Food Chemistry 293 (2019) 340–347

Contents lists available at ScienceDirect

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

The synthesis, adsorption mechanism and application of polyethyleneimine T

functionalized magnetic nanoparticles for the analysis of synthetic colorants
in candies and beverages
⁎ ⁎
Hui Chena, Xiaojuan Dengb, Guosheng Dingb, , Yuanyuan Qiaoa,
a
Institute of Applied Chemistry, College of Chemistry, Nankai University, 94 Weijin Road, Tianjin 300071, PR China
b
Analysis Center, Tianjin University, 92 Weijin Road, Tianjin 300072, PR China

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

Keywords: A green and simple method was designed to synthesize polyethyleneimine (PEI)-modified magnetic nano-
Polyethyleneimine particles (MNPs), which were later used as adsorbents in magnetic solid phase extraction (MSPE) process. A new
Polydopamine MSPE-HPLC method was then established for the simultaneous determination of four representative synthetic
Magnetic solid phase extraction colorants (Amaranth, Ponceau 4R, Sunset yellow and Allure red) in food samples. Several important factors, such
Colorants
as the pH of the sample solution, the amount of adsorbent, the adsorption time, and the type of the eluent were
HPLC
investigated in detail. Under the optimized experimental conditions, the four colorants were measured with good
linearity, detection limit and quantification limit. Adsorption kinetics and isotherms were also investigated to
elucidate the adsorption mechanism. The detection of four colorants in candy, jelly and carbonated drink proved
that the established MSPE-HPLC method was simple and effective and can be used for the analysis of colorants in
real samples.

1. Introduction
Color is always the first sensory quality by which foods are judged.
To make food more attractive and appetizing, natural colorants have
been used for thousands of years. Owning to higher tinctorial strength,
stability and lower price, some synthetic colorants have been ex-
tensively utilized in food industry since the 1880s (Knill & Kennedy,
1996). However, some synthetic colorants have been found to be po-
tentially toxic and their use is therefore prohibited (Chen, Mou, Hou,
Riviello, & Ni, 1998).
Many chromatographic techniques have been used for the simulta-
neous determination of various synthetic colorants, such as capillary
electrophoresis (CE) (Huang, Shih, & Chen, 2002), thin layer chroma-
tography (TLC) (Soponar, Moţ, & Sârbu, 2008), HPLC-UV–Vis (Ertaş,
Özer, & Alasalvar, 2007), HPLC-DAD (Alves, Brum, Ecbde, & Pereira
Netto, 2008; Kiseleva, Pimenova, & Eller, 2003; Minioti, Sakellariou, &
Thomaidis, 2007) and HPLC-MS (Gosetti et al., 2004). In order to se-
parate colorants from complex matrixes, and to obtain higher detection
sensitivity at the same time, sample pretreatment process is indis-
pensable. Among different sample pretreatment processes widely used
nowadays, magnetic solid phase extraction (MSPE) is the preferred one
because it can realize phase separation only by applying an external
magnetic field (no need of filtration or centrifugation). In comparison
with conventional solid phase extraction (SPE) process, it is also time-
saving because of the avoidance of the problem of blockage, as well as
the step of sample loading.
In recent years, Fe3O4 based magnetic NPs (MNPs) are gaining ex-
tensive attention in the field of MSPE, because of their large surface
area, low cost, and the unique superparamagnetism at room tempera-
ture (Feng, Cao, Ma, Zhu, & Hu, 2012; Long, Xiao, & Cao, 2017). In
order to achieve the analysis of sample components with different
structural characteristics, Fe3O4 NPs are usually modified (or functio-
nalized) with different materials (or functional groups) prior to use to
improve their stability, or to introduce new surface characteristics.
Dopamine (DA) can form a thin, surface-adhered polydopamine
(PDA) coating under mild conditions. As an ideal polymer material,
PDA has attracted multidisciplinary interests in the past decade, due to
its strong adhesion, active surface and ease of preparation (Rodríguez
et al., 2016; Zheng et al., 2017). The strong adhesion of PDA is mainly
due to catechol group it contains, which can form covalent or non-

Abbreviations: MNPs, magnetic nanoparticles; DA, dopamine; PDA, polydopamine; MSPE, magnetic solid-phase extraction; ACN, acetonitrile; HPLC, high perfor-
mance liquid chromatography
⁎
Corresponding authors.
E-mail address: dgs@tju.edu.cn (G. Ding).

https://doi.org/10.1016/j.foodchem.2019.04.111
Received 14 January 2019; Received in revised form 27 April 2019; Accepted 28 April 2019
Available online 29 April 2019
0308-8146/ © 2019 Elsevier Ltd. All rights reserved.
H. Chen, et al.
covalent bond interactions (hydrogen bond, van der waals force and π-
π stacking etc) with the surface of the many substrates, such as gra-
phene oxide (Cheng et al., 2013), mesoporous silica (Jun-Kai, Li-An,
Guang-Hui, & Ping, 2015), and magnetic Fe3O4 (Chai, Wang, Zhang, &
Ding, 2016). Due to the structural characteristics of PDA (many dif-
ferent functional group, such as phenyl, amino, imine and phenol
groups in the structure), PDA-modified MNPs (PDA@Fe3O4) had been
tested for the binding with hazardous compounds, Gram-negative
bacterium (Zafar, Samar, Musharraf, & Lai, 2011), polycyclic aromatic
hydrocarbons (Wang et al., 2013) and colorants (Chai et al., 2016) in
the past several year. Despite the success of these studies, the lack of
binding specificity is an obvious weakness for PDA@Fe3O4.
Polyethyleneimine (PEI) is a water-soluble polymer, which has a
large number of polar amino groups on its macromolecular chains. PEI
can be protonated at pH values lower than 10, so it is a cationic poly-
electrolyte (Gao, Chen, Su, Zhang, & Peng, 2009). There is strong
electrostatic interaction between PEI and compounds (or materials)
which are negatively charged, allowing PEI to be used as adsorbents in
water treatment, as plating solution or dispersing agent in other ap-
plication areas (Zhang et al., 2016). For example, Nayab (Nayab et al.,
2014) modified branched PEI on the surface of mesoporous SiO2 for
adsorption of Alizarin red and Xylenol orange. The results showed that
a large amount of amino groups in PEI molecule can significantly im-
prove the adsorption performance.
In this work, PEI modified MNPs (PEI@PDA@Fe3O4) were prepared
for the first time by a facile and green process, in which very little
organic solvents were used. PDA layer was firstly formed on the surface
of Fe3O4 NPs, then PEI was further modified using PDA layer as the
platform for secondary reaction. Four food colorants (all containing
aromatic rings and sulfonic acid groups in molecular structures) were
selected to study the adsorption properties of the materials. HPLC
analysis was used to examine some parameters affecting the extraction
efficiency, mainly including the pH value of the sample solution, the
amount of adsorbent, extraction time and desorption conditions. The
experimental results show that the established MSPE-HPLC method for
the determination of four colorants in food samples has the advantages
of simple operation, good analytical reproducibility and high sensi-
tivity.
2. Experimental
2.1. Chemicals and reagents
Ferric chloride hexahydrate (FeC13·6H2O), sodium acetate anhy-
drous (NaAc), polyethylene glycol 6000 (PEG 6000) and ethylene
glycol (EG) were all analytical grade and obtained from Tianjin
Guangfu Fine Chemical Research Institute (Tianjin, China).
Trihydroxymethylaminomethane hydrochloride (Tris-HCI) was pur-
chased from Tianjin Weiyi Chemical Technology Co., Ltd. (Tianjin,
China). Dopamine hydrochloride and branched polyethyleneimine
(Mw = 10,000) were both purchased from Alfa Aesar (Heysham, UK).
Sodium dihydrogen phosphate, hydrochloric acid and phosphoric
acid were all from Beijing Chemical Reagent Company (Beijing, China).
Ammonia (25%, wt%) with HPLC grade was purchased from Aladdin
Chemistry Co. Ltd. (Shanghai, China). Acetonitrile (ACN) and methanol
(MeOH) were HPLC grade and supplied by Concord Technology Co.,
Ltd. (Tianjin, China). Colorant standards of Amaranth, Ponceau 4R,
Sunset yellow and Allure Red were bought from National information
center (Beijing, China). They were all in the liquid state (in pure water)
as received, with the concentration values of 0.500 mg mL−1 (for sunset
yellow) and 1.00 mg mL−1 (for other three synthetic colorants). In
order to reduce the errors, we used the same batches of reference ma-
terials in the whole experiment. The ultrapure water used in experi-
ments was prepared by a Milli-Q system (Millipore, USA).
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Food Chemistry 293 (2019) 340–347
2.2. Instruments
Agilent 1260 HPLC system (Agilent Technologies, USA), consisting
of a quaternary pump, diode array detector (DAD) and Chemstation
software, was used for the chromatographic analysis. For extraction
experiments, thermostatic water bath oscillator with the model of WE-2
(Tianjin Honor Instrument Co., Ltd.) was used with the setting of
280 rpm. A BF-2000 termovap sample concentrator (BFC, China) was
used to concentrate the eluants.
JEM-2100F transmission electron microscopes (JEOL, Japan),
Nanosem 430 scanning electron microscopy (FEI, USA) installed with
Energy dispersive spectrometry spectrometer (EDX, USA), were em-
ployed to characterize the morphology of the PEI@PDA@Fe3O4 nano-
composites. The analysis of X-ray diffraction (XRD) pattern was per-
formed on the D/MAX-2500 X-ray diffraction (Rigaku, Japan). Zetasizer
Nano ZS (Malvern Instruments, UK) was used for the zeta potential
determination. A 7404 vibrating sample magnetometer purchased from
the Lake Shore Cryotronics Inc. (USA) was used to characterize the
magnetic properties of synthetic materials at the temperature of 298 K
and magnetic field strengths of −2 T–2 T.
2.3. Preparation of PEI@PDA@Fe3O4
Magnetic Fe3O4 NPs were synthesized by a solvothermal method,
similar to that reported by Li et al. (Deng et al., 2005) with slight
modifications. Briefly, 2.7 g FeCl3·6H2O and 7.2 g NaAc were dissolved
in 80 mL EG under ultrasound, and then mixed thoroughly by magnetic
stirring at room temperature for 3 h. After that, 3.0 g PEG 6000 was
added to the above yellow solution. After stirring, the mixture was
transferred to a stainless steel reactor and sealed, and reacted at 200 °C
for 10 h. After cooling to room temperature, the product was washed
three times with anhydrous ethanol and ultrapure water (10 mL per
time) sequentially. To ensure thorough washing, UV–Vis spectro-
photometer with the model of Lambda 750 was used for the mon-
itoring. The product was finally dried under vacuum at 50 °C for 2 h.
PDA@Fe3O4 nanocomposites were formed by the oxidization and
self-polymerization of DA under alkaline conditions. Typically, 180 mg
Fe3O4 NPs were ultrasonically dispersed in 80.0 mL Tris-HCl solution
(10 mM, pH 8.5) at room temperature, and then 180 mg dopamine
hydrochloride in 10.0 mL Tris-HCl solution was added dropwise. After
mechanical stirring for 12 h, the product was collected with a magnet,
washed several times with deionized water (10 mL per time), and then
vacuum dried at 50 °C for 2 h.
For the preparation of PEI@PDA@Fe3O4, 200 mg PDA@Fe3O4 na-
nocomposites prepared in the last step were dispersed in 100 mL Tris-
HCl (10 mM, pH 8.5) solution. PEI aqueous solution (50 mg mL−1) was
slowly added, and the mixed solution was kept in 50 °C water bath for
12 h with mechanical stirring. The resultant materials were finally
collected by a magnet, washed several times with deionized water, and
then dried under vacuum at 50 °C for 2 h. The whole preparation pro-
cedure of PEI@PDA@Fe3O4 NPs is schematically represented in Fig. S1.
2.4. Sample preparation
Stock solutions of four synthetic colorants (0.2 mg mL−1 each) were
prepared in ultrapure water, and then stored at 4 °C prior to use. The
working standard solutions were prepared by successively diluting the
stock solutions with dilute hydrochloric acid. Four food samples, in-
cluding candy, jelly, and carbonated drink, were purchased at the local
retail supermarket. The candy were accurately weighed and crushed,
and then dissolved into ultrapure water; the jelly was heated in water
bath until it became solution, and then accurately weighed; carbonated
drink was directly weighed and degassed by ultrasonication. All sam-
ples were filtered with 0.45 μm filter membrane to remove particulate
matter, and diluted to a certain volume with ultrapure water, and fi-
nally adjusted to pH = 2.0 with 1 M hydrochloric acid.
H. Chen, et al.
2.5. MSPE procedure
In order to extract four synthetic colorants more effectively, several
parameters influencing the MSPE process were optimized by changing
one parameter at a time, while the other parameters remained the
same. In a typical experiment, 30 mg PEI@PDA@Fe3O4 NPs were added
into 100 mL sample aqueous solution, and then shaken on a shaker
(280 rpm) for 10–50 min. After that, a strong magnet was applied and
the supernatant was decanted. Using CH3CN-NH3·H2O (70:30, v/v) as
eluent (1 mL, repeat three times), analytes on PEI@PDA@Fe3O4 were
eluted under ultrasonication. Using a stream of nitrogen, the desorption
solution was evaporated at 55 °C. Ultrapure water (500 μL) was used to
re-dissolve the residue, and the obtained solution was filtered with
0.22 μm filter membrane, prior to injection into HPLC instrument for
the measurement.
2.6. Adsorption mechanism studies
Adsorption kinetics experiments were carried out as follows: a
certain amount of PEI@PDA@Fe3O4 NPs was separately added to a
solution of four colorants (pH = 2.0) at the concentration of
100 mg L−1. The temperature was kept at 25 °C and the adsorption time
was 5 min, 10 min, 15 min, 25 min, 30 min, 60 min, 100 min and
200 min, respectively. The supernatants at different time were taken for
HPLC analysis to determine the concentration of colorants.
To obtain the corresponding adsorption isotherms, PEI@PDA@
Fe3O4 NPs were added into solutions containing different initial con-
centration of four colorant solutions (5–80 mg L−1). The mixtures were
treated with the same procedure as above-mentioned at room tem-
perature. Magnetic separation was performed after adsorption equili-
brium, and the concentration of the colorants in the supernatant was
determined by HPLC analysis.
2.7. HPLC analysis of colorants
The separation was carried out on a Venusil XBP C18 column
(150 × 4.6 mm i.d., 5 μm) under the temperature of 30 °C. By evalu-
ating the wavelength range of 200–700 nm, 515 nm (for Amaranth,
Ponceau 4R and Allure red) and 480 nm (for Sunset yellow) were finally
selected for quantitative analysis. Using 20 mmol L−1 ammonium
acetate (solvent A) and methanol (solvent B) as components, the linear
gradient was optimized as: 0–7 min, 10–34% (B); 7–9 min, 34% (B);
9–12 min, 34–65% (B); 12–12.1 min, 65–10% (B); 12.1–16 min, 10%
(B). The flow rate was 1.0 mL min−1.
3. Results and discussion
3.1. Characterization of PEI@PDA@Fe3O4 NPs
As can be seen from TEM images (Fig. 1), the particles are all
spherical and well dispersed. In comparison with PDA@Fe3O4 NPs, the
surface of PEI@PDA@Fe3O4 NPs becomes rough and the interface be-
tween the organic layer and inorganic core becomes blurred, after the
modification step of branched PEI.
The spectra from energy-dispersive X-ray analysis are shown in Fig.
S2. Compared with the EDX spectrum of PDA@Fe3O4 NPs, the relative
contents of C and N in PEI@PDA@Fe3O4 NPs increased significantly,
indicating the successful bonding of PEI on the surface of PDA@Fe3O4
NPs, mainly through the chemical reaction of amino groups (in PEI)
with the quinone groups (in PDA).
Due to the existence of ionizable functional groups on the surfaces,
the electric properties of the functionalized magnetic NPs would change
with the change of pH of the aqueous solution. In the zeta potential
testing experiments, we mainly measured the Zeta potential values of
PDA@Fe3O4 and PEI@PDA@Fe3O4 NPs in aqueous suspensions with
the pH range of 2.0–9.0. Fig. S3 shows that the potential values of
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Food Chemistry 293 (2019) 340–347
PDA@Fe3O4 NPs vary with the pH of the solution, and the isoelectric
point is about 4.0. While for PEI@PDA@Fe3O4, the isoelectric point
changes to ∼8.0 although the changing tendency of potential value
versus pH is quite similar. This is mainly because PEI contains a large
amount of imino and amino groups, and so the surface of PEI@PDA@
Fe3O4 would be positively charged with the protonation of these
functional groups in lower pH values.
The XRD spectra are shown in Fig. S4. By comparing the diffraction
pattern to the database of known patterns, it can be concluded that
there are nearly no changes in the number and the position of diffrac-
tion peaks after being coated with PDA and PEI, indicating the coating
layers are amorphous material, and the crystalline magnetic Fe3O4 NPs
do not change during the coating process.
From the hysteresis loops shown in Fig. 2, no obvious hysteresis and
coercivity (or remanence) was observed, suggesting the prepared
magnetic materials were all superparamagnetic and can be easily op-
erated under an external magnetic field. The saturation magnetization
of bare Fe3O4 NPs is 83.2 emu g−1. After being coated by PDA and PEI,
the saturation magnetization decreases to 46.4 emu g−1. However,
these magnetic materials still possessed high magnetic responses and
could be readily separated from solution within 20 s with a magnet.
3.2. Optimization of MSPE procedure
Several important parameters that affect the performance of MSPE
procedure, including solution pH, the amounts of adsorbent, extraction
time, and the type of eluting solvent, were investigated in detail. All the
experiments were performed in triplicate.
3.2.1. Effect of solution pH
The surface charge of PEI@PDA@Fe3O4 NPs and the ionization
degree of the colorants were all strongly affected by the solution pH. In
the investigated pH range, the tested synthetic colorants were nega-
tively charged in most cases, for the presence of sulfonic acid groups in
their molecular structures. Fig. S5 shows all the recoveries reached the
maximum at pH 2. From pH 2–5, the recoveries of tested colorants were
gradually reduced. The tendencies are in accordance with the previous
results from Zeta potential measurement (Fig. S3). At lower pH values,
the surface of the PEI@PDA@Fe3O4 NPs would become more positively
charged, which helps to induce the electrostatic attraction between
adsorbents and the negatively charged colorants. The pH 2.0 was so
eventually selected for further optimization experiments.
3.2.2. Effect of the amount of adsorbents
In order to optimize the amount of PEI@PDA@Fe3O4 NPs, PEI@
PDA@Fe3O4 NPs with the concentration range of 0.1–0.5 mg mL−1
were utilized for the MSPE process. Fig. S6 shows that when the con-
centration was 0.3 mg mL−1, the recoveries of the four colorants
reached maximum. Further increasing the concentration of adsorbents
did not help to improve the extraction efficiency, even worse, the re-
coveries of analytes gradually decreased, probably due to incomplete
desorption. Therefore, 0.3 mg mL−1 was regarded as the optimized
adsorbent concentration and used in further studies.
3.2.3. Effect of the extraction time
To improve the precision and sensitivity of MSPE, it is necessary to
provide enough contact time to ensure the adsorption equilibrium. So,
the impact of the extraction time on extraction efficiency was per-
formed by varying the shaking time from 10 to 50 min. It is found that
the recoveries of all analytes increased with the increasing shaking time
up to 20 min, further prolonging contact time showed no obvious im-
provements (see Fig. S7). For saving time, 20 min was eventually se-
lected as the optimized extraction time in further studies.
3.2.4. Effect of the type of eluting solvent
In order to obtain higher recoveries of target analytes, different
H. Chen, et al.
Fig. 1. TEM images of PDA@Fe3O4 MNPs: (a, b) and PEI@PDA@Fe3O4 MNPs: (c, d).
Fig. 2. Hysteresis loops of Fe3O4 NPs (a) and PEI@PDA@Fe3O4 NPs (b).
types of eluents were investigated, mainly including pure CH3OH, PBS
(10 mM, pH 10), CH3OH-10 mM NH4Cl (pH 10, v/v = 1:1), CH3OH-
NH3·H2O (70:30, v/v), CH3CN-NH3·H2O with different ratios. As re-
presented in Fig. S8, lowest recoveries were obtained when using al-
kaline phosphate buffer as desorption solvent, while CH3CN-NH3·H2O
mixtures showed higher recovery. For better results, CH3CN-NH3·H2O
mixtures with different ratios as eluting solutions were further eval-
uated. As demonstrated in Fig. S8, CH3CN-NH3·H2O (70:30, v/v) pro-
vided the highest recoveries for all the colorants. Therefore, CH3CN-
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Food Chemistry 293 (2019) 340–347
NH3·H2O (70:30, v/v) was eventually regarded as the optimized eluting
solution for the subsequent studies.
3.3. Adsorption mechanism studies
3.3.1. Adsorption kinetics
In order to study the variation of adsorption amount with adsorp-
tion time, the adsorption kinetics experiments were carried out at room
temperature (25 °C). As shown in Fig. S9, the adsorption amounts in-
creased rapidly during the initial period of time, and then slowly
reached stable values in less than 30 min, indicating the adsorption
occurred mainly on the surface of the adsorbent. The kinetic data are
subjected to pseudo-first-order and pseudo-second-order equations and
they are usually expressed as follows (Fu et al., 2015):
Pseudo - first - order: ln(qe − qt ) = ln qe − k1 t
t 1 1
Pseudo - second - order: = + t
qt k2 qe qe
where qe (mg g−1) is the equilibrium adsorption amount, qt (mg g−1) is
the amount of adsorption at time t (min), k1 (min−1) and k2
(g mg−1 min−1) are rate constants of pseudo-first-order and pseudo-
second-order equations, calculated from the nonlinear curve fitting of
the plot of qt versus t respectively.
Fit curves and relevant parameters are shown in Fig. S10 and Table
S1 respectively. It can be observed that the fit curves of pseudo-second-
order for all the colorants are better than those of pseudo-first-order,
with the regression coefficients (R2) all higher than 0.99. Therefore, the
adsorption processes of colorants on PEI@PDA@Fe3O4 NPs can be
better illustrated by pseudo-second-order model, showing the chemical
H. Chen, et al. Food Chemistry 293 (2019) 340–347

Table 1
The contents of four colorants in actual samples.
Samples Ingredients Colorants Contents Added Recovery (%, n = 3)
detected (g kg−1) (g mL−1)

Candy (grape) White sugar, glucose syrup, oxidized vegetable oil, grape juice (2.5%), Amaranth 0.0163 0.01 103.32 ± 2.32
cream, food additives (lactic acid, sodium lactate, soy lecithin, titanium 0.10 101.48 ± 1.54
dioxide, amaranth, bright blue), maltodextrin, food flavor, edible salt 0.30 98.56 ± 1.01

Candy (strawberry) White sugar, glucose syrup, oxidized vegetable oil, strawberry juice (2.5%), Allure Red 0.0246 0.01 110.50 ± 4.68
cream, food additives (lactic acid, sodium lactate, soybean phospholipid, 0.10 108.73 ± 3.10
titanium dioxide, Allure red), maltodextrin, food flavor, edible salt 0.30 101.54 ± 2.67

Jelly (strawberry) Water, apple juice (≥15%), white sugar, fructose syrup, konjac flour, food Ponceau 4R 0.0031 0.01 109.75 ± 5.61
additives (calcium lactate, carrageenan, potassium chloride, citric acid, 0.10 99.67 ± 3.45
malic acid, sodium citrate, sodium cyclamate, potassium sorbate, Ponceau 0.30 93.27 ± 1.78
4R), food flavor

Carbonated drinks Water, fructose syrup, white sugar, maltodextrin, food additives (carbon Sunset yellow 0.0118 0.01 98.45 ± 6.48
(orange) dioxide, citric acid, sodium hexametaphosphate, potassium sorbate, food 0.10 97.64 ± 5.20
flavor, acesulfame, vitamin C, sucralose, Sunset yellow, Lemon yellow, 0.30 93.69 ± 2.43
Lemon Sodium, Ponceau 4R) Ponceau 4R 0.0025 0.01 103.78 ± 1.64
0.10 97.51 ± 3.56
0.30 94.50 ± 1.20

calculated by the following equation (Bai et al., 2015):

Fig. 3. The chromatograms of (a) strawberry flavored jelly sample, (b) straw-
berry flavored jelly sample after MSPE and (c) the sample spiked with four
colorants at 0.1 mg L−1 after MSPE. Peaks: (1) Amaranth (2) Ponceau 4R (3)
Sunset yellow (4) Allure red. Detection wavelength: 515 nm.
adsorption is the rate-limiting step in the adsorption process (Zhang
et al., 2016).
3.3.2. Adsorption isotherm
In order to better understand the adsorption mechanism of the PEI@
PDA@Fe3O4 NPs toward colorants investigated, the adsorption iso-
therms for Amaranth, Ponceau 4R, Sunset yellow and Allure red with
various initial concentrations were carried out at room temperature. As
shown in Fig. S11, at the first stage, the adsorption capacities of four
colorants on PEI@PDA@Fe3O4 NPs increased significantly, and then
increase slightly with the increase of initial colorant concentration, and
finally reached plateaus. These data can be analyzed by Langmuir iso-
thermal and Freundlich isothermal models, which are usually expressed
as follows (Fu et al., 2015),
ce c 1
Langmuir equation: = e +
qe qm qm KL
1
Freundlich equation: ln qe = ln F + ln Ce
n
RL reflects the basic characteristic of Langmuir model, and can be
1
RL =
1 + KL C0
where Co (mg L−1) is the initial concentration of colorant in solution, ce
(mg L−1) is the concentration of the colorant in solution after reaching
equilibrium. While qe (mg g−1) represents the amount of colorant ab-
sorbed on the adsorbent at equilibrium, qm (mg g−1) denotes the
maximum adsorption capacity corresponding to complete monolayer
coverage. KL is the adsorption equilibrium constant; KF and n are the
Freundlich constants.
The corresponding adsorption parameters can be acquired from the
fitting curves of Langmuir model (Fig. S12a) and Freundlich model (Fig.
S12b). By comparing the regression coefficients (R2) values (shown in
Table S2), it can be concluded that Langmuir model fitted better to
experimental data than Freundlich model. Therefore, the adsorption
process could be better explained by the Langmuir model, indicating
that the monolayer adsorption occurred at homogeneous surfaces of
PEI@PDA@Fe3O4 NPs (Zhang et al., 2016). According to the Langmuir
data, the maximum adsorption capacities for four colorants, Amaranth,
Ponceau 4R, Sunset yellow and Allure Red were 93.54, 50.28, 17.83
and 41.09 mg g−1 respectively.
When the value of RL is equal to 0, higher than 0 but lower than 1,
equal or higher than 1, the adsorption process is irreversible, favorable
and linear respectively (Li et al., 2016). Moreover, the adsorption
process is more favorable when the RL value is smaller (0 < RL < 1).
As shown in Table S2, the values of RL for four colorants are all smaller
than 1, indicating that the adsorption process of four colorants on PEI@
PDA@Fe3O4 NPs are favorable under the current experimental condi-
tions.
3.4. Validation of the MSPE-HPLC method
Based on the linear range, limit of detection (LOD) and limit of
quantitation (LOQ), the analytical properties of the established MSPE-
HPLC method were examined. Firstly, a four-colorant mixed standard
solution (1000 μg L−1 for each component) was prepared. The mixed
standard was then diluted to working solutions (0.2, 0.4, 0.6, 0.8, 1, 5,
10, 20, 40, 80, 160, 320, 400, 450, 500, 550, 600 μg L−1) with dilute
hydrochloric acid. Wide linearity ranges (chromatographic peak area
versus concentration) were obtained for all the colorants with
0.60–500 μg L−1. For all the colorants, the measured LOD (S/N = 3:1)
and LOQ (S/N = 10:1) were 0.15–0.20 μg L−1 and 0.50–0.70 μg L−1,
respectively. The precisions were measured and the obtained RSDs
(n = 3) were in the range of 2.56–5.67 (%), and the regression

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 H. Chen, et al.

Table 2
Comparison of the proposed method with other reported methods for the detection of synthetic colorants.
Analytical method Adsorbent/extractant Colorants Samples Extraction time Linearity (μg L−1) LOD (μg L−1) References
(min)

M-dSPE-UFLC-MS/MS NH2-LDC-MP Amaranth, Allure Red Wines and drinks – 5–1000 0.45–15 Chen et al. (2014)
MSPE-HPLC Fe3O4@SiO2@IL NPs Rhodamine B Chili powder, Chinese prickly ash 10 0.50–150 0.08 Chen and Zhu (2016)
UV–vis Spectrophotometry Fe3O4/sodium dodecyl sulfate Methylene blue Shrimp, Tap water Karaj river – 8–2212 2.5 Fasih Ramandi and
Shemirani (2015)
MSPE-fluorescence Fe3O4-poly(aniline-naphth Rhodamine B Dish washing foam, Dish washing liquid 10 0.35–5 0.1 Bagheri, Daliri, and
lamine)-based nanocomposite shampoo, Pencil lead Roostaie (2013)
HPLC-DAD-ESI-MS – Amaranth, Ponceau 4R, Sunset yellow Drink and ginger 15 400–40,000 0.3–0.4 Ma, Luo, Chen, Su, and
Yao (2006)
SPE-CE Polyamide Carminic acid, Tartrazine, Fast green Milk 3.39–7.91 1000–500000 50–400 Huang et al. (2002)
FCF, Brilliant blue FCF, etc.

345
HPLC-DAD – Tartrazine, Quinoline Yellow, Sunset Fruit flavored drink, Fruit flavored liqueur, – – 1.59–22.1 Minioti et al. (2007)
yellow, Carmoisine, Amaranth, Icing sugar, Sweets, Blackberry flavored jam
Ponceau 4R, etc.
HPLC-DAD Polyamide Lemon yellow, New Red, Amaranth, Juice drinks, Alcohol, Hard candy, Candied – – 0.2–0.5 (mg (GB5009.35-2016)
Ponceau 4R, Sunset yellow, Temptation fruit, Starch jelly, Chocolate beans and Colored kg−1)
red, etc. sugar-coated products
MSPE-HPLC-DAD PEI@PDA@Fe3O4 NPs Amaranth, Ponceau 4R Candy, Jelly and Carbonated drink 20 0.60–500 0.15–0.20 This work
Sunset yellow, Allure red

M-dSPE: magnetic dispersive solid-phase extraction.

UFLC–MS/MS: ultra-fast liquid chromatography-tandem quadrupole mass spectrometry.
NH2-LDC-MP: An amino-functionalized low degrees of cross-linking magnetic polymer.
IL: ionic liquid.
ESI-MS: electrospray ionization-mass spectrometry.
Food Chemistry 293 (2019) 340–347
H. Chen, et al.
coefficients (R2) of four colorants were all higher than 0.999. All the
data indicate the established method is feasible for the analysis of trace
amounts of colorants.
The calculation formula for enrichment factor (EF) is as follows:
Cen V C V
EF = = ⎛ 0 ⎞ × ER, ER = ⎛ en en ⎞ × 100%
⎜ ⎟ ⎜ ⎟
C0 ⎝ Ven ⎠ ⎝ C0 V0 ⎠
where C0, V0 and Cen, Ven are the concentration and volume of analyte
before and after MSPE process, and ER is the extraction recovery. By
calculation, the EF values for four colorants are all above 184, which
proves that the enrichment method is highly efficient.
3.5. Reusability of PEI@PDA@Fe3O4 NPs
In this study, the same batch of PEI@PDA@Fe3O4 NPs were sub-
jected to cyclic utilization for the evaluation of the reusability and
stability of the synthesized materials. In this procedure, PEI@PDA@
Fe3O4 NPs were washed with dilute hydrochloric acid (pH 2) for 3 times
prior to next use (monitored by UV–Vis spectrometer). It was found that
the recoveries decreased only ∼10% even after six cycles, suggesting
that the synthesized PEI@PDA@Fe3O4 NPs were relatively stable, and
can be regarded as promising adsorbents for the effective extraction of
colorants (Fig. S13).
3.6. Real sample analysis
In order to evaluate the applicability of the proposed method for the
analysis of actual samples, two types of candies, jelly and carbonated
drink from local supermarkets were analyzed, and the results are
showed in Table 1. The representative chromatograms of strawberry
flavored jelly sample, strawberry flavored jelly sample after MSPE and
the sample spiked with four colorants at 0.1 mg L−1 after MSPE were
presented in Fig. 3. The contents of four synthetic colorants detected in
these foods comply with the provisions of the hygiene standards for
food additives (GB2760), and no excessive levels have been found. To
further verify the accuracy of the proposed method, the recoveries of
the spiked samples at three levels of 0.01 μg mL−1, 0.1 μg mL−1, and
0.3 μg mL−1 were studied. As shown in Table 1, the recovery rates were
in the range of 93.27%–109.75% with RSDs less than 6.5%. All these
data demonstrate the applicability and reliability of the proposed
method for the analysis of colorants in actual samples.
The proposed method based on MSPE (PEI@PDA@Fe3O4 NPs)-
HPLC-DAD was compared to other reported methods (see Table 2).
LODs of the present method are comparable to some sensitive detection
methods, such as MS and fluorescence. Besides, wide linear region was
obtained for the proposed analysis method. The sensitivity of this
method is at least one order of magnitude higher than those HPLC (or
CE) methods using traditional SPE pretreatment technique.
4. Conclusion
In this study, a novel magnetic material (PEI@PDA@Fe3O4 NPs)
was successfully synthesized and used as adsorbent for the pre-con-
centration of several representative synthetic colorants. High con-
centration factor, good recoveries and great reproducibility were ob-
tained for these colorants in food samples. The experimental data of this
study indicate that PEI@PDA@Fe3O4 NPs can be used as a highly ef-
ficient MSPE material for the analysis of trace colorants in food sample,
when hyphenating with HPLC. In comparison with those chromato-
graphic methods for the analysis of colorants reported before, the es-
tablished MSPE-HPLC method showed some obvious advantages, such
as simple manipulation, wide linearity ranges and high sensitivity.
346
Food Chemistry 293 (2019) 340–347
Acknowledgments
Financial support from the National Natural Science Foundation of
China (21005054 and 201506147) and State Key Laboratory of
Medicinal Chemical Biology (Nankai University, 201603004) is grate-
fully acknowledged.
Conflict of interest
The authors declare no potential conflict of interest.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.foodchem.2019.04.111.
References
Alves, S. P., Brum, D. M., Ecbde, A., & Pereira Netto, A. D. (2008). Determination of
synthetic dyes in selected foodstuffs by high performance liquid chromatography
with UV-DAD detection. Food Chemistry, 107(1), 489–496.
Bagheri, H., Daliri, R., & Roostaie, A. (2013). A novel magnetic poly(aniline-naphthyla-
mine)-based nanocomposite for micro solid phase extraction of rhodamine B.
Analytica Chimica Acta, 794(17), 38–46.
Bai, L., Li, Z., Zhang, Y., Wang, T., Lu, R., Zhou, W., ... Zhang, S. (2015). Synthesis of
water-dispersible graphene-modified magnetic polypyrrole nanocomposite and its
ability to efficiently adsorb methylene blue from aqueous solution. Chemical
Engineering Journal, 279, 757–766.
Chai, W., Wang, H., Zhang, Y., & Ding, G. (2016). Preparation of polydopamine-coated
magnetic nanoparticles for dispersive solid-phase extraction of water-soluble syn-
thetic colorants in beverage samples with HPLC analysis. Talanta, 149, 13–20.
Chen, J., & Zhu, X. (2016). Magnetic solid phase extraction using ionic liquid-coated
core–shell magnetic nanoparticles followed by high-performance liquid chromato-
graphy for determination of Rhodamine B in food samples. Food Chemistry, 200,
10–15.
Chen, Q. C., Mou, S. F., Hou, X. P., Riviello, J. M., & Ni, Z. M. (1998). Determination of
eight synthetic food colorants in drinks by high-performance ion chromatography.
Journal of Chromatography A, 827(1), 73.
Chen, X. H., Zhao, Y. G., Shen, H. Y., Zhou, L. X., Pan, S. D., & Jin, M. C. (2014). Fast
determination of seven synthetic pigments from wine and soft drinks using magnetic
dispersive solid-phase extraction followed by liquid chromatography-tandem mass
spectrometry. Journal of Chromatography A, 1346, 123–128.
Cheng, C., Li, S., Zhao, J., Li, X., Liu, Z., Ma, L., ... Zhao, C. (2013). Biomimetic assembly
of polydopamine-layer on graphene: Mechanisms, versatile 2D and 3D architectures
and pollutant disposal. Chemical Engineering Journal, 228(28), 468–481.
Deng, H., Li, X., Peng, Q., Wang, X., Chen, J., & Li, Y. (2005). Monodisperse magnetic
single-crystal ferrite microspheres. Angewandte Chemie, 44(18), 2782.
Ertaş, E., Özer, H., & Alasalvar, C. (2007). A rapid HPLC method for determination of
Sudan dyes and Para Red in red chilli pepper. Food Chemistry, 105(2), 756–760.
Fasih Ramandi, N., & Shemirani, F. (2015). Surfacted ferrofluid based dispersive solid
phase extraction; a novel approach to preconcentration of cationic dye in shrimp and
water samples. Food Chemistry, 185, 398–404.
Feng, L., Cao, M., Ma, X., Zhu, Y., & Hu, C. (2012). Superparamagnetic high-surface-area
Fe3O4 nanoparticles as adsorbents for arsenic removal. Journal of Hazardous
Materials, 217–218, 439–446.
Fu, J., Chen, Z., Wang, M., Liu, S., Zhang, J., Zhang, J., ... Xu, Q. (2015). Adsorption of
methylene blue by a high-efficiency adsorbent (polydopamine microspheres):
Kinetics, isotherm, thermodynamics and mechanism analysis. Chemical Engineering
Journal, 259, 53–61.
Gao, J. F., Chen, R. N., Su, K., Zhang, Q., & Peng, Y. Z. (2009). Inactive aerobic granules as
biosorbent for the removal of anionic azo dye Acid Red 14 from aqueous solution:
Isotherms, kinetics and thermodynamics. Fresenius Environmental Bulletin, 18(9),
1681–1691.
Gosetti, F., Gianotti, V., Angioi, S., Polati, S., Marengo, E., & Gennaro, M. C. (2004).
Oxidative degradation of food dye E133 brilliant blue FCF liquid chromatography-
electrospray mass spectrometry identification of the degradation pathway. Journal of
Chromatography A, 1054(1–2), 379–387.
Huang, H. Y., Shih, Y. C., & Chen, Y. C. (2002). Determining eight colorants in milk
beverages by capillary electrophoresis. Journal of Chromatography A, 959(1–2),
317–325.
Jun-Kai, G., Li-An, H., Guang-Hui, Z., & Ping, G. (2015). Facile functionalized of SBA-15
via a biomimetic coating and its application in efficient removal of uranium ions from
aqueous solution. Journal of Hazardous Materials, 286, 325–333.
Kiseleva, M. G., Pimenova, V. V., & Eller, K. I. (2003). Optimization of conditions for the
HPLC determination of synthetic dyes in food. Journal of Analytical Chemistry, 58(7),
685–690.
Knill, C. J., & Kennedy, J. F. (1996). Food additive toxicology: Edited by J.A. Magu and
A.T. Tu, Marcel Dekker, Inc., New York, 1995. viii + 542pp. Price $175.00. ISBN 0-
8427-9245-9. Carbohydrate Polymers, (4), 294–295.
Li, K., Li, P., Cai, J., Xiao, S., Yang, H., & Li, A. (2016). Efficient adsorption of both methyl
H. Chen, et al.
orange and chromium from their aqueous mixtures using a quaternary ammonium
salt modified chitosan magnetic composite adsorbent. Chemosphere, 154, 310–318.
Long, Y., Xiao, L., & Cao, Q. (2017). Co-polymerization of catechol and polyethylenimine
on magnetic nanoparticles for efficient selective removal of anionic dyes from water.
Powder Technology, 310, 24–34.
Ma, M., Luo, X., Chen, B., Su, S., & Yao, S. (2006). Simultaneous determination of water-
soluble and fat-soluble synthetic colorants in foodstuff by high-performance liquid
chromatography–diode array detection–electrospray mass spectrometry. Journal of
Chromatography A, 1103(1), 170–176.
Minioti, K. S., Sakellariou, C. F., & Thomaidis, N. S. (2007). Determination of 13 synthetic
food colorants in water-soluble foods by reversed-phase high-performance liquid
chromatography coupled with diode-array detector. Analytica Chimica Acta, 583(1),
103–110.
Nayab, S., Farrukh, A., Oluz, Z., Tuncel, E., Tariq, S. R., Ur, R. H., ... Yameen, B. (2014).
Design and fabrication of branched polyamine functionalized mesoporous silica: An
efficient absorbent for water remediation. ACS Applied Materials & Interfaces, 6(6),
4408–4417.
Rodríguez, A. R. C., Saizposeu, J., Garcíapardo, J., García, B., Lorenzo, J., Ojeajiménez, I.,
... Sánchez, A. (2016). Biocompatible polydopamine-like particles for the removal of
347
Food Chemistry 293 (2019) 340–347
heavy metals at extremely low concentrations. RSC Advances, 6(46), 40058–40066.
Soponar, F., Moţ, A. C., & Sârbu, C. (2008). Quantitative determination of some food dyes
using digital processing of images obtained by thin-layer chromatography. Journal of
Chromatography A, 1188(2), 295–300.
Wang, Y., Wang, S., Niu, H., Ma, Y., Tao, Z., Cai, Y., & Meng, Z. (2013). Preparation of
polydopamine coated Fe3O4 nanoparticles and their application for enrichment of
polycyclic aromatic hydrocarbons from environmental water samples. Journal of
Chromatography A, 1283(3), 20–26.
Zafar, I., Samar, A., Musharraf, M., & Lai, E. P. C. (2011). Rapid CE-UV binding tests of
environmentally hazardous compounds with polymer-modified magnetic nano-
particles. Electrophoresis, 32(16), 2181–2187.
Zhang, Y. Z., Li, J., Zhao, J., Bian, W., Li, Y., & Wang, X. J. (2016). Adsorption behavior of
modified Iron stick yam skin with polyethyleneimine as a potential biosorbent for the
removal of anionic dyes in single and ternary systems at low temperature. Bioresource
Technology, 222, 285–293.
Zheng, T., Yang, Z., Gui, D., Liu, Z., Wang, X., Dai, X., ... Chen, L. (2017). Overcoming the
crystallization and designability issues in the ultrastable zirconium phosphonate
framework system. Nature Communications, 8, 15369.