LWT - Food Science and Technology 123 (2020) 109068

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LWT - Food Science and Technology

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A novel method for detection of lipid oxidation in edible oil T

a a b,∗ a
Meizhen Xie , Xiyu Dong , Yan Yu , Leqi Cui
a
School of Food Equipment Engineering and Science, Xi'an Jiaotong University, Xi'an, 710049, China
b
School of Public Health, Xi'an Jiaotong University, Shaanxi, 710049, China

A R T I C LE I N FO A B S T R A C T

Keywords: Lipid oxidation is a major food safety issue for edible oils. The objective of this study was to develop a simple
Lipid oxidation method for detecting lipid oxidation through the color change of a composite film. Schiff's reagent which can
Aldehyde form color compound with lipid oxidation products—aldehydes was incorporated into polyvinyl alcohol (PVA)
Colorimetric response to form the composite film. The effects of PVA molecular weight (Mw), Schiff's reagent/PVA ratio and glycerol
Film
content on the response of the composite film to aldehyde were investigated. Sunflower seed oil with various
Schiff's reagent
extent of oxidation was used to evaluate the effectiveness of the composite film for lipid oxidation detection. The
results showed that the composite film formed with lower PVA Mw turned from colorless to purple upon al-
dehyde exposure. Higher Schiff's reagent/PVA ratio and glycerol content led to shorter response time and larger
color change after aldehyde exposure. The color change of the composite film was linearly (R2 = 0.96) related to
aldehyde concentration. The oxidation status of four sunflower seed oil samples out of five were correctly
identified by the composite film. Although the accuracy of this method was 80% in this study, it still shows great
potential for lipid oxidation detection application.

1. Introduction Sikorska, 2015), nuclear magnetic resonance (Merkx, Hong, Ermacora,

Lipid oxidation is a major food safety issue, especially for edible oils.
Oxidation of edible oils not only decreases the sensory properties of oil
products, it also produces compounds such as hydroperoxides and al-
dehydes that can cause health problems for consumers (Kubow, 1992).
There are well-established methods for detecting lipid oxidation. The
three mostly applied methods are the iodometric method (AOCS, 2009,
pp. 8–87), the p-anisidine value test (AOCS, 2011, pp. 18–90) and the
thiobarburic acid test (AOCS, 2009, pp. 19–90), all of which are wet-
chemistry-based methods for determination of products of lipid oxida-
tion—hydroperoxides and aldehydes. Other methods developed include
the FOX (ferrous oxidation-xylenol orange) assay (Bou et al., 2008), the
DNPH (dinitrophenylhydrazone) method (Esterbauer & Zollner, 1989),
chemiluminescent method (Szterk & Lewicki, 2010), fluorescent
method (Gatellier et al., 2007) and so on. With the advance in instru-
ment technology, liquid chromatography and gas chromatography
coupled with different detectors have been developed for detailed
analysis of hydroperoxides and aldehydes formed during lipid oxidation
(Bauer-Plank & Steenhorst-Slikkerveer, 2000; Katragadda, Fullana,
Sidhu, & Carbonell-Barrachina, 2010). Other technologies such as
electron spin resonance spectroscopy (Thomsen, Kristensen, & Skibsted,
2000), infrared spectroscopy (Wójcicki, Khmelinskii, Sikorski, &
& Van Duynhoven, 2018) and Raman spectroscopy (Li, Driver, Decker,
& He, 2014) have also been developed to determine lipid oxidation in
foods. Although all these methods can provide reliable results, they can
only be performed in a laboratory setting by trained personals, not by
common consumers.
Providing information on the quality of foods through the color
change of a card or strip such as the pH paper is a simple method that
can be performed by ordinary people. Efforts have been made to de-
velop visual detection methods for different foods. Guo, Zhang, Cai,
Shen, and Zhu (2013) have fabricated a visual screening card for de-
termination of residues of organophosphorus and carbamate pesticides
in fruit juices and vegetables. Liao and Li (2010) developed a dipstick
similar to the pregnancy test stick for visual detection of aflatoxin B1 in
grains. Gao, Huang, and Wu (2018) developed a paper strip containing
gold nanoparticles for the detection of melamine in milk. Peng et al.
(2018) also reported the fabrication of a gold nanoparticle-based strip
for determining bisphenol A in foods. So far, few attempts have been
made to develop the visual detection method for detecting lipid oxi-
dation in edible oil.
The objective of this study was to develop a composite film for vi-
sual detection of lipid oxidation in edible oils, based on the colorimetric
reaction between Schiff's reagent and aldehydes—the products of lipid

∗
Corresponding author. School of Public Health, Xi'an Jiaotong University, Xi'an, 710049, China.
E-mail address: yuyan@mail.xjtu.edu.cn (Y. Yu).

https://doi.org/10.1016/j.lwt.2020.109068
Received 13 September 2019; Received in revised form 6 January 2020; Accepted 18 January 2020
Available online 21 January 2020
0023-6438/ © 2020 Elsevier Ltd. All rights reserved.
M. Xie, et al.
oxidation. Schiff's reagent is a colorless solution comprised of para-
rosaniline hydrochloride and sodium sulfite. The reaction with alde-
hyde would turn the colorless solution into reddish purple or bluish
purple. Schiff's reagent has been used to develop a sensor for detecting
the environmental contaminant—formaldehyde (Maruo, Nakamura,
Uchiyama, Higuchi, & Izumi, 2008), it has also been used to fabricate a
sensor for monitoring the shell-life of milk based on its reaction with
the volatile compounds produced by spoilage bacteria (Ziyaina, Rasco,
Coffey, Ünlü, & Sablani, 2019). In this study, Schiff's reagent was in-
corporated into a film-forming polymer — polyvinyl alcohol (PVA).
PVA is available in different molecular weights (Mw), which makes Mw
a factor that may influence the performance of PVA as the carrier of
Schiff's reagent. In order to obtain a responsive and sensitive composite
film, Schiff's reagent as a reactant should be incorporated into the film
as much as possible. The accessibility of aldehyde to the incorporated
Schiff's reagent should be maximized. This can be achieved by adding
plasticizers which have been reported to modify the permeability of
polymer films (Sothornvit & Krochta, 2000). In the study, the effects of
PVA Mw, Schiff's reagent/PVA ratio and amount of glycerol as the
plasticizer on the response of the Schiff/PVA composite film to alde-
hyde were investigated and optimized using single-factor experiment
design. The effectiveness of the Schiff/PVA composite film for detecting
lipid oxidation was assessed using oxidized sunflower seed oil.
2. Materials and methods
2.1. Materials
Three different polyvinyl alcohol (molecular weight: 9000–10000,
31000-50000, and 85000-124000), Schiff's reagent (containing para-
rosaniline, hydrochloride and sodium sulfite), and hexanal were ob-
tained from Sigma. Glycerol (HPLC grade) was purchased from Aladdin,
China.
2.2. Preparation of Schiff/PVA composite film
PVA (1.5 g) of different Mw (according to Section 2.1) was mixed
with 30 mL distilled water in a 50 mL Erlenmeyer flask, the flask was
tightly sealed and placed in a water bath with magnetic stirring at 90 °C
untill completely dissolved. The solution is then cooled to room tem-
perature, yielding a 0.05 g/mL PVA solution. Schiff's reagent was mixed
with the PVA solution at ratios from 1:10 to 4:10 (v:v), and glycerol was
added to the mixture at various amount (0–1%) based on the volume of
PVA solution (see Table 1 for detailed information). An aliquot (1 mL)
of the mixture solution was then pipetted into a plastic plate
(44 × 44 mm), and placed in an oven at 35 °C for 5 h to remove water,
and form the Schiff/PVA composite film. The composite films were
stored in sealed containers in an atmosphere of air at room temperature
before analysis.
2.3. Characterization of Schiff/PVA composite film
The morphologies of the Schiff/PVA composite films were examined
using Scanning Electron Microscope (SEM) (MAIA 3 LMH, TESCAN,
USA). The FT-IR spectra of the films were recorded by attenuated total
reflection (ATR) adjunct of Nicolet IS50 spectrometer (Thermo Fisher
Scientific Co., USA) over the frequency range from 650 to 4000 cm −1.
2.4. Response of Schiff/PVA composite film to aldehyde
Hexanal was used as the model aldehyde. The Schiff/PVA composite
film was placed in a 1 L transparent glass bottle at room temperature,
20 μL of hexanal was injected into the bottle and vaporized. The bottle
was sealed with parafilm immediately. The color change of the com-
posite film after hexanal exposure was detected with naked eyes for
every 5 min, and the time required for the visibly detectable color
2
LWT - Food Science and Technology 123 (2020) 109068
change was recorded as the response time of the film to hexanal. The
composite film was exposed to hexanal for up to 24 h. A control ex-
periment was carried out by placing the composite film in an empty 1 L
transparent glass bottle at room temperature for 24 h. A spectro-
photometer (3nh, Model YS3010, Shenzhen 3nh Technology Co., Ltd,
China) was used to determine the color parameters L* (lightness), a*
(red-green) and b* (yellow-blue) of the Schiff/PVA composite film be-
fore and after hexanal exposure, and the color change ΔE was calcu-
lated by Equation (1) according to CIELab:
ΔE = (ΔL∗)2 + (Δa∗)2 + (Δb∗)2 (1)
Where Δ L = L* – L0*, Δ a = a* – a0*, Δ b = b* – b0*. L0*, a0* and b0*
are the color parameters of the composite film before hexanal exposure;
L*, a* and b* are the color parameters of the film after hexanal or air
(control) exposure.
2.5. Optimization of Schiff/PVA composite film
Based on the response of the Schiff/PVA composite film to aldehyde,
PVA Mw, Schiff/PVA ratio and glycerol content were optimized se-
quentially according to Table 1, to obtain the optimal Schiff/PVA
composite film.
2.6. Relationship between the color change of Schiff/PVA composite film
and aldehyde concentration
The composite film with the optimal PVA Mw, Schiff/PVA ratio and
glycerol content was used to investigate the relationship between the
color change of the film and aldehyde concentration. The films were
placed into 1 L transparent glass bottles at room temperature, various
amounts (0, 2, 4, 6, 8, 10 and 20 μL) of hexanal was injected into the
bottles to obtain hexanal concentrations of 0, 14.63, 29.27, 43.90,
58.54, 73.17 and 146.34 μmol/L. After 24 h, the color change of the
films was determined.
2.7. Evaluation of the effectiveness of Schiff/PVA composite film for
detecting lipid oxidation
2.7.1. Oxidation of sunflower seed oil
An aliquot (2 L) of a commercial sunflower seed oil (Jinlongyu,
Yihai Kerry, Shanghai, China) was placed in a 3 L glass bottle
(310 × 162 mm, H x D) and heated at 70 °C in an forced-air convection
oven. An aliquot of the oil (150 mL) was removed from the bottle on
day 0, 6, 15, 31 and 40, and stored under −20 °C immediately for
further analysis.
2.7.2. Determination of oxidation status of sunflower seed oil
Oxidation status of the sunflower seed oil samples were determined
using the iodometric method according to the AOCS official method Cd
8–87 (AOCS, 2009, pp. 8–87), which measures the peroxide values (PV)
of the oil samples. The acid values (AV) of the samples were also de-
termined using the AOCS official method Cd 3d-63 (AOCS, 2009, pp.
3d-63).
2.7.3. GC-MS analysis of headspace hexanal concentration of sunflower
seed oil
A Gas Chromatography-Mass Spectroscopy system (Trace GC Ultra,
ISQ Single Quadrupole MS, Thermo Scientific, Interscience, Louvain-la-
Neuve, Belgium) coupled to a PAL combi-xt autosampler (CTC,
Zwingen, Switzerland) was used for the analysis. An aliquot of the
sunflower seed oil samples (2 g) was weighed into a glass vial (20 mL)
with a screw cap and septum. The vial was immediately sealed, and
kept at room temperature (about 25 °C) overnight to allow for the
evaporation of hexanal (and other volatiles) into the head space. Before
analysis, the vial was agitated for 10 min at 35 °C. 1 mL of the
 M. Xie, et al.

Table 1
Effects of PVA Mw, Schiff/PVA ratio and glycerol content on the response of Schiff/PVA composite film to hexanal exposure.
PVA Mw Schiff/PVA ratio (v/v) Glycerol content (%) L0* a0* b0* Exposure Response time (min) L* a* b* ΔE

9000–10000 1:10 0 96.29 ± 0.09 1.35 ± 0.02 −7.39 ± 0.05 Air (Control) – 95.84 ± 0.32 1.51 ± 0.07 −7.61 ± 0.24 0.53 ± 0.45
Hexanal 180 72.83 ± 5.90 9.69 ± 3.59 −20.98 ± 5.71 23.90 ± 8.28
31000-50000 89.16 ± 0.12 1.72 ± 0.01 −7.90 ± 0.06 Air (Control) – 89.49 ± 0.35 1.94 ± 0.27 −7.97 ± 0.16 0.48 ± 0.39
Hexanal – 89.35 ± 0.29 1.86 ± 0.03 −8.24 ± 0.07 0.47 ± 0.38
85000-124000 89.23 ± 0.06 1.68 ± 0.04 −7.81 ± 0.07 Air (Control) – 89.46 ± 0.28 1.8 ± 0.16 −7.84 ± 0.05 0.30 ± 0.37
Hexanal – 88.39 ± 0.83 2.41 ± 0.48 −9.22 ± 0.73 1.37 ± 1.06

9000–10000 1:10 0 96.29 ± 0.09 1.35 ± 0.02 −7.39 ± 0.05 Air (Control) – 95.84 ± 0.32 1.51 ± 0.07 −7.61 ± 0.24 0.53 ± 0.45
Hexanal 180 72.83 ± 1.90 9.69 ± 1.59 −20.98 ± 1.71 23.90 ± 4.08
2:10 90.61 ± 0.03 1.40 ± 0.03 −7.31 ± 0.12 Air (Control) – 90.66 ± 0.08 1.59 ± 0.03 −7.24 ± 0.08 0.24 ± 0.16
Hexanal 125 60.55 ± 0.09 6.03 ± 0.04 −33.6 ± 2.29 40.07 ± 2.96
3:10 90.29 ± 0.01 1.69 ± 0.03 −6.48 ± 0.09 Air (Control) – 90.32 ± 0.29 1.66 ± 0.04 −6.77 ± 0.21 0.41 ± 0.40

3
Hexanal 115 61.25 ± 0.53 3.43 ± 0.12 −30.69 ± 1.30 37.87 ± 1.76
4:10 90.22 ± 0.03 1.58 ± 0.02 −6.35 ± 0.04 Air (Control) – 90.06 ± 0.10 1.78 ± 0.05 −6.48 ± 0.08 0.31 ± 0.19
Hexanal 95 64.89 ± 2.11 4.99 ± 0.53 −26.2 ± 1.42 32.69 ± 3.96

9000–10000 2:10 0 90.61 ± 0.03 1.40 ± 0.03 −7.31 ± 0.12 Air (Control) – 90.66 ± 0.08 1.59 ± 0.03 −7.24 ± 0.08 0.24 ± 0.16
Hexanal 125 60.55 ± 0.09 6.03 ± 0.04 −33.6 ± 1.29 40.07 ± 2.36
0.33 90.3 ± 0.08 1.42 ± 0.03 −7.08 ± 0.07 Air (Control) – 89.79 ± 0.19 1.62 ± 0.10 −7.24 ± 0.21 0.6 ± 0.22
Hexanal 80 62.47 ± 2.14 5.53 ± 0.53 −31.7 ± 1.29 37.39 ± 3.37
0.67 89.86 ± 0.09 1.38 ± 0.03 −7.21 ± 0.15 Air (Control) – 88.85 ± 0.12 1.46 ± 0.07 −6.89 ± 0.31 1.10 ± 0.53
Hexanal 55 56.34 ± 2.91 13.19 ± 1.87 −39.47 ± 2.97 48.01 ± 4.48
1 89.42 ± 0.03 1.09 ± 0.02 −7.53 ± 0.04 Air (Control) – 89.01 ± 0.32 0.95 ± 0.14 −6.75 ± 0.28 0.92 ± 0.59
Hexanal 55 51.99 ± 1.18 17.76 ± 1.05 −43.46 ± 0.57 54.53 ± 1.82

Values are means ± SD (n = 3). L0*, a0* and b0* are the color parameters of the film before exposure; L*, a* and b* are the color parameters of the film after hexanal/air exposure. ΔE is the color change of the film
before and after hexanal/air exposure.
― not applicable.
LWT - Food Science and Technology 123 (2020) 109068
M. Xie, et al. LWT - Food Science and Technology 123 (2020) 109068

Fig. 1. SEM micrographs of Schiff/PVA composite

films. A, B and C are films with Mw 9000–10000,
31000–50000, and 85000–124000, respectively
(Schiff/PVA ratio of 1:10, glycerol content of 0%);
D, E and F are films with Schiff/PVA ratios of 2:10,
3:10 and 4:10, respectively (PVA Mw 9000–10000,
glycerol content of 0%); G, H and I are films with
glycerol contents of 0.33, 0.67 and 1%, respectively
(PVA Mw 9000–10000, Schiff/PVA ratio of 2:10).

Fig. 2. IR spectra of Schiff/PVA composite films. A, B and C are films with Mw 85000–124000, 31000–50000, and 9000–10000, respectively (Schiff/PVA ratio of
1:10, glycerol content of 0%); D, E and F are films with Schiff/PVA ratios of 2:10, 3:10 and 4:10, respectively (PVA Mw 9000–10000, glycerol content of 0%); G, H
and I are films with glycerol contents of 0.33, 0.67 and 1%, respectively (PVA Mw 9000–10000, Schiff/PVA ratio of 2:10).

headspace gas was taken using a Head Space syringe (2.5 mL, CTC min. The mass spectrometer was operated in the electron impact (EI)
Analytics, MSH 02-00B, Zwingen, Switzerland) and injected into an ionization mode at 70 eV, and the data was acquired in the mass range
Equity DB-1 column (30 m × 0.32 mm × 1 μm). The GC separation of 30–400 amu. A standard curve was established with 0–83 μmol/L of
was carried out at 65 °C for 10 min, with a helium flow rate of 15 mL/ hexanal to quantify hexanal concentration in headspace of sunflower

4
M. Xie, et al.
Fig. 3. Schiff/PVA composite films before and after hexanal (146.34 μmol/L)
exposure. A: Composite films with different PVA Mw (Schiff/PVA ratio of 1:10,
glycerol content of 0%); B: Composite films with different Schiff/PVA ratios
(PVA Mw 9000–10000, glycerol content of 0%); C: Composite films with dif-
ferent glycerol contents (PVA Mw 9000–10000, Schiff/PVA ratio of 2:10).
seed oils.
2.7.4. Response of Schiff/PVA composite film to oxidized sunflower seed oil
As described above, 2 g of oxidized sunflower seed oil was weighed
into a 20 mL glass vial, sealed and kept at room temperature overnight.
The composite film was placed into in the vial and positioned above the
oil level. The vial was sealed. After 24 h, the color change of the film
was measured. A control experiment was performed by placing the film
into an empty vial for 24 h.
5
LWT - Food Science and Technology 123 (2020) 109068
3. Results and discussion
3.1. Characteristics of Schiff/PVA composite films
Fig. 1 shows the morphologies of the Schiff/PVA composite films.
Although the films appeared colorless and transparent to human eyes
(Fig. 3A, first row), under SEM, particles of about 10 μm were observed
scattered in the films with PVA Mw 9000–10000 and 31000-5000
(Fig. 1A and B), which were probably the crystals of the Schiff's reagent
component. Net-like structures up to 100 μm were observed in the film
with PVA Mw 85000-124000 (Fig. 1C), which seemed to be the frag-
ments of the polymer itself. Schiff/PVA ratio of 1:10 and 2:10 resulted
in colorless and transparent films, but Schiff/PVA ratio of 3:10 and 4:10
resulted in colorless and opaque films (Fig. 3B, first row). The opa-
queness was due to a layer of fine crystals observed on the surface of the
films. Apparently, increasing the amount of Schiff's reagent resulted in
the precipitation and crystallization of its components on the surface of
the films. Under SEM, more particles were observed in the film with
Schiff/PVA ratio of 2:10 (Fig. 1D) than the one with Schiff/PVA ratio of
1:10 (Fig. 1A); the films with Schiff/PVA ratios of 3:10 and 4:10 were
full of fine particles (Fig. 1E and F), which was consistent with the
visual observation. The films with different glycerol contents were
colorless and transparent (Fig. 3C, first row). Under SEM, less particles
were observed in the films with the increase in glycerol content
(Fig. 1D, G, H and I). The IR spectra of all the films were similar (Fig. 2).
The absorptions peaks at 3396 (O–H stretching), 2918 (C–H stretching),
1492 (C–H bending) and 1451 cm−1 (C–H bending) were associated
with PVA (Krimm, Liang, & Sutherland, 1956). The small absorption
peak at 1636 cm−1 was due to N–H bending (Ravi, Shanmugam,
Subbaiah, Prasad, & Reddy, 2017) in pararosaniline, the component of
Schiff's reagent. The ATR-FTIR spectra confirm the existence of Schiff's
reagent in the films, more specifically at the surface of the films, which
would facilitate the reaction of Schiff's reagent with aldehydes.
3.2. Response of Schiff/PVA composite film to aldehyde affected by PVA
Mw, Schiff/PVA ratio and glycerol content
The effect of PVA Mw on the response of the composite film to al-
dehyde was evaluated at a Schiff/PVA ratio of 1:10 (v/v) and a glycerol
content of 0% (Table 1). When exposed to hexanal, the composite film
prepared with PVA Mw 9000–10000 (Schiff/PVA 9000) turned from
colorless to a visually noticeable purple color (Fig. 3A, second row). The
color of the composite films with PVA Mw 31000-50000 (Schiff/PVA
31000) and 85000-124000 (Schiff/PVA 85000) remained unchanged
after 24 h hexanal exposure (Fig. 3A, second row). The response time
for the color change of Schiff/PVA 9000 upon hexanal exposure was
180 min (Table 1). In the control experiment (exposure in air), all the
three composite films (picture not shown) remained colorless and were
visually similar to the ones before exposure (Fig. 3A, first rows). The
color parameters of the composite films before and after hexanal/air
exposure were shown in Table 1. The L* value describes the brightness
of a color, and ranges from 0 to 100, where 0 means black and 100
means the maximum brightness visible to human eyes. The a* and b*
values describe a color in the green→red and blue→yellow fields, re-
spectively, and both range from −128 to +127 (Mokrzycki & Tatol,
2011). It can be seen from Table 1 that the color change ΔE of Schiff/
PVA 9000, Schiff/PVA 31000 and Schiff/PVA 85000 in the control was
lower than 1. After hexanal exposure, ΔE of Schiff/PVA 9000 was
higher than 5, while ΔE of Schiff/PVA 31000 and Schiff/PVA 85000
was lower than 2. Based on experimentally verified statistics, when ΔE
is lower than 1, observers would not notice the difference between two
colors; when ΔE is between 1 and 2, the color difference can only be
distinguished by experienced observer; when ΔE is higher than 5, ob-
servers would notice the difference between two colors (Mokrzycki and
Tatol, 2011). Our result agrees well with this rule. Since PVA Mw
9000–10000 resulted in a composite film with short response time and
M. Xie, et al. LWT - Food Science and Technology 123 (2020) 109068

Fig. 4. Response of Schiff/PVA film (PVA Mw 9000–10000, Schiff/PVA ratio of 2:10, glycerol content of 1%) to various hexanal concentrations. A: pictures of Schiff/
PVA film after exposed to various hexanal concentrations; B: relationship between hexanal concentration and color change ΔE of Schiff/PVA film. (For interpretation
of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Table 2
Color response of Schiff/PVA composite film to various hexanal concentrations.
Hexanal concentration (μmol/L) L* a* b* ΔE

Before exposure 89.42 ± 0.03 a 1.09 ± 0.02 a −7.53 ± 0.04 a –

0 (air) 89.01 ± 0.32 a 0.95 ± 0.14 a −6.75 ± 0.28 b 0.92 ± 0.59
14.63 88.83 ± 0.45 a 2.07 ± 0.21 b −9.04 ± 0.55 c 1.89 ± 0.84
29.27 85.55 ± 0.42 b 4.26 ± 0.78 c −12.86 ± 0.50 d 7.31 ± 1.05
43.90 82.81 ± 1.36 c 5.41 ± 0.93 cd −16.02 ± 1.21 e 10.59 ± 1.88
58.54 77.87 ± 0.76 d 5.90 ± 0.71 d −20.59 ± 0.27 f 18.08 ± 1.26
73.17 73.9 ± 0.36 e 9.43 ± 0.58 e −25.35 ± 0.38 g 25.06 ± 0.89
146.34 51.99 ± 1.18 f 17.76 ± 1.05 f −43.46 ± 0.57 h 54.53 ± 1.82

The Schiff/PVA composite film was prepared with PVA Mw 9000–10000, Schiff/PVA ratio of 2:10 and glycerol content of 1%.
Values are means ± SD (n = 3). L*, a* and b* are the color parameters of the film after hexanal exposure. Means in the same column with different letters are
significantly different from each other (P < 0.05).
― not applicable.

large ΔE, it was used in the following experiments to prepare composite
films.
The effect of Schiff/PVA ratio was investigated at a glycerol content
of 0%. All the composite films turned purple after exposure to hexanal
(Fig. 3B), and remained colorless in the control experiment (picture not
shown). The response time for the composite film to turn purple de-
creased from 180 min to 95 min as Schiff/PVA ratio increased from
1:10 to 4:10 (Table 1). This result suggests that more Schiff's reagent is
incorporated into the composite film, the faster the color response to
hexanal. All the composite films in the control had ΔE less than 1, and
the films exposed to hexanal had ΔE larger than 5. After hexanal
exposure, ΔE increased when Schiff/PVA ratio increased from 1:10 to
2:10, and then decreased as Schiff/PVA ratio increased to 4:10
(Table 1). The decrease in ΔE with the increase in Schiff/PVA ratio to
4:10 was possibly due to the deposition of the crystals on the film
surface. Although Schiff/PVA ratio of 3:10 and 4:10 resulted in shorter
response time, the appearance of crystals on the film surface would
affect the application of the films. For this reason, Schiff/PVA ratio of
2:10 was regarded as the optimal ratio, and used in the following ex-
periment.
It has been reported that addition of plasticizers can modify the gas
permeability of polymer films (Sothornvit & Krochta, 2000). Glycerol

6
M. Xie, et al. LWT - Food Science and Technology 123 (2020) 109068

was used as the plasticizer in this study. Four different amounts (based
on the volume of PVA solution) (0, 0.33, 0.67 and 1%) of glycerol were
used to prepare the Schiff/PVA composite film. All the composite films
Equivalent headspace hexanal

Values are Means ± SD (n = 3). L*, a* and b* are the color parameters of the film after exposure to.Means in the same column with different letters are significantly different from each other (P < 0.05).
remained visually unchanged in the control (picture not shown), and
turned purple after hexanal exposure (Fig. 3C). The response time of the
concetration (μmol/L)

composite films shortened from 125 min to 55 min as the glycerol

amount increased from 0 to 1% (Table 1). This may be due to the fact
that glycerol is able to weaken the intermolecular interaction of PVA
(Sothornvit & Krochta, 2000), resulting in composite films with less
111.36
12.45
98.52
5.53
7.03

dense structure, which makes it easier for hexanal to penetrate the

–

structure and react with the incorporated Schiff's reagent. The ΔE for
34.18 ± 0.73
38.63 ± 1.21

the composite films in the control was less than 1, which is consistent
0.92 ± 0.59
1.92 ± 0.78
2.44 ± 1.41
4.32 ± 0.82

with the observation. For the films exposed to hexanal, the ΔE was
larger than 5, and generally increased as glycerol content increased
(Table 1). The results are in good correlation with the SEM observation
ΔE

(Fig. 1D, G, H and I), where the structure of the composite films became
−24.98 ± 0.23 c
−26.72 ± 0.41 c
−7.41 ± 1.07 ab
b

−8.62 ± 0.49 b
a

more and more uniform with less amount of particles as the glycerol
−6.73 ± 0.4 b
−7.53 ± 0.04

−6.75 ± 0.28

amount increased.
Based on the results, the Schiff/PVA composite film prepared with
PVA Mw 9000–10000, Schiff/PVA ratio of 2:10 and glycerol content of
1% resulted in a shorter response time and largest color change ΔE after
b*

hexanal exposure, and also exhibited the most uniform structure

24.35 ± 0.76 d
20.54 ± 0.37 c
1.14 ± 0.38 b
a

0.04 ± 0.28 a
0.48 ± 0.89 a

(Fig. 1I), it was regarded as the optimal film for lipid oxidation detec-
1.09 ± 0.02

0.95 ± 0.14

tion, and used in the following experiment.

3.3. Effect of hexanal concentration on color change of Schiff/PVA

a*

composite film
a

The Schiff/PVA composite film was prepared with PVA Mw 9000–10000, Schiff/PVA ratio of 2:10 and glycerol content of 1%.
90.32 ± −0.22
90.44 ± −0.55

70.67 ± 0.81 b
a

89.09 ± 0.29 a
72.7 ± 0.24 b
89.42 ± 0.03

89.01 ± 0.32

Fig. 4A shows the pictures of the Schiff/PVA composite film after

Oxidation status, headspace hexanal concentration, and color response of sunflower seed oil to Schiff/PVA composite film.

exposed to various concentrations of hexanal. As the hexanal con-

centration increased from 0 (control) to 146.34 μmol/L, the color of the
L*

film changed from colorless to pale purple, and then to purple as ob-
served in Fig. 4A. This change in the color visually observed was con-
sistent with the color parameters (Table 2). As hexanal concentration
increased, the L* value representing the brightness decreased, the a*
concetration (μmol/L)

value became more positive meaning a shift towards red, and the b*
Headspace hexanal

value in general became more negative meaning a shift towards blue

≤ 0.11 ± 0.04
≤ 0.26 ± 0.05

(Table 2). As a result, the film became purple, and ΔE became larger
with the increase in hexanal concentration. A linear relationship
(R2 = 0.96) was observed between hexanal concentration and ΔE
n.d
n.d
n.d

(Fig. 4B). This result shows that the composite film not only can be used
–

for qualitative analysis, with the aid of an instrument, it can also be

Acid value (mg KOH/

used for quantitative analysis.

b
a

c
0.08
0.04
0.09
0.04
0.05

3.4. Effectiveness of Schiff/PVA composite film for lipid oxidation detection

±
±
±
±
±

A commercial sunflower seed oil was subjected to accelerated oxi-

0.34
0.31
0.33
0.64
1.17
g)

dation process at 70 °C for various days. The two essential quality cri-
–

teria of edible oils – PV and AV related to lipid oxidation were de-

termined and presented in Table 3. As the time of oxidation prolonged,
Peroxide value(milliequivalents of

both PV and AV increased. According to Codex Alimentarius Commis-

sion (CAC), the limits for PV and AV of edible oils are 10 milli-
equivalents of active oxygen/kg oil and 0.6 mg KOH/kg, respectively. It
can be seen from Table 3 that after 15 days of oxidation, the PV or PV/
active oxygen/Kg)

AV of the oil was over the value limit, which means the oil has been
119.24 ± 0.69 e
90.92 ± 0.28 d
36.36 ± 0.67 c
8.62 ± 0.03 b
1.14 ± 0.06 a

oxidized and not suitable for human consumption. Since hexanal is one
of the major aldehydes formed during oxidation of sunflower seed oil
(Guillén, Cabo, Ibargoitia, & Ruiz, 2005) and used as the model alde-
hyde in this study, its presence in the headspace gas of the oxidized
–

sunflower seed oil samples was determined. However, the headspace

hexanal concentration of the oil samples was too low for detection at
― not applicable.
Film in Control
Oxidation days

room temperature (25 °C), which is the condition expected for the
exposure
Film Before

composite film to function, the headspace hexanal concentration was

determined at 35 °C instead. For sunflower seed oil oxidized for less
Table 3

than 31 days, it was unable to detect the headspace hexanal con-

15
31
40
0
6

centration with the method used. After 31 days of oxidation, the

7
M. Xie, et al.
Fig. 5. Pictures of Schiff/PVA film (PVA Mw 9000–10000, Schiff/PVA ratio of 2:10, glycerol content of 1%) after exposed to headspace gas of sunflower seed oils
oxidized for different days.
headspace hexanal concentration was detectable and reached a max-
imum of no more than 0.26 μmol/L.
As shown in Fig. 5, for sunflower seed oil oxidized for 0 and 6 days,
the Schiff/PVA composite film correctly indicated their oxidation status
and edibility by remaining colorless. For sunflower seed oil oxidized for
15 days, the composite film failed to indicate its oxidation status by
staying colorless. For the oil oxidized for 31 and 40 days, the film was
able to correctly reveal the oxidation status by turning into a reddish
purple color. It is noticed that the headspace hexanal concentration of
sunflower seed oil oxidized for 31 and 40 days (Table 3) was much
lower than the hexanal concentration required to induce the color re-
sponse on the composite film (Table 2). This was not surprising, since
other volatile aldehydes formed during oxidation can also react with
the incorporated Schiff's reagent. As the oxidation of sunflower seed oil
proceeded, the L* value of the film became lower, the a* value became
more positive, and the b* value became more negative. This change in
these three color parameters of the film with increased oxidation was
similar to that obtained with increased hexanal concentration (Table 2).
Consequently, the ΔE increased with the increase in the extent of sun-
flower seed oil oxidation (Table 3). In general, lipid oxidation detection
by the composite film developed resulted in a detection accuracy of
80% in this study.
It is noticeable that the headspace gas of oxidized sunflower seed oil
resulted in a reddish purple color (Fig. 5) different from the purple color
observed in Figs. 3 and 4. This result was not unexpected. The color
observed in Figs. 3 and 4 was caused by the reaction between Schiff's
reagent and hexanal. In the headspace gas of the oxidized sunflower
seed oil, besides hexanal, many other aldehydes (pentanal, nonanal,
and 2, 4-decadienal, etc.) were also present (Guillén et al., 2005) and
reacted with the incorporated Schiff's reagent. As a result, the compo-
site film exposed to the headspace gas of the oxidized sunflower seed oil
exhibited a different color from that exposed to hexanal. In order to
provide a general information on the total aldehyde concentration in
the headspace gas of the oxidized sunflower seed oil samples, the
equivalent headspace hexanal concentration of the oil was calculated
(Table 3), using the linear model established between hexanal con-
centration and ΔE (Fig. 4B) in this study.
4. Conclusions
The colorimetric response of the Schiff/PVA composite film to al-
dehyde was influenced by PVA Mw, Schiff/PVA ratio and glycerol
content. Lower PVA Mw, higher Schiff/PVA ratio and glycerol content
resulted in a Schiff/PVA composite film with shorter response time and
larger color change ΔE upon aldehyde exposure. The optimal Schiff/
PVA composite film was prepared with PVA Mw 9000–10000, Schiff/
PVA ratio of 2: 10 and glycerol content of 1%. The detection accuracy of
the composite film for indicating the oxidative status and edibility of
sunflower seed oil was 80% in this study. Apparently, a detection ac-
curacy of 80% is not satisfactory considering the detrimental impact of
lipid oxidation on human health. However, based on our results, the
developed composite film was very effective in indicating highly oxi-
dized edible oils, which means this film could at least be used to warn
8
LWT - Food Science and Technology 123 (2020) 109068
consumers against edible oils with high extent of lipid oxidation. Still,
the improvement on the detection accuracy of the Schiff/PVA compo-
site film for lipid oxidation determination is required.
Author Statement
Meizhen Xie: Conceptualization, Methodology, Investigation,
Writing- Original draft preparation, Writing- Reviewing and Editing.
Xiyu Dong: Investigation, Formal analysis, Data curation. Yan Yu:
Resources, Supervision. Leqi Cui: Validation
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
This work was supported by the National Natural Science
Foundation of China (Grant numbers 31701523, 21706222).
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