Food Control 150 (2023) 109775

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Blue/red dual emission based ratiometric fluorescent intelligent labels for

real-time food freshness monitoring
Shuo Yang b, Jiang Lou a, b, *, Limin Jing b, Qijun Ding b, Xia Li b, Yifei Jiang b, Zhuqing Liu b,
Wenjia Han a, b, **
a
Key Laboratory of Pulp and Paper Science & Technology of Ministry of Education, Qilu University of Technology, Shandong Academy of Sciences, Jinan, 250353, PR
China
b
State Key Laboratory of Biobased Material and Green Papermaking, Qilu University of Technology, Shandong Academy of Sciences, Jinan, 250353, PR China

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

Keywords: Fluorescent sensor technology is used in food safety to monitor seafood spoilage with the naked eye in real time
Ratiometric fluorescent aerogels due to its ease of operation, low cost and high degree of visualisation. Therefore, in this work, carbon quantum
Collagen fiber dots (CDs) as pH indicators and protoporphyrin IX (PpIX) as an internal reference were covalently grafted onto
Carbon quantum dots
collagen fibres (CF) to design collagen fibre-based ratiometric fluorescent aerogels (CF-CP) with excellent amine
Food freshness monitoring
response. As the alkaline environment causes a molecular conformational change in CDs and a decrease in
fluorescence intensity, PpIX is unaffected, increasing its CF-CP colour response from blue to deep red with
increasing ammonia concentration, with detection limits as low as 1.0 × 10− 6 mg/L. CF-CP responds well to a
wide range of biogenic amines (BAs), with the best selectivity for tyramine. In addition, the aerogel is
biocompatible, degradable and colour responsive. The development of this smart fluorescent aerogel label
provides a new platform for food quality monitoring and has wide application prospects in the field of food
safety.

1. Introduction fluorescence sensing technology has been developed to enable real-time

Food safety is a long-term global concern, and food spoilage is one of
the most important issues for consumers and the food industry. How­
ever, food spoilage cannot be immediately identified by sight, smell or
taste and can only be judged by shelf life, resulting in a large amount of
food being wasted (Hu, Ma, Zhang, Che, & Zhao, 2016; Leng, Zhao, Yin,
& Ye, 2015). Biogenic amines(BAs), produced by microbial fermenta­
tion, are recognised as an effective and convenient indicator for
detecting food spoilage (Zhong et al., 2018). In this respect, numorous
methods have been developed to monitor BAs. Among these, electro­
chemical biosensors (Xu, Guo, Yang, Gao, & Song, 2021; Y. H. Zhang
et al., 2021), capillary electrophoresis, and gas-phase liquid chroma­
tography have provided accurate data reference values for food safety
(Sentellas, Nunez, & Saurina, 2016). However, their use for real-time
monitoring applications is severely hampered by shortcomings such as
being time-consuming, expensive and complex to perform. As a result,
monitoring of food spoilage by the naked eye, solving the disadvantages
of traditional detection methods such as high cost and complexity of
operation (O. Bunkoed, Raksawong, Chaowana, & Nurerk, 2020; Tang
et al., 2021).
Among them, based on the principles of structural changes, proton­
ation and deprotonation of fluorescent colour-emitting groups and en­
ergy leaps leading to the decay or enhancement of fluorescence, they are
widely used for the detection and adsorption of harmful substances such
as heavy metal ions, organic pesticides and organic volatile substances
(Cai et al., 2019; Jing et al., 2022; Quan et al., 2021). However, these
optical sensors are based on changes in fluorescence intensity of only
one luminous cluster and often fail to achieve high sensitivity and
outstanding colour rendering (Opas Bunkoed, Donkhampa, & Nurerk,
2020; Chen et al., 2022; Pongprom, Chansud, & Bunkoed, 2022). This is
not only attributable to the fact that the luminophores are affected by
the external environment, but also that in most cases the fluorescence

* Corresponding author. Key Laboratory of Pulp and Paper Science & Technology of Ministry of Education, Qilu University of Technology, Shandong Academy of
Sciences, Jinan, 250353, PR China.
** Corresponding author. Key Laboratory of Pulp and Paper Science & Technology of Ministry of Education, Qilu University of Technology, Shandong Academy of
Sciences, Jinan, 250353, PR China.
E-mail addresses: jlou@qlu.edu.cn (J. Lou), hwj200506@163.com (W. Han).

https://doi.org/10.1016/j.foodcont.2023.109775
Received 17 December 2022; Received in revised form 18 March 2023; Accepted 30 March 2023
Available online 30 March 2023
0956-7135/© 2023 Elsevier Ltd. All rights reserved.
S. Yang et al.
Fig. 1. Proportional fluorescent aerogel (CF-CP) preparation process.
intensity of the luminophores varies very little and that the human eye
has limited sensitivity to changes in fluorescence brightness, which may
increase experimental error or even lead to failure of the naked eye
detection mode (X. J. Liu, Zhang, Bing, & Shangguan, 2014). Propor­
tional fluorescence overcomes the disadvantages of low sensitivity and
poor colour development of a single fluorescent colour-emitting group,
making it a more promising candidate. More importantly, ratiometric
fluorescence overcomes the interference of the external environment
and signal-to-noise ratio by presenting a distinct fluorescence colour
change or even the naked eye to identify the analyte by background
reference (Quan et al., 2021). However, its difficult synthesis, poor
processing properties, high cost and poor reactivity still limit its prac­
tical application.
Bio-based materials are of great interest due to their green, sustain­
able, renewable, biodegradable and processable nature. Of these,
Collagen fibres are derived from animal meat tissue and are large mol­
ecules with a spiral structure formed by the polymerisation of collagen
into collagenous fibrils. Its surface contains a large number of functional
groups such as –NH2, –COOH, –OH and –CO–NH-, which provide
favorable binding sites for small molecules without surface modification
(Wells et al., 2013). The excellent luminescence of fluorescent materials
is due to the covalently attached fluorescent groups at the active sites of
biobased materials, which avoids the phenomenon of fluorescence
group aggregation induced quenching (ACQ) (Tian et al., 2016). In
addition, the composite not only has excellent luminescence, structural
and performance stability but also avoids secondary environmental
contamination due to the physical shedding of the fluorescent material.
Currently, there are more bio-based fluorescent films for the detection of
BAs, but the dense structure and low porosity of the film material results
in low sensitivity for gas detection, poor colour rendering and inability
to monitor food spoilage in real time. Aerogels facilitate the efficient
capture of harmful substances due to their high porosity and high spe­
cific surface area. However, fluorescent aerogels based on collagen fi­
bres have rarely been reported in food detection.
In this study, a collagen fibre-based proportional fluorescent aerogel
(CF-CP) was designed and used to assess the freshness of shrimps in a
visual way. Carbon quantum dots (CDs, blue, indicator) and protopor­
phyrin IX (PpIX, red, background reference) were covalently bound to
collagen fibrils (CF), respectively. The initial fluorescence colour was
adjusted according to different mass ratios of the collagen fibre de­
rivatives CF-CD and CF-PpIX mixed to select the best colour ratio aerogel
for the experimental study. The change in colour of the proportional
fluorescent aerogel from blue to deep red caused by shrimp spoilage
alkaline gas, and thus the edibility of the food according to the degree of
colour change.
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Food Control 150 (2023) 109775
2. Experimental section
2.1. Materials
Collagen fiber was obtained from Jinan Shengquan Group (Jinan,
China). Dialysis bag (Mw = 10000), Polyethyleneimine (PEI, Mw =
25000), N-(3-dimethylaminopropyl)-N -ethylcarboxydiimine hydro­
chloride (EDC, 98%), N-hydroxysuccinimide (NHS, 98%), protopor­
phyrin IX (PpIX, >95%), N, N -carbonyldiimidazole (CDI, AR, 99%),
trimethylamine (AR, 25%), morpholine (AR, 99.5%), 1,4-butanedi­
amine (AR, 98%), diethylamine (>99.5%, GC), tyramine (AR, 98%),
benzylamine (AR, 99%), triethylamine (>99.5%, GC), aminopentane
(AR, 98%), ammonia (AR, 25% a), dimethylamine (AR, 40%), pyrroli­
dine (AR, 98%), dimethyl sulfoxide (DMSO, AR, >99.9%) were pur­
chased from Shanghai Maclean Biochemical Technology Co. and used
directly without any treatment.
2.2. Preparation of carbon quantum dots (CDs)
Dissolve 0.2 g of citric acid and 1.42 g of polyethyleneimine in 120
mL of deionised water with stirring. The mixture was placed in a 200mL
PTFE stainless steel autoclave and reacted at 180 ◦ C for 24 hours. At the
end of the reaction, the product was cooled to room temperature and
poured into a dialysis bag (Mw = 10000) for purification, changing the
deionised water every 6 hours for a total of 72 hours to avoid interfer­
ence from unreacted molecules in the solution.
2.3. Preparation of collagen fibers - carbon quantum dots (CF-CD)
The 3.5g collagen fibers (after freeze-drying) into a 500ml beaker of
350 mL deionised water and stir at 60 ◦ C until a uniform suspension. 10
mL CDs was added to the collagen fiber suspension and mixed well. 24
mg of NHS and 30 mg of EDC catalyst were added to react at 40 ◦ C for 12
hours. After the reaction was completed, the mixture was cooled to a gel
state. The supernatant was washed with absolute ethanol to a non-
fluorescent state. The final samples were vacuum dried at 50 ◦ C for 24
hours and sealed at 4 ◦ C.
2.4. Preparation of collagen fiber-protoporphyrin IX (CF-PpIX)
The 3.5g collagen fibers were dissolved in 350 mLDMSO and stirred
to a uniform suspension at 45 ◦ C. 0.1g of PpIX was first activated with
CDI at 45 ◦ C for 4 hours, then mixed with the collagen fiber suspension
and reacted at 80 ◦ C for 24 hours. At the end of the reaction the pre­
cipitate was cooled to room temperature. Then precipitation and
S. Yang et al.
Fig. 2. (a) UV, excitation and emission spectra of CDs. Inset: Fluorescent images of CDs under daylight and UV light. (b) Fluorescence intensity of CDs at pH = 7–14.
(c) Emission spectra of CDs. (d) UV, excitation and emission spectra of PpIX. Inset: Fluorescent images of PpIX under daylight and UV light. (e) Fluorescence intensity
of PpIX at pH = 7–14. (f) Emission spectra of PpIX at excitation wavelengths from 250 nm to 400 nm.
washing with absolute ethanol until the supernatant has no fluores­
cence, and then vacuum-dried at 50 ◦ C for 24 hours and stored at 4 ◦ C.
2.5. Preparation of ratiometric fluorescent aerogels
CF-CD and CF-PpIX were dissolved at a mass ratio of 39:1 in deion­
ised water at 40 ◦ C to obtain a 1 wt% suspension. The mixed solution
was then cooled to room temperature and placed in a 3cm × 3cm mold
for freeze-drying to obtain ratiometric fluorescent aerogels as shown in
Fig. 1.
2.6. Preparation and testing of solutions with different ammonia
concentrations
The 1 wt% suspension of CF-CD, CF-CD mixed with CF-PpIX were
prepared separately in deionised water. Industrial ammonia was diluted
with different amounts of deionised water to different concentrations
and different types of biogenic amines (tyramine, benzylamine, mor­
pholine, aminopentane, trimethylamine, diethylamine, ammonia, trie­
thylamine, pyrrolidine, dimethylamine) to 1.0 × 10− 2 mg/L solution.
Finally, CF-CD, CF-CD and CF-PpIX mixed solutions were mixed with
different concentrations and types of ammonia solutions at a volume
ratio of 1:2, and their fluorescence intensities were subsequently
recorded on a fluorescence spectrometer (F-7100, Hitachi, Japan)(Quan
et al., 2021).
2.7. Application testing for freshness of seafood
The fluorescent aerogels (CF-CP) were placed in petri dishes with
shrimps and sealed with cling film. the colour change of CF-CP aerogels
in three different temperature environments freezer (− 18 ◦ C), fresh
room (4 ◦ C) and room temperature (25 ◦ C) was used to determine the
degree of spoilage of the food, and three parallel samples were made for
each group. The colour changes of the fluorescent aerogels at different
temperatures were recorded using a One Plus Nine smartphone and RGB
and Lab values were recorded using a smartphone (colour identifier) for
multi-point analysis.
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Food Control 150 (2023) 109775
2.8. Characterization
The UV absorption of CDs and PpIX in the range 200–600 nm was
recorded by UV spectrophotometer (UV-2700, Shimadzu, Japan). The
fluorescence intensity, excitation and emission spectra of the two fluo­
rophores under different conditions were measured using a fluorescence
spectrophotometer (F-7100, Hitachi, Japan). The stability of the aero­
gels was analysed using a thermogravimetric analyser (TG-50, TA In­
struments, USA). The crystalline structure of the grafted aerogels was
analysed by an X-ray diffraction analyser (XRD) (Smartlab SE, Rigaku,
Japan). The structural morphology of the fluorescent aerogels was
studied using scanning electron microscopy (SEM) (JSM-6700F, JEOL,
Japan), laser confocal microscopy (CLSM) (LSM800, Zeiss, Germany)
and micro-CT (vivaCT40, SCANCO Medical AG, Switzerland). X-ray
photoelectron spectroscopy (XPS) (Thermo Fisher, ESCALAB 250XI+,
USA) was used to determine the chemical bonds and their bonding en­
ergy in the fluorescent aerogels chemical bonds and their bond energies
were determined.
3. Results and discussion
3.1. Fluorescence properties of CDs and IX (PpIX)
The fluorescence behaviour of two different fluorophores under
different conditions was investigated using UV–Vis and fluorescence
spectroscopy. As shown in Fig. 2a, under daylight irradiation the CDs
appeared as a light yellow solution and under UV = 365 nm the CDs
showed blue fluorescence. Under excitation at Ex = 400 nm, a visible
blue emission peak at Em = 475 nm was obtained for the CDs solution.
This is due to the appearance of two characteristic peaks in the UV at
243 nm and 360 nm, resulting from π-π* of C– – C and C–– N and n-π* of
C–– O in the sp2 structural region of the aromatic ring (Song et al., 2016).
As shown in Fig. S1, the overall trend of fluorescence intensity of CDs
increased with decreasing pH when pH = 1–6. However, when the so­
lution was alkaline at pH = 7–14, the fluorescence intensity of CDs
decreased significantly with increasing alkalinity, and the peak position
at 460 nm remained the same, as shown in Fig. 2b. This demonstrates
the differential response of CDs to pH. In Fig. 2c, the peak position of the
CDs remains constant at different wavelengths of excitation, causing
S. Yang et al. Food Control 150 (2023) 109775

Fig. 3. (a) Structural formulae for the reactions of CF with CDs and PpIX, respectively. (b) Wide-scan XPS spectrum of CF, CF-CD, CF-PpIX. (c) Deconvolution peaks
of the C1s XPS spectrum of CF, (d) CF-CD, (e) CF-PpIX. (f) Deconvolution peaks of the N1s XPS spectrum of CF, (g) CF-CD and (h) CF- PpIX.

only a change in fluorescence intensity. As shown in Fig. 2d, PpIX
appeared dark red in daylight, and under UV = 365 nm irradiation, the
PpIX solution appeared distinctly bright red. At an excitation wave­
length of Ex = 310 nm, the PpIX solution emits a red emission peak at
Em = 635 nm. The UV of PpIX peaks at 407 nm, which was attributed to
the tetrapyrrole macrocyclic conjugated aromatic heterocyclic structure
of PpIX. PpIX does not respond to pH and the position of its emission
peak does not change at different excitation wavelengths, as shown in
Fig. 2e–f. This is compatible with the effect presented by the CDs above.
The may be related to the nano-size distribution and surface passivation
of the two fluorophores (Cao et al., 2022).
3.2. Morphology and structure of ratiometric fluorescent aerogels
The structural composition and content of the surface groups of
aerogels CF, CF-CD and CF-PpIX were analysed by XPS using split peak
fitting. The covalent grafting reaction equation of CDs and PpIX was
shown in Fig. 3a. The two fluorophores can be covalently grafted as
cross-linking agents, resulting in a tighter three-dimensional network
structure and giving the aerogel good mechanical properties (Wu, Zhu,
Dufresne, & Lin, 2019). As shown in Fig. 3b–e, Table S1, and Table S2,
the C1s fractionation of CF aerogels yielded C– – C/C–C/C–H, C–N,
O–– C–C, and C–O–C/C–H–C with bond energies and contents of 284.63
eV (36.65%), 285.43 eV (27.73%), 288 eV (18.86%), 286.51 eV
(32.58%) (Wang, Guo, Zhu, He, & Wang, 2021; Zhou, Zhu, Xue, He, &
Wang, 2020). CF-CD aerogels had an increased C–O–C/C–H–O content
compared to CF aerogels and the N1s fractionated C– – N double bond
appeared and was 23.52%, as shown in Fig. 3f–h, Table S2 (Song et al.,
2016). CF-PpIX aerogels had a C– – N content of 22.59% and an O– – C–C
content of 23.35%, which was much more than the O– – C–C content of
CF. This is mainly due to the condensation and esterification reaction of
CF-PpIX with CF (Pandele et al., 2020). The C (78.69%) and N (9.7%)
content of CF-CD and the O (16.96%) and N (13.7%) content of CF-PpIX
were significantly more than the CF elemental content based on the

Fig. 4. SEM and EDX spectra of CF (a, b, c, d), CF-CP (e, f, g, h).

4
S. Yang et al. Food Control 150 (2023) 109775

Fig. 5. Two-dimensional image of the CF-CP laser confocal microscope(CLSM) (a) CF-CD, (b) CF-PpIX, (c) CF-CP. Micro-CT image of CF-CP, (d) Front view, (e)
hollowing, (f) transverse section.

Fig. 6. (a) Fluorescence images of CF-CD and CF-

PpIX mixed in different ratios and corresponding
aerogels under UV light irradiation. (b) FTIR spectra
of CF, CF-CD, CF-PpIX. (c) XRD spectra of CF, CF-CD,
CF-PpIX, CF-CP. (d) UV–Vis spectra of CF-CD, CF-
PpIX. (e) Collagen-based dual-emission mixture so­
lutions with different CF-PpIX (red) to CF-CD (blue)
ratios (w/w) of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8,
1:9, 1:10, 1:11, and the corresponding lyophilized
aerogel solid fluorescent materials under 365 nm UV
lamp irradiation.

5
S. Yang et al.
analysis of the major single elements. The above was a good indication
of the successful introduction of both CDs and PpIX luminophores into
the CF structure.
It can be seen from SEM that the overall surface of pure CF and CF-CP
has a regular sheet-like structure with a size of about 50–100 μm and
high porosity as shown in Fig. 4a–h. The surface of the CF-CP is
smoother and appears flatter. This may be due to the strong and flat 3D
network structure of the aerogels due to the grafted CDs and PpIX acting
as cross-linking agents (Wu et al., 2019). From the corresponding EDX, it
can be seen that CF-CP contain higher O and N contents than CF aerogels
as shown in Fig. 4d, h. This also confirmed the results of grafting CDs and
PpIX on CF.
Blue and red fluorescence of CDs and PpIX on CF excited at 400 nm
by CLSM as shown in Fig. 5a–c. Simultaneous CLSM 2D images and
micro-CT analysis showed that both CDs and PpIX were present on the
lamellar fibers and evenly distributed on the CF emitting blue/red
fluorescence as shown in Fig. 5a–f. As in Fig. S2, the 3D CLSM map of the
CF-CP further exhibits the structure and uniform distribution of CF-CD
and CF-PpIX in the same system. The size and 3D network structure of
the CF-CP can be clearly determined from the fluorescence and are
consistent with the SEM images.
3.3. Detection of alkaline gases by specific ratiometric fluorescent aerogels
As shown in Fig. 6a, all three aerogel FT-IRs contained amide A, B, I,
II and III characteristic absorption peaks at 3307 cm− 1, 3080 cm− 1,
1657 cm− 1, 1545 cm− 1and 1245 cm− 1 respectively (Li et al., 2013; F. Liu
et al., 2017; X. H. Liu, Dan, & Dan, 2016). The absorption peak of the
6
Food Control 150 (2023) 109775
Fig. 7. (a) Fluorescence response of CF-CD to
different ammonia concentrations (0-2.0 × 10− 2 mg/
L). (b) Non-linear fit analysis of the fluorescence in­
tensity ratio of CF-CD to CF-PpIX (ICD/IPpIX) versus
ammonia concentration. (c) Fluorescence response of
CF-CD to various BAs (1.0 × 10− 3 mg/L). (d) Fluo­
rescence response of CF-CD in different concentra­
tions of dimethylamine solutions (0-2.0 × 10− 2 mg/
L). (e) Laboratory diffraction model analysis of CF-CP
solutions in different concentrations of BAs solutions.
(f) Red-green-blue (RGB) colour model analysis of CF-
CP placed with shrimp at − 4 ◦ C.
amide A bond at 3307 cm− 1 is mainly due to the stretching vibrations of
O–H and N–H, and the absorption peak of the amide B bond is attributed
to the asymmetric stretching vibrations of –CH2. The amide I and II
absorption peaks at 1657 cm− 1 and 1545 cm− 1 come from the stretching
vibration of the amide C–– O and the N–H bending vibration in CF’s own
protein structure, respectively (He et al., 2011). The two peaks at CF-CD
were higher than those at CF due to the condensation of –NH2 of CDs and
–COOH of CF to form the amide bond –CO–NH-. The CF-PpIX peak was
significantly higher than that of CF and CF-CD, which was attributed to
the esterification of –NH2 and –OH of CF with –COOH of PpIX. The C–N
stretching vibrations of the amide III bond at 1245 cm− 1 and the N–H
bending vibrations at 1245 cm− 1 produce absorption peaks from both
the fluorophore’s own structure and the covalently bound amide bond
structure, and both have peaks higher than CF. This was consistent with
the analysis results of XPS.
The changes in the crystal structure of CF after covalent grafting
were analysed using X-ray diffraction, as shown in Fig. 6b. As can be
seen from the graphs, the peak positions and peak intensities did not
change significantly, indicating that the covalent grafting did not change
the crystalline structure of CF. The effect of the grafted CF-CD and CF-
PpIX on the thermal stability of the original CF was analysed by ther­
mogravimetry. As shown in Fig. S3 (a, b) each curve showed two stages
of mass loss at 20◦ C–200 ◦ C and 200◦ C–600 ◦ C respectively. This is
mainly attributed to the formation of different covalent bonds between
the molecular chains during CF grafting, changing the hydrogen bond
content between the collagen chains and the peptide chain moieties.
Overall, there was no significant difference in the thermal stability of CF-
CP and CF after covalent grafting and mixing. When the CF-CP is placed
S. Yang et al.
Fig. 8. The freshness of shrimps (Approx. 14 g) was monitored in real time using CF-CP at different temperatures (− 18 ◦ C, 4 ◦ C, 25 ◦ C).
in water, it immediately absorbs water and swells, disrupting the three-
dimensional mesh structure of the aerogel, and for a minute the CF-CP is
completely fused with the water system, as in Supplementary Video 1.
This is probably due to the fact that the molecular structure of collagen
fibers contains a large number of hydrophilic reactive groups such as
–COOH, –NH2 and –OH, which are not only involved in chemical
modifications but also hydrogen-bonded to water molecules, leading to
their enhanced hydrophilic capacity (Song et al., 2016). After the CF-CP
was fused with water and left for 24 h, the solution did not change
significantly, as shown in fig. S4. This reflects the good hydrophilicity
and stability of CF-CP.
Supplementary video related to this article can be found at https://
doi.org/10.1016/j.foodcont.2023.109775.
As shown in Fig. 6c, the CF-CD UV absorption peak at 279 nm and the
CF-PpIX UV absorption peak at 394 nm are responsible for the blue and
red fluorescence, respectively. The excitation wavelengths of blue-
emitting CF-CD and red-emitting CF-PpIX partially overlap, indicating
that CF-CD and CF-PpIX can be excited by the same wavelength at
different locations in the 358–475 nm range, producing two different
colors of fluorescence, as shown in Fig. 6d (Jia et al., 2019). The colour
change from red to blue was presented by adjusting different ratios of
CF-CD to CF-PpIX. The subsequent experimental investigation was car­
ried out by selecting the optimum ratio of CF-CD: CF-PpIX = 10:1, as
shown in Fig. 6e.
3.4. Detection of alkaline gases by specific ratiometric fluorescent aerogels
The variation of ammonia on the fluorescence spectrum was
explored with a mixture of fluorescent materials at a ratio of CF-CD:CF-
PpIX = 10:1. As shown in Fig. 7a, the two characteristic emission peaks
of CF-CD and CF-PpIX were represented at 460 nm and 636 nm
respectively. The fluorescence intensity of CF-CD at 460 nm decreased
with increasing ammonia concentration after mixing the ratio mixture
solution with ammonia, and the fluorescence intensity of CF-PpIX at
636 nm was not significantly affected by the ammonia concentration.
This is the same as that shown in Fig. 2b, e. The alkaline solution
decreased the fluorescence intensity of CF-CD and had no effect on CF-
PpIX, and none of the peak positions changed. The fluorescence intensity
of the CDs at 460 nm decreased with increasing ammonia concentration
and was further fitted ratiometrically and non-linearly to a background
PpIX as in Fig. 7b. Based on the results of the fit, the curve can be used as
a function y = 1.67486 + 0.27807e-x/50.87305 + 0.57003e-x/
6197.4906 (R2 = 0.9813) for the description. The detection limit for BAs
can be as low as 1.0 × 10− 6 mg/L for this ratio of fluorescent aerogel,
demonstrating the ease of operation, low cost and relatively low
detection limits of fluorescence detection compared to other detection
methods, as shown in Table S4.
As shown in Fig. 7c, CF-CP reacted with all selected VOCs and
showed good selectivity. Of these, dimethylamine had the greatest effect
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Food Control 150 (2023) 109775
on CF-CP and tyramine the least, reflecting the fact that CF-CP has better
fold selection. It was confirmed that bases change the structure of CDs
and thus their fluorescence intensity. In the case of dimethylamine, the
fluorescence intensity of CF-CP at 460 nm decreased gradually with
increasing concentration of dimethylamine (0-2.0 × 10− 2 mg/L), which
was the same trend as the fluorescence intensity change curve at 460 nm
for CF-CP with ammonia.
In BAs gas, the proportional fluorescent aerogel can not only be
visualised as a change in colour from blue to deep red, but the stability of
the colour can also be determined from Lab and RGB values (Ge et al.,
2019; J. J. Zhang et al., 2019). As shown in Fig. 7e, different concen­
trations of ammonia produced significant colour changes and corre­
sponding lab values for the mixed solutions under 365 nm UV lamp
irradiation. The results showed an increasing trend for both red to green
(a) and yellow to blue (b) deviation values and the opposite for L
brightness values. The total colour difference (ΔE) for both orders of
magnitude of the CF-CP mixture was greater than 8, exceeding the
minimum value of 3 for the total colour difference that can be distin­
guished by the human eye. This also demonstrated that real-time
monitoring by the naked eye can be achieved. To illustrate the sensi­
tivity and stability of the CF-CP during the detection process, the ac­
curate correspondence of the results was determined by the colour
change of fresh shrimps at 4 ◦ C over a period of one week and the cor­
responding RGB values in response. As shown in Fig. 7f, the blue value
(B) of CF-CP compared with fluorescent aerogels coexisting with fresh
shrimp under 365 nm ultraviolet light decreased to 214, the green value
(G) decreased by 39.33%, and the red value increased slightly. The
comprehensive value of the three primary colors SRGB increased to
26.68% and the average value was also greater than 12.82%. CF-CP offer
more outstanding fluorescence changes and stability. The CF-CP aerogel
showed no significant change in its fluorescence intensity within a week
and had an anti-photobleaching effect, as shown in Fig. S5. This also
reinforces that CF-CP can be used as a intelligent label for real-time
detection in food spoilage and has a wide potential for commercial
application.
3.5. CF-CP ratiometric fluorescent aerogels for food detection
As shown in Fig. 8, under the 365nm UV lamp, we observed with the
naked eye that over time the CF-CP in cling film dishes at 25 ◦ C and 4 ◦ C
was initially blue, then the blue disappeared to red, followed by a
gradual increase in red colour, which also indicated that the fresh
shrimp were slowly decaying. At 25 ◦ C, the aerogel changes colour to
plum after 2 days, indicating complete spoilage of the food. At 4 ◦ C the
aerogel changed from blue to red after 3 days and after 5 days the aer­
ogel changed completely to red with a subsequent deepening of the red
colour, indicating that the shelf life of the food could be extended at low
temperatures. At frozen − 18 ◦ C, the ratio fluorescence aerogel showed
no visible colour change, indicating that fresh shrimps can be stored for
S. Yang et al.
Fig. 9. (a) FTIR spectra of CDs and CDs after reaction with ammonia. (b) Wide-scan XPS of CF-CP before and after adsorption of shrimp spoilage gas.(c) Decon­
voluted peaks of the N1s XPS spectrum of CF-CP befor reaction,(d) CF-CP after reaction.
a week or more under frozen conditions without deterioration and
spoilage. The CF-CP can be used as a intelligent label for food spoilage
detection.
3.6. Mechanism of CF-CP ratiometric fluorescent aerogel response to
basic gases
The mechanism of the interaction of alkalis with CF-CP was revealed
by using XPS for CF-CD aerogels before and after the biogenic amine gas
reaction and FTIR for the solution of CDs before and after the ammonia
water reaction for group structure analysis. As shown in Fig. 9a, the
stretching vibrations at 3446 cm− 1 were attributed to –OH and –NH, and
1646 cm− 1 and 1564 cm− 1 were generated by the stretching vibrations
of C–– N, C–
– O and the bending vibrations of –NH, respectively (He et al.,
2011; Song et al., 2016). In particular, a transition from a sharp to a
gentle peak occurs at 3446 cm− 1 compared to CDs, with a significantly
weaker C– – N peak at 1646 cm− 1 and an enhanced –NH peak at 1564
− 1
cm . This may be due to the conversion of the C– – N to C–NH2 group on
the CDs (Song et al., 2016). To further confirm this hypothesis, the N1s
were fitted by XPS to the sub-peaks and the corresponding content data
were analysed as shown in Fig. 9c.d, Table S3. The C– – N, –NH and
-N-(C)3 contents of the CDs were 52.33%, 3.26% and 44.41%, respec­
tively. In comparison with the CDs, the C– – N (39.46%) content of the
reacted CDs decreased by 24.59%, the –NH (8.1%) content increased by
4.84% and the -N-(C)3 (52.44%) content increased by 18.08%. There­
fore, the main reason for the change in fluorescence intensity of CDs
under alkaline conditions may be due to the conversion of some of the
C–– N groups to -C-NH2 groups in the structure (Song et al., 2016).
4. Conclusion
Overall, CF-CP exhibits a remarkable response to BAs, with detection
limits as low as 1.0 × 10− 6 mg/L for BAs, overcoming the poor sensi­
tivity of most gas sensors. The blue-emitting CDs (pH indicator) and the
red-emitting PpIX (background reference) were mixed in different ra­
tios, the colour was changed from blue to deep red and the final ratio of
8
Food Control 150 (2023) 109775
10:1 was optimal for the experiment. As the concentration of BAs
increased, the colour of CF-CP changed significantly, from blue to dark
red, which was consistent with the results of the fresh shrimp spoilage
assay. This may be due to the weakening of the blue fluorescence caused
by the conversion of CDs to C–– N upon encountering the basic substance
C–N. The analysis of SRGB and ΔE values shows that CF-CP has a high
degree of confidence as a intelligent label. In addition, the CF-CP based
on collagen fibers possesses a large number of hydrophilic groups, such
as –NH2, –COOH and –OH groups that can absorb water, swell and
dissolve in 1 minute upon contact with water, and this rapid solubility
also provides a reference for other research directions. The development
and application of multifunctional biomass-based materials can thus be
of great importance in terms of food monitoring, promoting the practical
application and development of the food industry.
CRediT authorship contribution statement
Shuo Yang: Supervision. Jiang Lou: Visualization, Investigation.
Limin Jing: Writing – original draft, Writing – review & editing, and.
Qijun Ding: Software. Xia Li: All authors contributed as the main
contributors of this work. Yifei Jiang: Data curation, Writing – original
draft. Zhuqing Liu: Conceptualization, Methodology. Wenjia Han:
Supervision.
Declaration of interest statement
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
S. Yang et al.
Acknowledgements
This work is supported by Foundation of the Shandong Key R&D
Program (No.2021CXGC011002), the Natural Science Foundation of
Shandong Province, China (No. ZR2021QB009 & No. ZR2021QC158),
the QUTJBZ Program (No. 2022JBZ01-05), and Outstanding Youth
Innovation Team Project of Shandong Provincial University
(242007040109).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.foodcont.2023.109775.
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