,
Nakamura, Uchiyama, Higuchi, & Izumi, 2008b; Wang et al., 2014). It
is typically based on entrapment or impregnation of a colorimetric re-
agent within various materials, such as a sol–gel matrix (Bunkoed et al.,
2010), porous glass (Maruo et al., 2008b), or molecularly imprinted
polymer (Feng, Liu, Zhou, & Hu, 2005). However, these types of sub-
strate tend to be non-degradable and of themselves contribute to en-
vironmental problems. The use of a biodegradable natural polymer as
the supporting material could eliminate this drawback and would be a
further asset in a simple and rapid colorimetric sensor. In this work, a
natural polymer that is inherently biodegradable (Chiellini & Solaro,
1996) has been used to fabricate the colorimetric sensor for for-
maldehyde detection. Starch was selected due to its natural availability,

⁎
Corresponding author.
E-mail address: worawit.won@phuket.psu.ac.th (W. Wongniramaikul).

https://doi.org/10.1016/j.foodchem.2018.01.021
Received 30 December 2016; Received in revised form 11 November 2017; Accepted 2 January 2018
Available online 03 January 2018
0308-8146/ © 2018 Elsevier Ltd. All rights reserved.
W. Wongniramaikul et al. Food Chemistry 249 (2018) 154–161

complete biodegradability (Araújo, Cunha, & Mota, 2004), low cost and
renewability (Zhang & Sun, 2004).
A colorimetric sensor is commonly used in conjunction with a
spectrophotometric method for the quantitative analysis of for-
maldehyde (Bunkoed et al., 2010; Fagnani, Melios, Pezza, & Pezza,
2003; Maruo et al., 2008b). This requirement of a spectrophotometer
could limit the field applications of an analytical method. However, in
recent years digital image colorimetry (DIC) has gained attention due to
its low cost in rapid real-time quantitative determinations (Choodum,
Boonsamran, NicDaeid, & Wongniramaikul, 2015a; Choodum,
Kanatharana, Wongniramaikul, & Nic Daeid, 2013; Choodum,
Kanatharana, Wongniramaikul, & NicDaeid, 2012; Choodum & Nic
Daeid, 2011; Choodum et al., 2014). The DIC is based on the analysis of
RGB data (Red Green Blue basic color) from common digital images of
the colorimetric product generated by a digital camera, such as a digital
single-lens reflex camera (Choodum & Nic Daeid, 2011; Choodum et al.,
2012), the built-in digital camera in a mobile phone (Choodum et al.,
2013), or a web camera (Choodum et al., 2015a)). During imaging
process, the reflected light from the object of colorimetric product is
passed through RGB filters and separated into three spectral ranges.
They were then detected and recorded as individual RGB values by the
image sensor located behind the filter. The RGB data allows re-
generating a subjectively faithful color digital image of the object.
When the digital color image is analyzed by a color analysis program,
e.g. Matlab (Lopez-Molinero, Liñan, Sipiera, & Falcon, 2010), Kylix
(Gaiao et al., 2006), Visual basic (Maleki, Safavi, & Sedaghatpour,
2004), or Adobe Photoshop (Choodum & Nic Daeid, 2011; Choodum
et al., 2012, 2013), the individual RGB image layers can be inspected.
These RGB data can be used as analytical data to produce a calibration
graph for quantitative analysis of the interested analyte. In this work,
DIC was used for the rapid quantification of formaldehyde in foods, in
conjunction with a biodegradable and portable sensor. Thus, the rapid
and simple quantitative analysis of formaldehyde was achieved on-site,
using an environmentally friendly colorimetric test kit and the digital
camera of a mobile phone.
2. Materials and methods
2.1. Materials
A stock solution of 5000 mg L−1 formaldehyde was prepared from
formaldehyde solution (37%w/v) (Supelco, Bellefonte, PA) in ultrapure
water (Barnstead EasyPure II, Thermo fisher scientific, OH). Standard
working solutions were prepared daily by diluting the stock solution to
appropriate concentrations in ultrapure water. Acetylacetone and am-
monium acetate were obtained from Sigma-Aldrich (St. Louis, USA),
while acetic acid solution was purchased from BDH Laboratory Supplies
(Poole, UK).
2.2. Preparation of the biodegradable colorimetric film
The biodegradable colorimetric film was prepared by mixing a
starch solution with a formaldehyde specific reagent. Tapioca starch
(0.5 g, from supermarket in Kathu, Phuket) was dispersed in ultrapure
water (10 mL). It was then heated on a hot plate (∼100 °C) under
continuous stirring until a clear viscous solution was obtained. After the
resultant solution was cooled down to room temperature, the colori-
metric reagent was added. Acetylacetone (0.75 mL), ammonium acetate
(1 g), and glacial acetic acid (0.1 mL) were added into 4 mL of the
starch solution and stirred for 6–8 min to obtain a homogeneous solu-
tion. The mixture (100 µL) was then transferred into the flat cap of a
centrifuge tube (1.5 mL) and heated at 120 °C for 30 min. This step
produced the thin film on the lid of the tube (Fig. 1a). After cooled to
room temperature, the lid was quickly closed onto the tube to avoid the
contamination, and then stored in a desiccator until further use.

Fig. 1. A biodegradable colorimetric film on the lid of a centrifuge tube: (a) schematic, (b) the film, and (c) colorimetric reaction products from various formaldehyde concentrations
(0–2500 mg L−1).

155
W. Wongniramaikul et al.
2.3. Characterization of the biodegradable colorimetric film
The morphology of the biodegradable colorimetric film was in-
vestigated using a scanning electron microscope (SEM) (Quanta400,
FEI, Czech Republic). Spectral analysis was performed by Fourier
transform infrared spectroscopy (FTIR) (Equinox55, Bruker, Germany)
over the wavenumber range from 4000 to 400 cm−1 at resolution of
4 cm−1, using KBr pellet samples.
2.4. Colorimetric test of formaldehyde using the biodegradable film
Standard working solutions of formaldehyde (0–2500 mg L−1) were
prepared by diluting a stock solution (5000 mg L−1) with ultrapure
water. One milliliter of each standard solution was transferred into a
centrifuge tube with the biodegradable film on the lid and mixed by
shaking for 1 min. The resultant yellow products in the centrifuge tubes
were then photographed for six times using the built-in digital camera
of an iPhone 6.0. Three colorimetric films were exposed to each cali-
bration concentration of formaldehyde. The average intensities of the
red, green and blue colors (RGB values) across six images from three
sensors at each formaldehyde concentration were investigated using an
in-house RGB analysis program.
2.5. RGB imaging system and procedure
A custom-built photographic box (Choodum, Kanatharana,
Wongniramaikul, & NicDaeid, 2016) was used throughout the experi-
ment to eliminate any interference from ambient environmental light.
Three sensors were hung at the top of the box. The front wall of the box
had a slot for the mobile phone (iPhone 6.0) with an opening for the
camera lens, so the colorimetric product could be photographed under
reproducible illumination.
The digital camera of the iPhone 6.0 was set to flash off and auto-
matic white balance, with the high dynamic range (HDR) off. Each
image was 1.18 MB (3264 × 2448-pixel) size when saved in JPEG (24-
bits) format to the iPhone’s memory. The images were transferred to a
computer and their RGB data were analyzed with an in-house RGB
analysis program (Choodum et al., 2016). In brief, an initial
50 × 50 pixel image patch from one of the six recorded images of three
sensor films in each test was manually selected for color analysis. An
equivalent image patch of the remaining five digital images was then
automatically selected by the image analysis program. The average RGB
values from all six images for each of the three sensors (18 values in
total) for each standard concentration were used as a single data point
to establish calibration curves.
2.6. Analytical performance and method validation
Analytical performance, i.e. sensitivity, linearity, linear range, limit
of detection (LOD), and limit of quantification (LOQ) were investigated.
LOD was calculated using an IUPAC method (Taverniers, De Loose, &
Van Bockstaele, 2004), i.e. LOD = 3.3SBl/S where SBl is the standard
deviation of blank signal from 10 analyses and S is the slope of the
calibration curve. LOQ was calculated using the same method, but
being equal to the blank signal plus ten standard deviations of the
blank. Precision is expressed as percentage relative standard deviation
(%RSD) for each color component, over six images each of three sensor
films (n = 18). The accuracy was evaluated by analyzing the known
concentration of formaldehyde standard (7.5 mg L−1) against the es-
tablished standard curve.
2.7. Quantification of formaldehyde in foods in Phuket
Fresh foods including fish, shrimp, squid, tomato, and mushroom
were collected from two fresh markets in Kathu and Samkong, Phuket,
Thailand on October 2016. The same types of foods together with
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Food Chemistry 249 (2018) 154–161
frozen octopus were also collected from two supermarkets within
nearby area in the same period. All collected samples were tested using
the biodegradable colorimetric films coupled with the DIC and com-
mercial formalin test kits (commonly used by Thai Public Health offi-
cers) within an hour after collection. Fish and squid samples were also
analyzed using spectrophotometer for comparison.
The biodegradable colorimetric films were freshly prepared using
optimum conditions for real sample analysis in the same day of sample
collection and analysis. Nine sensors were randomly selected and tested
with formaldehyde solution (10 mg L−1) using the optimum conditions
of DIC before real sample analysis to re-check their accuracy and pre-
cision which provided < 2.5%RSD and < 4.2% relative errors.
The collected food samples (100–400 g) were immersed in ultrapure
water (50 mL/200 g) for approximately 15 min in zip locked plastic bag.
The water samples were then tested with the film and the formaldehyde
concentrations were quantified using DIC. The foods were then washed
by water until clean and obtain ready-to-cook samples before immersed
in water for a further 15 min. Water samples were then tested by the
same procedure again to obtain the formaldehyde concentrations of
ready-to-cook samples.
3. Results and discussion
3.1. Preparation of the biodegradable colorimetric film and colorimetric test
with formaldehyde
The biodegradable colorimetric film was prepared by the entrap-
ment of colorimetric reagent within a thin film of natural polymer. The
reagents, which can selectively react with formaldehyde, were mixed
with the starch solution before casting. Various types of starch, i.e. ta-
pioca starch, corn starch, rice flour, and glutinous flour, were in-
vestigated as the natural polymer substrates that entrap the colori-
metric reagents. Tapioca starch provided a clear homogeneous film
with good temperature tolerance during casting, while other types
provided brown film, so it was selected for further use as a supporter.
Optimization of the film fabrication was done in order to obtain a
biodegradable colorimetric film with good characteristics and the best
performance for formaldehyde detection. The use of 5% (w/v) tapioca
starch solution (investigated over the range from 2.5 to 10%) for 4 mL
water (tested 1–10 mL) with 0.75 mL acetylacetone (tested
0.25–1.0 mL), 0.1 mL glacial acetic acid (tested 0.1–0.6 mL), and 1 g
ammonium acetate (tested 0.5 to 1.25 g) provided clear homogeneous
thin films (Fig. 1b) that produced the darkest yellow product of for-
maldehyde. The cost of each film at these conditions was only 0.30 THB
(∼0.008 USD), while the cost of overall sensor (film and centrifuge
tube) was 1.30 THB (∼0.04 USD) which are cheaper than the costs of
alternative detection methods. These conditions were thus used as the
optimum conditions to synthesize the low-cost biodegradable colori-
metric film for formaldehyde detection. The good precision of
0.75–1.52% RSD was obtained with three synthesis batches using these
conditions. The film also provided good stability (with 2.04%, 2.92%
and 0.96% change in red, green and blue intensity, respectively) after
storage in a refrigerator for three months.
Colorimetric test of formaldehyde using the biodegradable colori-
metric film was based on the Hantzsh reaction (Bunkoed et al., 2010;
Maruo, Nakamura, & Uchiyama, 2008a; Nash, 1953) between the en-
trapped colorimetric reagents within the film and formaldehyde. The
entrapped chemical reagents (acetylacetone, ammonium acetate, and
acetic acid) were dissolved into the sample solution containing for-
maldehyde, allowing the reactions. A yellow product was produced by
the reaction of formaldehyde and β-diketone under mild conditions.
Acetylacetone was selected as the β-diketone, because it produces more
intense color and reacts faster than methyl acetoacetate (Bunkoed et al.,
2010). Darker yellow products were obtained on increasing the for-
maldehyde concentrations (Fig. 1c).
W. Wongniramaikul et al.
Fig. 2. SEM image of the biodegradable colorimetric film at (a) 1500X, and (b) 10,000X, and of the starch film without reagents at (c) 10,000X, and (d) 25,000X.
3.2. Characterization of biodegradable colorimetric film
The SEM images of the biodegradable colorimetric film revealed a
homogeneous distribution of colorimetric reagents within the film
(Fig. 2a and b). In contrast, the film without colorimetric reagents (a
blank) had a smooth surface (Fig. 2c and d). As there were actually 3
colorimetric reagents, i.e. acetylacetone, acetic acid, and ammonium
acetate, different sizes of entrapped particles were observed.
FTIR spectrum of the biodegradable colorimetric film is presented in
Fig. 3. The large bands observed at 3214 and 2927 cm−1 were char-
acteristic peaks of the tapioca starch (Ali et al., 2013; Sacithraa,
MadhanMohan, & Vijayachitra, 2013), which are assigned to OeH and
CH2 symmetrical stretching vibrations, respectively. However, the OeH
band for the biodegradable film was shifted when compared to the pure
tapioca film that presented the band at 3395–3423 cm−1 (Ali et al.,
2013; Sacithraa et al., 2013; Sutjarittangtham et al., 2014). This might
be caused by the OeH stretching of acetic acid commonly presented at
3200–3000 cm−1 and/or NeH vibrations of ammonium acetate
(3000–2500 cm−1). The absorption bands from 1149 to 934 cm−1 are
assigned to CeO vibrations of amylopectin from tapioca (Deeyai,
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Food Chemistry 249 (2018) 154–161
Suphantharika, Wongsagonsup, & Dangtip, 2013; Sutjarittangtham
et al., 2014), and the peaks at 857 and 759 cm−1 are attributed to
skeletal stretching of starch. The absorption peaks observed at 1700 and
1407 cm−1 are assigned to the vibrations of acetic acid, while the peaks
at 1553 and 1263 cm−1 are attributed to the vibrations of acetylacetone
in enol form (Lozada-Garcia, Ceponkus, Chin, & Chevalier, 2011; Sohn
& Lee, 1997)
3.3. Digital image colorimetry for formaldehyde quantification
The colorimetric products from the reaction between formaldehyde
and the entrapped reagents within the biodegradable film were pho-
tographed using the built-in digital camera of an iPhone 6.0. All digital
images were systematically analyzed by a custom-built RGB analysis
program to obtain RGB values as analytical data. The relationship be-
tween formaldehyde concentration and RGB values revealed higher
intensities in the green and the red channel than in the blue channel
(Fig. 4a). This was expected from the subjective color of the colori-
metric product, i.e. a yellow product would reflect more red and green
light (thus appear yellow) than blue.
W. Wongniramaikul et al.
The intensities of red and green channels tended to be constant with
increasing formaldehyde concentration, while the intensity of the blue
channel was decreased. These results indicated that the yellow products
absorbed light in the blue range. When the more colorimetric products
were produced at higher analyte concentrations, the darker yellow
product absorbed more blue light reducing its intensity. This was in
good agreement with the calculated molecular absorption (Choodum
et al., 2012, 2013, 2014) based on formaldehyde concentration. The
blue channel (400–500 nm) had the strongest absorbance (Fig. 4b) and
Fig. 4. Relationships between formaldehyde concentrations and (a) individual RGB values, (b) calculated absorbance, (c) individual RGB values with blank subtraction, and (d) calculated
absorbance with blank subtraction.
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Food Chemistry 249 (2018) 154–161
Fig. 3. FTIR spectra of the biodegradable colorimetric film.
was increased with formaldehyde concentration. This was also in good
agreement with spectrophotometric results, which provided the max-
imum absorption at 462 nm, although prior studies have reported also
410 nm (Bunkoed et al., 2010) and 407 nm (Maruo et al., 2008a).
The relationships between the blank subtracted RGB values and the
formaldehyde concentration were also assessed on intensity (Fig. 4c)
and calculated absorbance (Fig. 4d) scales. Blank subtracted intensity
and absorbance of the blue channel increased with formaldehyde con-
centration, while these are nearly constant for the red and green
W. Wongniramaikul et al. Food Chemistry 249 (2018) 154–161

Table 1
Calibration equation and analytical performance of the biodegradable colorimetric film and DIC in formaldehyde quantification.

Relationship Calibration equation Linearity Linear range LOD LOQ

(y = a.u., x = mg L−1) (R2) (mg L−1) (mg L−1) (mg L−1)

IB y = −(3.0 ± 0.1)x + (161 ± 2) 0.9946 0–25 0.7 ± 0.1 2.3 ± 0.1

AB y = (0.0107 ± 0.0003)x + (0.194 ± 0.004) 0.9972 0–25 0.46 ± 0.06 1.41 ± 0.06
|IB − IBBlank| y = (2.86 ± 0.07)x + (5 ± 1) 0.9987 1–25 0.78 ± 0.04 2.38 ± 0.04
|AB − ABBlank| y = (0.0106 ± 0.0004)x + (0.003 ± 0.006) 0.9967 1–25 0.47 ± 0.09 1.42 ± 0.09

channels. These indicated that the greatest difference between the color
of the blank and products with increasing formaldehyde concentrations
caused by the blue light. The relationships of the blank subtracted blue
intensity (|IB − IBBlank|) and/or blue absorbance (|AB − ABBlank|) may
be beneficial with colored samples that interfere with observing the
colored product (Choodum, Kanatharana, Wongniramaikul, &
NicDaeid, 2015b; Choodum et al., 2016), e.g. shred ginger with pale
yellow appearance.
3.4. Quantitative analysis of formaldehyde using the biodegradable
colorimetric film and digital image colorimetry
The quantitative analysis of formaldehyde using the biodegradable
colorimetric film and digital image colorimetry was achieved by using a
linear portion of the relationships described in Section 3.3. As shown in
Table 1, both the intensity and the calculated absorbance the blue
channel had linear responses in the range from 0 to 25 mg L−1 of the
analyte with good linearity (R2 > 0.9946), while the blank subtracted
measurements provided better linearity (R2 > 0.9967) in the con-
centration range of 1–25 mg L−1. These could all be used as standard
curve for quantifying formaldehyde. Because the blue intensity was
obtained directly from the analysis of a digital image, while the mole-
cular absorption was calculated from the intensity, the linear relation-
ships of the blue intensity with (|IB − IBBlank|: yB = (2.86 ± 0.07)
x + (5 ± 1)) and without blank subtraction (IB: yB = −(3.0 ± 0.1)
x + (161 ± 2) were thus recommended for quantification of for-
maldehyde.
3.5. Analytical performance and method validation
The analytical performance of the biodegradable colorimetric film
and digital image colorimetry for formaldehyde quantification was also
shown in Table 1. LOD were in the range of
0.46 ± 0.06–0.78 ± 0.04 mg L−1, while the LOQ were in the range of
1.41 ± 0.06–2.38 ± 0.04 mg L−1. These performance characteristics
were effective to determine the formaldehyde contaminant in foods, as
the World Health Organization has reported the following natural
contents of formaldehyde: fruits and vegetables from 3.3 to 60 mg kg−1,
meat from 8 to 20 mg kg−1, milk from 1 to 3.3 mg kg−1, and fish from 1
to 98 mg kg−1 (WHO Regional Office for Europe, 2001; World Health
Organization, 1989; Yeh et al., 2013). Good intra-day and inter-day
precisions (n = 3) were obtained in the ranges 0.33–2.58% and
0.61–3.10%RSD. When control samples were tested with the proposed
method, the relative errors of +4.1% and +4.2% were obtained by
using the IB and |IB − IBBlank| equations, respectively, indicating a good
accuracy. In addition, the proposed method was in good agreement
with the spectrophotometric method (the relative error of −7.7% for
fish and +10.8% for squid samples).
3.6. Quantification of formaldehyde in foods in Phuket
Three vegetables (tomato, shredded ginger and jew’s ears mush-
room), three fresh seafood samples (shrimp, fish, and squid), and one
frozen octopus were collected from two supermarkets and two fresh
markets in Phuket. The freshly prepared biodegradable colorimetric
film was applied to quantify the formaldehyde concentration in foods
by coupling with DIC and using the standard curve for |IB − IBBlank|
(yB = (2.86 ± 0.07)x + (5 ± 1)). The results were shown in Table 2.
Concentration of formaldehyde in all the collected samples was in the
range considered natural, or below the limits reported as potentially
natural. For example, the formaldehyde range from 0.1 to 1.1 mg kg−1
found in tomato compares well to the natural occurrence from 5.7 to
13.3 mg kg−1 (World Health Organization, 2016), while the range from
0.1 to 0.4 mg kg−1 found in fish is below the potential natural occur-
rence reported by WHO in 1989 (1–98 mg kg−1, World Health
Organization, 1989) and Jaman et al. in 2015 (1.45–2.60 mg kg−1,

Table 2
Quantification of formaldehyde in foods on Phuket, using the biodegradable colorimetric film and DIC.

Type of food Water sample Concentration of formaldehyde (mg kg−1) in food collected from

Fresh market A Fresh market B Supermarket A Supermarket B

Tomato Before cleaning 0.488 0.668 1.413 0.594

Ready to cook 0.491 0.136 0.987 1.087

Mushroom Before cleaning 0.565 0.689 0.667 0.552

Ready to cook 0.534 nd* 1.412 0.976

Shred ginger Before cleaning 0.135 0.352 – –

Ready to cook 0.326 0.044 – –

Fish Before cleaning 0.368 0.334 0.369 0.121

Ready to cook nd 0.366 0.277 0.260

Shrimp Before cleaning 0.599 nd 0.143 0.081

Ready to cook 0.524 nd 0.691 0.132

Squid Before cleaning nd nd nd 0.962

Ready to cook nd nd nd 1.195

Frozen octopus Before cleaning – – 0.326 0.972

Ready to cook – – 0.656 1.189

* nd: not detectable.

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W. Wongniramaikul et al.
Jaman, Hoque, Chakraborty, Hoq, & Seal, 2015). It seems that fresh
vegetable samples, both tomato and mushroom, from the supermarket
contained more formaldehyde than those from the fresh market. The
obtained results were agreed with those from the commercial test kits
commonly used by Thai Public Health officers, which showed negative
results to all collected samples. Concentrations of formaldehyde in all
samples quantified by the proposed method were in the range of non-
detectable to 1.413 mg kg−1 which were lower than detection limit of
commercial test kits (5 mg kg−1). The proposed method was also in
good agreement with the spectrophotometric method as the relative
error of −7.7% for fish and +10.8% for squid samples were obtained.
4. Conclusions
Rapid determination of formaldehyde in foods was achieved by
using a novel biodegradable colorimetric film in conjunction with di-
gital image colorimetry. This made the detection of formaldehyde
successful by using an environmentally friendly colorimetric sensor.
The biodegradable film was fabricated onto the lids of small centrifuge
tubes, to which the sample solutions could be directly added for in-tube
detection and easily portable. The cost of each film was only 0.30 THB
(∼0.008 USD), while the cost of the sensor (film and centrifuge tube)
was 1.30 THB (∼0.04 USD), both of which are below per sample costs
of alternative approaches. When the developed biodegradable colori-
metric film was used in combination with digital image colorimetry
(DIC), it provided an ideal novel platform for the rapid quantitative
analysis of formaldehyde. The DIC approach requires only a digital
camera, here one in a daily used mobile phone, instead of a spectro-
photometer that is commonly used in quantitative colorimetry. The
developed method provided linear relationships in the range of
0–25 mg L−1 (R2 > 0.995) with low detection limit of
0.46 ± 0.06–0.78 ± 0.04 mg L−1. Method validation also showed <
3.10%RSD inter-day precision and < 4.2% relative error in for-
maldehyde determinations.
Ethical statement
This article does not contain any studies with human participants or
animals.
Conflict of interest statement
All authors declare that there is no conflict of interest to publish
entire body of this article.
Acknowledgements
This work was subsidized by Faculty of Technology and
Environment, Prince of Songkla University, Phuket campus. Authors
would also like to thank Assoc. Prof. Dr. Seppo Karrila and the Research
and Development Office, Prince of Songkla University for language
polishing.
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