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Nanomaterial Based Emerging Technologies for
Detecting Food Contaminants 1st Edition Mingfei Pan
Digital Instant Download
Author(s): Mingfei Pan, Longhua Xu, Huilin Liu
ISBN(s): 9783725805952, 3725805954
Edition: 1
File Details: PDF, 17.07 MB
Year: 2024
Language: english
Special Issue Reprint

Nanomaterial-Based
Emerging Technologies
for Detecting Food
Contaminants

Edited by
Mingfei Pan, Longhua Xu and Huilin Liu

mdpi.com/journal/foods
Nanomaterial-Based Emerging
Technologies for Detecting Food
Contaminants
Nanomaterial-Based Emerging
Technologies for Detecting Food
Contaminants

Editors
Mingfei Pan
Longhua Xu
Huilin Liu

Basel • Beijing • Wuhan • Barcelona • Belgrade • Novi Sad • Cluj • Manchester

Editors
Mingfei Pan Longhua Xu Huilin Liu
Tianjin University of Science Shandong Agricultural Beijing Technology and
and Technology University Business University
Tianjin Taian Beijing
China China China

Editorial Office
MDPI
St. Alban-Anlage 66
4052 Basel, Switzerland

This is a reprint of articles from the Special Issue published online in the open access journal
Foods (ISSN 2304-8158) (available at: https://www.mdpi.com/journal/foods/special issues/
nanomaterial food contaminants).

For citation purposes, cite each article independently as indicated on the article page online and as
indicated below:

Lastname, A.A.; Lastname, B.B. Article Title. Journal Name Year, Volume Number, Page Range.

ISBN 978-3-7258-0595-2 (Hbk)

ISBN 978-3-7258-0596-9 (PDF)
doi.org/10.3390/books978-3-7258-0596-9

© 2024 by the authors. Articles in this book are Open Access and distributed under the Creative
Commons Attribution (CC BY) license. The book as a whole is distributed by MDPI under the terms
and conditions of the Creative Commons Attribution-NonCommercial-NoDerivs (CC BY-NC-ND)
license.
Contents

Mingfei Pan
Nanomaterial-Based Optical Detection of Food Contaminants
Reprinted from: Foods 2024, 13, 557, doi:10.3390/foods13040557 . . . . . . . . . . . . . . . . . . . 1

Tianyu Ma, Kaixin Liu, Xiao Yang, Jingying Yang, Mingfei Pan and Shuo Wang
Development of Indirect Competitive ELISA and Visualized Multicolor ELISA Based on Gold
Nanorods Growth for the Determination of Zearalenone
Reprinted from: Foods 2021, 10, 2654, doi:10.3390/foods10112654 . . . . . . . . . . . . . . . . . . 6

Lingyan Zhao, Jingyi Jin, Wenbo Zhu, Yuehua Zuo and Yang Song
Detection of Pyrethroids in Food by Immunofluorescence Enhanced Method Based on
Three-Layer Core-Shell Structure Upconversion Materials
Reprinted from: Foods 2022, 11, 990, doi:10.3390/foods11070990 . . . . . . . . . . . . . . . . . . . 18

Wenbo Zhu, Lingyan Zhao, Jingyi Jin and Yang Song

Preparation of Core-Shell Rare Earth-Doped Upconversion Nanomaterials and Simultaneous
Detection of Two Pesticides in Food
Reprinted from: Foods 2022, 11, 1485, doi:10.3390/foods11101485 . . . . . . . . . . . . . . . . . . 32

Chang Liu, Haiyang Wang, Xuelian Hu, Yichuan Cao and Guozhen Fang
Construction of an ECL Detection Platform for Sensitive Detection of Carbaryl Based on an
Eu3+ -Functionalized Metal–Organic Framework Encapsulated with Nanogold
Reprinted from: Foods 2022, 11, 1487, doi:10.3390/foods11101487 . . . . . . . . . . . . . . . . . . 48

Liping
Hong,
Mingfei
Pan,
Xiaoqian
Xie,
Kaixin
Liu,
Jingying
Yang,
Shan
Wang
and

Shuo
Wang
Aptamer-Based
Fluorescent
Biosensor
for
the
Rapid
and
Sensitive
Detection
of
Allergens
in

Food
Matrices
Reprinted from: Foods 2021, 10, 2598, doi:10.3390/foods10112598 . . . . . . . . . . . . . . . . . . 61

Ziwen
Zhang,
Ran
Han,
Sixuan
Chen,
Feilin
Zheng,
Xinmiao
Ma,
Mingfei
Pan
and
Shuo
Wang
Fluorescent
and
Colorimetric
Dual-Mode
Strategy
Based
on
Rhodamine
6G
Hydrazide
for

Qualitative
and
Quantitative
Detection
of
Hg2+
in
Seafoods
Reprinted from: Foods 2023, 12, 1085, doi:10.3390/foods12051085 . . . . . . . . . . . . . . . . . . 82

Shijie Li, Linqing Nie, Lin Han, Wenjun Wen, Junping Wang and Shuo Wang
Glutathione-Capped CdTe Quantum Dots Based Sensors for Detection of H2 O2 and
Enrofloxacin in Foods Samples
Reprinted from: Foods 2023, 12, 62, doi:10.3390/foods12010062 . . . . . . . . . . . . . . . . . . . . 92

Qi Zhang, Haiyang Wang, Yongju Zhang, Zhixiang Xu and Longhua Xu

Preparation of Magnetic Metal-Organic Frameworks@Molecularly Imprinted Nanoparticles for
Specific Extraction and Enrichment of Bisphenol A in Food
Reprinted from: Foods 2022, 11, 1408, doi:10.3390/foods11101408 . . . . . . . . . . . . . . . . . . 108

Ying Guo, Guanqing Yuan, Xuelian Hu, Jinni Zhang and Guozhen Fang
A High-Luminescence Biomimetic Nanosensor Based on N, S-GQDs-Embedded Zinc-Based
Metal–Organic Framework@Molecularly Imprinted Polymer for Sensitive Detection of
Octopamine in Fermented Foods
Reprinted from: Foods 2022, 11, 1348, doi:10.3390/foods11091348 . . . . . . . . . . . . . . . . . . 120

v
Xiaohui Wang, Chang Liu, Yichuan Cao, Lin Cai, Haiyang Wang and Guozhen Fang
A Turn-Off Fluorescent Biomimetic Sensor Based on A Molecularly Imprinted Polymer-Coated
Amino-Functionalized Zirconium (IV) Metal–Organic Framework for the Ultrasensitive and
Selective Detection of Trace Oxytetracycline in Milk
Reprinted from: Foods 2023, 12, 2255, doi:10.3390/foods12112255 . . . . . . . . . . . . . . . . . . 134

vi
 foods
Editorial
Nanomaterial-Based Optical Detection of Food Contaminants
Mingfei Pan

State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology,
Tianjin 300457, China; [email protected]

1. Introduction
The presence of food contaminants remains a significant aspect contributing to global
food safety issues, drawing widespread attention from ordinary consumers, governments,
and researchers [1–3]. These contaminants encompass various harmful factors, including
residues of agricultural and veterinary drugs, biotoxins, heavy metals, allergenic pro-
teins, and particularly endogenous hazardous substances generated during processing,
representing a new focus in detection technology research [4,5].
Various nanomaterials with different structures and properties, such as metallic nano-
materials, up-conversion fluorescence nanomaterials, metal–organic framework porous
materials, quantum dot fluorescence materials, etc., not only serve as solid carriers for bio-
recognition elements (such as antibodies, aptamers, etc.) [6,7] and biomimetic recognition
elements (such as molecularly imprinted polymers) [8,9], but also provide signal sources for
visual, rapid, and convenient analysis [10–12]. Visible light, fluorescence, or electrochemi-
luminescence are quantitative signals commonly used in convenient food safety analysis
strategies. This significantly promotes the development of precise and rapid analysis
techniques for food contaminants, incorporating advanced methods such as nanomaterials,
biomimetics and biorecognition, and chemometrics. Therefore, strategies based on mul-
tifunctional nanomaterials and utilizing antibodies, aptamers, and biomimetic polymers
as recognition elements for the fluorescent or visual detection of contaminants in complex
food matrices are gradually assuming crucial roles in food safety testing strategies [13–15].

2. An Overview of Published Articles

In the study by Tianyu Ma et al. (Contribution 1), the authors designed and prepared
Citation: Pan, M. a zearalenone (ZEN) hapten against the mycotoxin ZEN, and the original coating ZEN-
Nanomaterial-Based Optical ovalbumin (ZEN-OVA) by conjugation with OVA. Based on gold nanorods (AuNRs) of
Detection of Food Contaminants. uniform size and stable properties synthesized by the seed-mediated method, the indirect
Foods 2024, 13, 557. competitive enzyme-linked immunosorbent assay (ic-ELISA) and the AuNR growth-based
https://doi.org/10.3390/ multicolor ELISA for detecting ZEN toxin were further established. Under optimal exper-
foods13040557 imental conditions, the coating amounts of ZEN-OVA were 0.025 μg/well, the antibody
Received: 26 January 2024 (Ab) dilution factor was 32,000 times, blocking solution was 0.5% skimmed milk powder,
Accepted: 3 February 2024 enzyme-labeled secondary Ab diluted 10,000 times, and at pH 7.4 of the PBS buffer, the
Published: 12 February 2024 sensitivity (IC50 ) of the established ic-ELISA for ZEN detection reached 0.85 ± 0.04 μg/L
and the limit of detection (LOD, IC15 ) reached 0.22 ± 0.08 μg/L. In the multicolor ELISA
based on the growth of AuNRs, as the content of ZEN increased, the mixed solution ex-
hibited a significant color change from brownish red to colorless. ZEN concentrations as
Copyright: © 2024 by the author. low as 0.1 μg/L could be detected with the naked eye (brown-red to dark gray). This study
Licensee MDPI, Basel, Switzerland. presents an effective analysis strategy for the rapid screening and accurate monitoring of
This article is an open access article
ZEN contaminants in foods.
distributed under the terms and
In the study by Lingyan Zhao et al. (Contribution 2), a novel rare earth upconversion
conditions of the Creative Commons
nanomaterial with a three-layer sandwich core–shell structure was synthesized by an im-
Attribution (CC BY) license (https://
proved thermal decomposition method, and the morphology, fluorescence intensity, and
creativecommons.org/licenses/by/
diffraction peak position of the new material were characterized by TEM (transmission
4.0/).

Foods 2024, 13, 557. https://doi.org/10.3390/foods13040557 https://www.mdpi.com/journal/foods

1
Foods 2024, 13, 557

electron microscopy), XRD (powder X-ray diffraction), and fluorescence spectrophotome-

try. The inert core/active shell/inert shell design improved the upconversion luminous
efficiency of the new material several-fold. FT-IR (Fourier transform infrared spectroscopy)
characterization showed that the surface of activated upconversion nanoparticles was
modified with the silicon shell and amino group. Combined with the characteristics that
aminoated polystyrene magnetic microspheres could be separated by the magnetic field,
an upconversion magnetic separation immunoassay method for the detection of pyrethroid
pesticide residues was established. The capture probe competed with the pyrethroid stan-
dard, combined the signal probe, and measured the fluorescence signal value formed by
the capture probe signal probe complex at 542 nm under 980 nm excitation light. The
LOD of fenpropathrin was 0.01 μg/L, cypermethrin was 0.015 μg/L, and fenvalerate was
0.011 μg/L. Through the actual detection of apple, cabbage, and other samples, the re-
covery rate of pyrethroids was between approximately 83.4% and 97.8%. Comparison
with the HPLC (high-performance liquid chromatography) detection results showed that
the established method had good accuracy and could realize the quantitative analysis of
pyrethroids in food.
In their research, Wenbo Zhu et al. (Contribution 3) coupled two upconversion
materials with anti-clothianidin and anti-imidacloprid monoclonal antibodies as signal
probes using the glutaraldehyde cross-linking method. Under the excitation of 980 nm
excitation light, the fluorescence signals of the synthesized core–shell NaYF4 :Yb@NaYF4 :Ho
and monolayer NaYF4 :Yb,Tm upconversion nanoparticles (UCNPs) were simultaneously
detected at 656 and 696 nm, respectively. Imidacloprid (IMI) and clothianidin (CLO) could
compete with antigen-conjugated amino Fe3 O4 magnetic nanomaterials for binding to
signaling probes, thus establishing a rapid and sensitive fluorescent immunoassay for the
simultaneous detection of IMI and CLO. Under optimal conditions, the LOD (IC10 ) and
sensitivity (IC50 ) of IMI and CLO were (0.032, 0.028) and (4.7, 2.1) ng/mL, respectively, and
the linear assay ranges were at 0.032–285.75 ng/mL and 0.028–200 ng/mL, respectively. The
immunoassay did not significantly cross-react with other analogs. In fruits and vegetables
such as apples, oranges, peaches, cucumbers, tomatoes, and peppers, the mean recoveries
of IMI and CLO ranged from 83.33% to 115.02% with relative standard deviations (RSDs) of
1.9% to 9.2% and 1.2% to 9.0%, respectively. Furthermore, the results of the immunoassay
correlate well with the high-performance liquid chromatography method used to detect
the actual samples.
In the work of Chang Liu et al. (Contribution 4), a Eu3+ -MOF-253@Au electrochemilu-
minescence sensor was successfully constructed for the first time by encapsulating nanogold
in the metal–organic framework (MOF) backbone and pore channels, and assembling Eu3+
on the MOF backbone. Firstly, the introduction of nanogold overcame the weakness of
MOFs, which was difficult to achieve, and enhanced its catalytic performance, followed
by the modification of Eu3+ to confer the electrochemiluminescence performance and the
function of target detection on the sensor. Moreover, carbaryl was placed in an alkaline
working solution to enhance the intensity of electrochemiluminescence signals, as well
as to promote the hydrolysis of carbaryl into 1-naphthol, which caused the burst of the
Eu3+ -MOF-253@Au electrochemiluminescence sensor, thereby achieving the sensitive de-
tection of carbaryl. On this basis, the electrochemiluminescence detection conditions were
optimized, the performance was analyzed, and finally, it was successfully used for the
detection of carbaryl with good linearity in the range of 0.2–200 μg L−1 and a low LOD
(0.14 μg L−1 ).
Food allergies have seriously affected some people’s quality of life, and even endan-
gered their lives. At present, there is still no effective cure for food allergies. Avoiding the
intake of allergenic food is still the most effective way to prevent allergic diseases. Therefore,
it is necessary to develop rapid, accurate, sensitive, and reliable analysis methods to detect
food allergens from different sources. Aptamers are oligonucleotide sequences that can
bind to a variety of targets with high specificity and selectivity, and they are often combined
with different transduction technologies, thereby constructing various types of aptamer

2
Foods 2024, 13, 557

sensors. In recent years, with the development of technology and the application of new
materials, the sensitivity, portability, and cost of fluorescence sensing technology have been
greatly improved. Therefore, aptamer-based fluorescence sensing technology has been
widely developed and applied in the specific recognition of food allergens. Liping Hong
et al. (Contribution 5) comprehensively reviewed the classification of major allergens and
their characteristics in animal and plant foods, and summarized the preparation principles
and practical applications of aptamer-based fluorescence biosensors. This article presents
some strategies for the rapid and sensitive detection of allergens in food matrices.
In their study, Ziwen Zhang et al. (Contribution 6) developed a rapid fluorescent and
colorimetric dual-mode detection strategy for Hg2+ in seafoods based on the cyclic binding
of the organic fluorescent dye rhodamine 6G hydrazide (R6GH) to Hg2+ . The luminescence
properties of the fluorescent R6GH probe in different systems were investigated in detail.
Based on the UV and fluorescence spectra, it was determined that the R6GH has good
fluorescence intensity in acetonitrile and good selective recognition of Hg2+ . Under optimal
conditions, the R6GH fluorescent probe showed a good linear response to Hg2+ (R2 = 0.9888)
in the range of 0–5 μM, with a low detection limit of 2.5 × 10−2 μM (S/N = 3). A paper-
based sensing strategy based on fluorescence and colorimetric analysis was developed for
the visualization and semiquantitative analysis of Hg2+ in seafoods. The LAB values of
the paper-based sensor impregnated with the R6GH probe solution showed good linearity
(R2 = 0.9875), with Hg2+ concentrations in the range of 0–50 μM, which means that the
sensing paper could be combined with smart devices to provide reliable and efficient
Hg2+ detection.
Additives and antibiotic abuse during food production and processing are among the
key factors affecting food safety. The efficient and rapid detection of hazardous substances
in food is of crucial relevance to ensure food safety. In the study by Shijie Li et al. (Con-
tribution 7), a water-soluble quantum dot with glutathione as a ligand was synthesized
as a fluorescent probe by the hydrothermal method to achieve the detection and analysis
of H2 O2 . The detection limits were 0.61 μM in water and 68 μM in milk. Moreover, it
was used as a fluorescent donor probe, and manganese dioxide nanosheets were used as
a fluorescent acceptor probe in combination with an immunoassay platform to achieve
the rapid detection and analysis of enrofloxacin (ENR) in a variety of foods with LODs of
0.05–0.25 ng/mL in foods. The proposed systems provide new ideas for the construction of
fluorescence sensors with high sensitivity.
The study from Qi Zhang et al. (Contribution 8) proposes a facile and versatile
layer-by-layer strategy without any special surface modifications for the preparation of
magnetic metal-organic frameworks (MMOFs) supporting molecularly imprinted poly-
mer nanoparticles (MMOFs@MIP), which are based on a magnetically susceptible core
conjugated with an imidazole-derived self-assembled layer and a silane-based imprinted
shell. Metal–organic frameworks (MOFs) with systematically tailored structures have been
suggested as promising precursors to the preparation of diverse functional materials. The
obtained MMOFs@MIPs, which integrated the advantages of Fe3 O4 , MOFs, and MIPs,
were characterized and exhibited good magnetic properties, a rapid mass transfer rate, and
excellent adsorption selectivity, as well as capacity for the targeted molecular—bisphenol A
(BPA). Moreover, the MMOFs@MIPs were employed as adsorbents in magnetic solid-phase
extraction (MSPE) to selectively bind and rapidly separate BPA from real samples, with
satisfactory recovery rates ranging from 88.3% to 92.3%. More importantly, the desirable
reusability of MMOFs@MIPs was also evaluated, and the recovery was maintained above
88.0% even after five re-use cycles. Furthermore, combined with high-performance liquid
chromatography (HPLC) analysis, a novel MSPE-HPLC method was developed, enabling
the highly selective and sensitive detection of BPA in a wide linear range of 0.5–5000 μg L−1 ,
with a low LOD of 0.1 μg L−1 . This work contributes a promising method for constructing
various functional nanoparticle@MOFs@MIP hybrid materials for applications in many
different fields.

3
Foods 2024, 13, 557

Ying Guo et al. constructed a novel fluorescent molecularly imprinted nanosensor

(N, S-GQDs@ZIF-8@MIP) based on the nitrogen and sulfur co-doped graphene quantum
dots decorated zeolitic imidazolate framework-8 for the detection of octopamine (OA)
(Contribution 9). Herein, ZIF-8 with a large surface area was introduced as a supporter of
the sensing system, which effectively shortened the response time of the sensor. Meanwhile,
high green luminescent N, S-GQDs and a maximum emission wavelength of 520 nm
under 460 nm excitation and a 12.5% quantum yield, were modified on the surface of
ZIF-8 as a signal tag that could convert the interactions between the sensor and OA into
detectable fluorescent signals. Finally, N, S-GQDs@ZIF-8@MIP was acquired through the
surface molecular imprinting method. Due to the synergy of N, S-GQDs, ZIF-8, and MIP,
the obtained sensor not only demonstrated higher selectivity and sensitivity than N, S-
GQDs@ZIF-8@NIP, but also displayed faster fluorescence responses than N, S-GQDs@MIP.
Under optimal conditions, the developed sensor presented a favorable linear relationship
in the range of 0.1–10 mg L−1 , with a detection limit of 0.062 mg L−1 . Additionally, the
proposed N, S-GQDs@ZIF-8@MIP strategy was effectively applied to the detection of OA
in fermented samples.
Xiaohui Wang et al. constructed a fluorescent sensor (NH2 -UIO-66(Zr)@MIP) based
on a molecularly imprinted polymer-coated amino-functionalized zirconium (IV) metal–
organic framework, and initially used it for the ultrasensitive determination of oxytetracy-
cline (Contribution 10). Developing sensitive and effective methods to monitor oxytetracy-
cline residues in food is of great significance for maintaining public health. NH2 -UIO-66
(Zr), with a maximum emission wavelength of 455 nm under 350 nm excitation, was
prepared using a microwave-assisted heating method. The NH2 -UIO-66(Zr)@MIP sen-
sor with specific recognition sites for oxytetracycline was then acquired by modifying a
molecularly imprinted polymer on the surface of NH2 -UIO-66 (Zr). The introduction of
NH2 -UIO-66 (Zr) as both a signal tag and supporter could strengthen the sensitivity of the
fluorescence sensor. Thanks to the combination of the unique characteristics of the molecu-
larly imprinted polymer and NH2 -UIO-66 (Zr), the prepared sensor not only exhibited a
sensitive fluorescence response, specific identification capabilities, and a high selectivity
for oxytetracycline, but also showed good fluorescence stability, satisfactory precision, and
reproducibility. The fabricated sensor displayed fluorescent linear quenching in the OTC
concentration range of 0.05–40 μg mL−1 , with a detection limit of 0.012 μg mL−1 . More
importantly, the fluorescence sensor was finally applied for the detection of oxytetracycline
in milk, and the results were comparable with those obtained using the HPLC approach.
Hence, the NH2 -UIO-66 (Zr)@MIP sensor possesses great application potential for the
accurate evaluation of trace oxytetracycline in dairy products.
Currently, detection technologies regarding food contaminants are still facing multiple
challenges, which include issues such as the effective removal of food matrices and the
monitoring and control of hazardous substances generated during food processing. It is
believed that the development of more efficient, accurate, and sensitive strategies for the
detection and control of food contaminants will be gradually promoted with the continuous
advancement of nanomaterials, food-related research, and chemometrics. The aim of this
Special Issue is to publish high-quality articles on the accurate and rapid detection of
various types of food contaminants in the areas of fluorescent nanomaterials, biometrics
and biomimetic recognition, and fluorescent molecules based on organic dyes and quantum
dots, in order to promote the further development of nanomaterial-based optical sensing
and detection methods.

Conflicts of Interest: The author declares no conflicts of interest.

References
1. Fung, F.; Wang, H.S.; Menon, S. Food safety in the 21st century. Biomed. J. 2018, 41, 88–95. [CrossRef] [PubMed]
2. Gallo, M.; Ferrara, L.; Calogero, A.; Montesano, D.; Naviglio, D. Relationships between food and diseases: What to know to
ensure food safety. Food Res. Int. 2020, 137, 109414. [CrossRef] [PubMed]

4
Foods 2024, 13, 557

3. Shen, Y.; Wei, Y.; Zhu, C.; Cao, J.; Han, D.M. Ratiometric fluorescent signals-driven smartphone-based portable sensors for onsite
visual detection of food contaminants. Coordin. Chem. Rev. 2022, 458, 214442. [CrossRef]
4. World Health Organization. Safety Evaluation of Certain Contaminants in Food: Prepared by the Ninetieth Meeting of the Joint FAO/WHO
Expert Committee on Food Additives (JECFA); WHO: Geneva, Switzerland, 2023.
5. Chen, J.; Sun, R.; Pan, C.; Sun, Y.; Mai, B.; Li, Q.X. Antibiotics and food safety in aquaculture. J. Agric. Food Chem. 2020, 68,
11908–11919. [CrossRef] [PubMed]
6. Ma, T.; Liu, K.; Yang, X.; Yang, J.; Pan, M.; Wang, S. Development of indirect competitive ELISA and visualized multicolor ELISA
based on gold nanorods growth for the determination of zearalenone. Foods 2021, 10, 2654. [CrossRef] [PubMed]
7. Hong, L.; Pan, M.; Xie, X.; Liu, K.; Yang, J.; Wang, S.; Wang, S. Aptamer-based fluorescent biosensor for the rapid and sensitive
detection of allergens in food matrices. Foods 2021, 10, 2598. [CrossRef] [PubMed]
8. Villa, C.C.; Sánchez, L.T.; Valencia, G.A.; Ahmed, S.; Gutiérrez, T.J. Molecularly imprinted polymers for food applications: A
review. Trends Food Sci. Technol. 2021, 111, 642–669. [CrossRef]
9. Ayerdurai, V.; Cieplak, M.; Kutner, W. Molecularly imprinted polymer-based electrochemical sensors for food contaminants
determination. TrAC-Trend. Anal. Chem. 2023, 158, 116830. [CrossRef]
10. Hua, Z.; Yu, T.; Liu, D.; Xianyu, Y. Recent advances in gold nanoparticles-based biosensors for food safety detection. Biosens.
Bioelectron. 2021, 179, 113076. [CrossRef] [PubMed]
11. Umapathi, R.; Park, B.; Sonwal, S.; Rani, G.M.; Cho, Y.; Huh, Y.S. Advances in optical-sensing strategies for the on-site detection
of pesticides in agricultural foods. Trends Food Sci. Technol. 2022, 119, 69–89. [CrossRef]
12. Zhang, Z.; Lou, Y.; Guo, C.; Jia, Q.; Song, Y.; Tian, J.Y.; Zhang, S.; Wang, M.; He, L.; Du, M. Metal–organic frameworks (MOFs)
based chemosensors/biosensors for analysis of food contaminants. Trends Food Sci. Technol. 2021, 118, 569–588. [CrossRef]
13. Umapathi, R.; Sonwal, S.; Lee, M.J.; Rani, G.M.; Lee, E.S.; Jeon, T.J.; Kang, S.M.; Oh, M.H.; Huh, Y.S. Colorimetric based on-site
sensing strategies for the rapid detection of pesticides in agricultural foods: New horizons, perspectives, and challenges. Coordin.
Chem. Rev. 2021, 446, 214061. [CrossRef]
14. Nile, S.H.; Baskar, V.; Selvaraj, D.; Nile, A.; Xiao, J.; Kai, G. Nanotechnologies in food science: Applications, recent trends, and
future perspectives. Nano-Micro Lett. 2020, 12, 45. [CrossRef] [PubMed]
15. Barzegar, F.; Kamankesh, M.; Mohammadi, A. Recent development in formation, toxic effects, human health and analytical
techniques of food contaminants. Food Rev. Int. 2023, 39, 1157–1183. [CrossRef]

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5
 foods
Article
Development of Indirect Competitive ELISA and Visualized
Multicolor ELISA Based on Gold Nanorods Growth for the
Determination of Zearalenone
Tianyu Ma 1,2 , Kaixin Liu 1,2 , Xiao Yang 1,2 , Jingying Yang 1,2 , Mingfei Pan 1,2, * and Shuo Wang 1,2

1 State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology,
Tianjin 300457, China; [email protected] (T.M.); [email protected] (K.L.); [email protected] (X.Y.);
[email protected] (J.Y.); [email protected] (S.W.)
2 Key Laboratory of Food Nutrition and Safety, Ministry of Education of China,
Tianjin University of Science and Technology, Tianjin 300457, China
* Correspondence: [email protected]; Tel.: +86-022-6091-2493

Abstract: In this study, a zearalenone (ZEN) hapten was designed and prepared against the mycotoxin
ZEN, and the original coating ZEN-ovalbumin (ZEN-OVA) was prepared by conjugation with OVA.
Based on the gold nanorods (AuNRs) of uniform size and stable properties synthesized by the seed-
mediated method, the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and the
AuNRs growth-based multicolor ELISA for detecting ZEN toxin were further established. Under
the optimal experimental conditions, the coating amount of ZEN-OVA: 0.025 μg/well, antibody
(Ab) dilution factor: 32,000 times, blocking solution: 0.5% skimmed milk powder, enzyme-labeled
secondary Ab diluted 10,000 times, and a pH of the PBS buffer at 7.4, the sensitivity (IC50 ) of the
established ic-ELISA for ZEN detection reached 0.85 ± 0.04 μg/L, and the limit of detection (IC15 )
Citation: Ma, T.; Liu, K.; Yang, X.;
Yang, J.; Pan, M.; Wang, S.
reached 0.22 ± 0.08 μg/L. In the multicolor ELISA based on the growth of AuNRs, as the content
Development of Indirect Competitive of ZEN increased, the mixed solution exhibited a significant color change from brownish red to
ELISA and Visualized Multicolor colorless. ZEN concentration as low as 0.1 μg/L can be detected by the naked eye (brown red to
ELISA Based on Gold Nanorods dark gray). This study provided an effective analysis strategy for the rapid screening and accurate
Growth for the Determination of monitoring of the ZEN contaminant in foods.
Zearalenone. Foods 2021, 10, 2654.
https://doi.org/10.3390/ Keywords: zearalenone; gold nanorods; indirect competitive ELISA; visualized multicolor ELISA
foods10112654

Academic Editor: Andrew G. Gehring

1. Introduction
Received: 6 October 2021
Accepted: 28 October 2021
Zearalenone (ZEN) is a nonsterol estrogen mycotoxin mainly produced by Fusarium
Published: 2 November 2021
genera, which is widely found in grain crops, such as corn, wheat, and barley [1,2]. The
degree of contamination of food crops by Fusarium mainly depends on the moisture and
Publisher’s Note: MDPI stays neutral
temperature conditions; the optimal growth temperature for Fusarium is between 24 and
with regard to jurisdictional claims in
32 ◦ C and the optimal humidity is 40% [3]. Therefore, in areas with sufficient rainfall and
published maps and institutional affil- high relative humidity, cereals are likely to be contaminated with Fusarium in all steps of
iations. production, storage, and processing [4]. ZEN can cause excessive estrogen syndrome in
animals such as pig, poultry, and humans, as well as immunotoxicity, genotoxicity, and
suspected carcinogenicity [5,6]. It has been reported that the potential etiological mecha-
nism of breast cancer involves changes in the cytochrome P450 (CYP) enzyme, which is
Copyright: © 2021 by the authors.
related to ZEN [7]. Yu et al. have demonstrated that the ZEN on 2,3,7,8-Tetrachlorodibenzo-
Licensee MDPI, Basel, Switzerland.
p-dioxin (TCDD)-induced CYP1A1 activity and gene expression involved the estrogen
This article is an open access article
receptor pathway [8]. In addition, due to its similar structure to endogenous estrogen,
distributed under the terms and ZEN can show estrogen activity in vivo and competitively bind with estrogen receptors,
conditions of the Creative Commons thus affecting estrogen secretion in humans or animals, resulting in reproductive organ
Attribution (CC BY) license (https:// abnormalities, infertility, abortion, and other diseases [9–11]. At present, many countries
creativecommons.org/licenses/by/ and organizations have regulated the maximum residue levels (MRLs) of ZEN in foods,
4.0/). although a consistent standard has not yet been obtained (European Union: 20–400 μg/kg

Foods 2021, 10, 2654. https://doi.org/10.3390/foods10112654 https://www.mdpi.com/journal/foods

6
Foods 2021, 10, 2654

in different types of foods, Russia: 1000 μg/kg in hard wheat, flour and wheat germ; China:
60 μg/kg in cereals and its products) [12–14]. Therefore, the development of effective
strategies for ZEN detection in foods is of great significance for protecting the health of
humans and animals.
Currently, traditional instrumental methods based on liquid chromatography (LC),
gas chromatography (GC), and mass spectrometry (MS) have been widely applied in the
detection of ZEN contaminant in foods and animal feed samples [15–20]. However, these
techniques require sophisticated instruments, professional and experienced operators, and
long test time, which limits the practical application to a certain extent. Engvall and Perl-
mann achieved the quantitative detection of a solid phase enzyme immunoassay for the
first time, marking the successful construction of an enzyme linked immunosorbent assay
(ELISA) [21,22]. The ELISA has the characteristics of simple operation, high specificity,
and low cost, and has gradually become the mainstream technology in the field of rapid
detection [23,24]. Improving the sensitivity and lowering the detection limit of ELISA can
promote the further application of this effective technology [25,26]. Signal amplification has
been demonstrated by many researchers to improve the sensitivity of ELISA. In particular,
the introduction of various nanomaterials has further improved the performance of tradi-
tional ELISA. For example, Zhang et al. synthesized CdTe/CdS/ZnS quantum dots (QDs)
in the aqueous phase and developed a fluorescent immunoassay (FLISA) for detecting ZEN
in corn [27]. Xiong et al. reported an advanced enzyme-assisted etching method in which
gold nanorods (AuNRs) were applied as the signal carrier for aflatoxin B1 (AFB1 ) in corn
samples to amplify the ELISA signal [28]. Liu et al. constructed a horseradish peroxidase
(HRP)-mediated ratio fluorescence ELISA based on gold and silver bimetallic nanoclusters
(Au-AgNCs) to detect zearalenone, which significantly improved the detection limit [29].
In this study, we successfully prepared the ZEN hapten and ZEN coating antigen
(ZEN-ovalbumin (ZEN-OVA)) and developed an indirect competitive-ELISA (ic-ELISA)
strategy using monoclonal antibodies (anti-ZEN-Abs) with high sensitivity and specificity.
On this basis, a visual multicolor ELISA was developed based on alkaline phosphatase
(AP) converting ascorbic acid–phosphate (VcP) to ascorbic acid (Vc) to control the growth
of AuNRs. Compared with traditional ic-ELISA for ZEN, the visualized multicolor ELISA
based on AuNRs growth offered a more convenient and intuitive strategy for the detection
of ZEN contaminant (Scheme 1).

Scheme 1. Procedure of the visualized AuNRs growth-based multicolor ELISA for ZEN detection
compared with the traditional ic-ELISA.

7
Foods 2021, 10, 2654

2. Materials and Methods

2.1. Material and Apparatus
Hexadecyl trimethyl ammonium bromide (CTAB, 99%) was purchased from Solarbio
(Beijing, China). Chloroauric acid (HAuCl4 ) for the synthesis of AuNRs was obtained
from Sigma-Aldrich (St. Louis, MO, USA). Silver nitrate (AgNO3 ), Vc, VcP, and sodium
borohydride (NaBH4 ) for the synthesis of AuNRs, 1-ethyl-3-[3-(dimethylamino) propyl]
carbodimide (EDC) and OVA for the synthesis of the conjugate of ZEN-OVA were obtained
from the Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Pyridine and car-
boxymethoxylamine (CMO) were purchased from TCI Development Co. Ltd. (Shanghai,
China). The anti-ZEN monoclonal antibody (anti-ZEN-Ab, 1.0 mg/mL) was purchased
from Shandong Lvdu Biotechnology Co. Ltd. (Shandong, China). 3,3,5,5-tetramethylbenzi-
dine (TMB), HRP goat anti-mouse IgG conjugated (1.0 mg/mL), AP goat anti-mouse
IgG conjugated (3.5 mg/mL), AFB1 , ZEN and the structural analogues (α-zearalenol, α-
zearalanol, β-zearalenol, zearalanone, β-zearalanol) (1.0 mg/L) were also purchased from
Sigma-Aldrich (St. Louis, MO, USA). T-2 toxin, ochratoxin A (OTA), and fumonisin B2
(FB2 ) were purchased from Toronto Research Chemicals (Toronto, ON, Canada).
The microplate reader for reading the absorbance values was purchased from Thermo
Fisher Scientific (Waltham, MA, USA). The UV-visible spectrophotometer (Cary 50 Bio)
was obtained from Varian (Salt Lake, CA, USA). The 96-well polystyrene microplates, mul-
tichannel pipettes (100–300 μL), and single-channel pipettes (2.5–1000 μL) were obtained
from Thermo Fisher Scientific (Waltham, MA, USA). The transmission electron microscopy
(TEM) images were obtained from Talos G2 200X electronic microscope (Thermo Fisher
Scientific (Waltham, MA, USA). A vortex machine (HQ-60) was purchased from North
Tongzheng Biotechnology Development Company (Beijing, China). Milli-Q Ultrapure
Water System was purchased from Milli-Q Millipore, (Bedford, MA, USA).

2.2. Preparation of the Coating Antigen—The Conjugate of ZEN-OVA

The preparation method of ZEN-CMO was modified according to the previous liter-
ature [30]. Briefly, 10.0 mg of ZEN was dissolved in 200 μL of methanol and mixed well.
Then, 20.0 mg of CMO and 1.0 mL of anhydrous pyridine were sequentially added into
the above mixed solution. The mixture was stirred and reacted under nitrogen protection
for 24 h. After the reaction was completed, the mixture was placed in a vacuum oven at
80 ◦ C to remove pyridine and store at 4 ◦ C for later use. We mixed 9.78 mg of the above
conjugate ZEN-CMO and 14.0 mg of EDC, successively dissolved it in 1.0 mL of DMF, and
stirred and activated it overnight at 4 ◦ C. In total, 10.0 mg of OVA was dissolved in 2.0 mL
of NaHCO3 (130 mmol/L) and precooled. The activated product was added dropwise
under ice bath conditions. Two hours later, the product was placed under 4 ◦ C overnight
and dialyzed against PBS buffer solution for 72 h and stored at −20 ◦ C.

2.3. Test Procedure of ic-ELISA

The antigen ZEN-OVA was dissolved in the coating solution and mixed evenly. The
mixture was added to a microplate (0.025, 0.05, 0.1 μg/well, 100 μL/well), and incubated
overnight at 4 ◦ C. After washing the plate with PBST (0.01 mol/L PBS and 0.1% Tween-20)
3 times, the blocking solution (PBS containing 0.5% skimmed milk powder, 200 μL/well)
was added, and we incubated the mixture at 37 ◦ C for 1 h. After washing, the mixture of
ZEN standards were diluted to different concentrations (50 μL) and anti-ZEN Ab (50 μL)
was added and reacted at 37 ◦ C for 1 h. Then, 100 μL of HRP goat anti-mouse IgG was
added into each well and incubated for 0.5 h. After washing the plate with PBST 5 times,
100 μL of the TMB substrate solution was added into each well. After incubation for
15–30 min, the reaction was terminated with 50 μL of H2 SO4 (1.25 mol/L). Then, the
absorbance was measured at 450 nm using a microplate reader.

8
Foods 2021, 10, 2654

2.4. Specificity of ic-ELISA

The specificity of Abs is related to the structure of the antigenic determinant and
is expressed as the cross-reactivity rate. Ten mycotoxins including ZEN, five structural
analogs of ZEN (α-zearalenol, β-zearalenol, α-zearalanol, β-zearalanol, and zearalanone),
and four common mycotoxins (AFB1 , OTA, T-2 toxin, and FB2 ) were used as competing
standards to determine the cross-reactivity rate to anti-ZEN-Ab by ic-ELISA. The mentioned
analytes were diluted with PBS to the following concentrations (1000, 333.3, 111.11, 37.03,
12.34, 4.11, 1.37, 0.45, 0.15, 0.05, and 0.016 μg/L). The following equation was used to
calculate the cross-reaction rate.

CR (%) = IC50 (50% inhibitory concentration (ZEN))/IC50 (50% inhibitory

(1)
concentration (competitor)) × 100%

2.5. AuNR Growth-Based Multicolor ELISA for ZEN

Preparation of AuNRs seed liquid. The AuNRs were synthesized by the seed-
mediated method in this study. A total of 1.0 mL of CTAB (0.2 mol/L) and 1.0 mL of
HAuCl4 (0.5 mmol/L) were added into a round-bottom flask and mixed thoroughly. Then,
120 μL of NaBH4 (0.01 mol/L) was added and stirred gently for 2 min. The obtained
mixture was used as the seed liquid for AuNR growth.
Effect of Vc dosage on AuNR growth. The mixed solution containing 125 μL of
CTAB (0.2 mol/L), 1.5 μL of AgNO3 (0.01 mol/L), and 12.5 μL of HAuCl4 (0.01 mol/L) was
added to each well of the enzyme-labeled plate. After mixing, the Vc (0.01 mol/L) solution
of different volumes (0–20 μL) was added to control the total volume of the solution to
240 μL. Next, 10 μL of the seed solution was added to mediate the production of AuNRs.
After mixing evenly, the mixture was incubated at room temperature for 1 h, and UV
absorption spectra of the grown AuNRs were recorded.
Procedure of visualized multicolor ELISA based on AuNR growth. The encapsula-
tion process of the ZEN-OVA conjugate and its competitive binding with anti-ZEN Abs
were the same as ic-ELISA. The difference was that 100 μL of AP-goat anti-mouse IgG
diluted 10,000 times with Tris-HCl buffer (1 mmol/L pH 7.4) was added to each well. After
incubation at 37 ◦ C for 30 min and full washing, 80 μL of VcP (15 mmol/L) was added
into each well. After full mixing, the mixture was incubated at 37 ◦ C for 1 h. Then, 50 μL
of the above reaction solution was mixed evenly with the solution containing 125 μL of
CTAB (0.2 mol/L), 1.5 μL of AgNO3 (0.01 mol/L), and 12.5 μL of HAuCl4 (0.01 mol/L)
and controlled to 240 μL with ultrapure water. After 10 μL of seed solution was added,
the mixture was incubated at room temperature for 1 h and tested by naked eye and UV
absorption spectroscopy.

3. Results and Discussion

3.1. Preparation and Characterization of ZEN-CMO and ZEN-OVA Conjugate
In this study, the prepared ZEN-CMO (Mr: 391.28) was analyzed by mass spectrom-
etry (Figure S1). In an anionic environment, the characteristic ion peaks of ZEN-CMO
[M-1]¯ at 390.40 and [2M-1]− at 781.80 were observed, proving the successful preparation
of the hapten ZEN-CMO with acceptable purity. The ZEN-CMO product was further
conjugated with OVA to obtain ZEN-OVA conjugate, the concentration of which was deter-
mined to be 3.85 mg/mL by a commercial BCA kit. Furthermore, the prepared ZEN-OVA
conjugate was tested for its conjugate ability to anti-ZEN Abs (Table S1). When the Abs was
diluted 16,000 times, the OD450 value reached 1.253, and the inhibition rate reached 98.92%,
indicating the successful coupling of ZEN-CMO and OVA, and the obtained ZEN-OVA
conjugate could be used for subsequent immunoassay experiments.

3.2. Conditions Optimization of Traditional ELISA

Dilution times of ZEN-OVA conjugate and anti-ZEN Abs. Different amounts of
ZEN-OVA conjugate were coated on a 96-well microtiter plate to optimize the dilution

9
Foods 2021, 10, 2654

times through ic-ELISA. According to the results shown in Table S2, when the coating
amount of the ZEN-OVA conjugate was 0.1 μg/well and the Ab dilution factor was 64,000,
the OD450 value reached 0.858, and the IC50 was 2.68 μg/L. When the coating amount was
0.05 μg/well and 0.025 μg/well, the dilution factor was 32,000, the corresponding OD450
and IC50 values were 1.078 and 0.793, 1.06 μg/L and 0.85 μg/L, respectively. It can be
clearly seen that the IC50 increased with the increase in the coating amount of ZEN-OVA,
which was because the excessive amount of ZEN-OVA bound to the Ab led to the decrease
in detection sensitivity. When the coating amount was 0.025 μg/well and the Ab dilution
factor was 32,000, the color development was relatively stable and the IC50 reached the
lowest value, which was selected as the optimal condition for subsequent experiments.
Blocking Solution. The blocking solution was applied to block the excess binding
sites in the micropores. A higher concentration of the blocking solution may affect the
subsequent binding between the antigen and Ab, thereby reducing the sensitivity of the
method. In the study, the IC50 values of the ic-ELISA method using different concentrations
(0.5% and 1%) of skimmed milk powder as the blocking solution were compared (Table S3).
The 0.5% skimmed milk powder had an IC50 of 0.85 μg/L, less than using 1% skimmed
milk powder (2.41 μg/L), which was selected as the blocking solution.
pH value of PBS diluent. PBS buffers with different pH values (5.7, 7.4, and 8.5) were
used for Ab dilution and ic-ELISA tests were performed (Table S4). When PBS was used at
pH 7.4, the IC50 value was only 0.85 μg/L, which was significantly lower than that obtained
at pH 5.7 and 8.5 (2.95 μg/L and 1.86 μg/L). At this time, the corresponding absorbance
value (λ = 450 nm) was also significantly higher than the other two pH values, because
the acidic or alkaline environment affected the binding reaction between the antigen and
Ab as well as enzyme-labeled antibody, resulting in the reduction in detection sensitivity.
Therefore, a PBS buffer of pH 7.4 was chosen as a diluent for Ab and ZEN standards.

3.3. ic-ELISA Standard Curve

Under the optimal conditions: coating amount of ZEN-OVA 0.025 μg/well, Ab dilu-
tion 32,000 times, blocking solution 0.5% skim milk powder in PBS, PBS buffer (pH 7.4) as
diluent, the standard curve of ic-ELISA for ZEN is shown in Figure 1. The sensitivity (IC50 )
and limit of detection (IC15 ) reached 0.85 ± 0.04 μg/L and 0.22 ± 0.08 μg/L, indicating
that this method can provide accurate and sensitive analysis for the ZEN toxin.

Figure 1. Standard curve of ZEN by ic-ELISA.

3.4. Specificity of Traditional ELISA

In order to evaluate the specificity of the established ic-ELISA method to ZEN, five
ZEN structural analogs (α-zearalanol, β-zearalanol, α-zearalenol, β-zearalenol, and zear-

10
Foods 2021, 10, 2654

alenone) and four common mycotoxins (AFB1 , OTA, FB2 , and T-2 toxin) were selected
and analyzed (Table 1). The cross-reaction rate of four structural analogs α-zearalanol,
β-zearalanol, α-zearalenol, and β-zearalenol were 35.27%, 45.70%, 29.72% and 17.93%,
respectively; the cross-reaction rate of zearalenone was only 1.58%; and there was no
obvious cross-reaction with other mycotoxins. These results proved that the established
ic-ELISA method had high specificity.

Table 1. Cross-reacting of ZEN with other mycotoxins.

Determinand Structure IC50 (μg/L) Cross-Reaction Rate

Zearalenone 0.85 100

α-Zearalanol 2.41 35.27

β-Zearalanol 1.86 45.70

α-Zearalenol 2.86 29.72

β-Zearalenol 4.74 17.93

Zearalanone 53.79 1.58

AFB1 >1000 <0.01

&O

+ 2
2 1
OTA 2+ >1000 <0.01
2 2+ 2

FB2 >1000 <0.01

+ + 2+
+ & 2
&+ 2 
2 2
T-2 toxin +& 2 >1000 <0.01
2 2 &+
&+
+& 2

11
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- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Part 5: Statistical analysis and interpretation
Note: Key terms and definitions
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Critical analysis and evaluation
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 42: Literature review and discussion
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Remember: Assessment criteria and rubrics
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 44: Diagram/Chart/Graph]
Key Concept: Best practices and recommendations
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Research findings and conclusions
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Remember: Fundamental concepts and principles
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
[Figure 47: Diagram/Chart/Graph]
Practice Problem 47: Historical development and evolution
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Note: Problem-solving strategies and techniques
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Critical analysis and evaluation
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Introduction 6: Best practices and recommendations
Note: Best practices and recommendations
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Interdisciplinary approaches
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
[Figure 52: Diagram/Chart/Graph]
Note: Practical applications and examples
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 53: Diagram/Chart/Graph]
Definition: Comparative analysis and synthesis
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Learning outcomes and objectives
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Assessment criteria and rubrics
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 56: Problem-solving strategies and techniques
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Practical applications and examples
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Theoretical framework and methodology
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Theoretical framework and methodology
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Conclusion 7: Literature review and discussion
Note: Ethical considerations and implications
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Remember: Practical applications and examples
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 62: Theoretical framework and methodology
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 63: Key terms and definitions
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Literature review and discussion
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 65: Problem-solving strategies and techniques
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Study tips and learning strategies
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 67: Diagram/Chart/Graph]
Key Concept: Best practices and recommendations
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 68: Diagram/Chart/Graph]
Important: Case studies and real-world applications
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Theoretical framework and methodology
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Abstract 8: Literature review and discussion
Definition: Theoretical framework and methodology
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 71: Diagram/Chart/Graph]
Remember: Ethical considerations and implications
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Current trends and future directions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Theoretical framework and methodology
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 74: Assessment criteria and rubrics
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Best practices and recommendations
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Remember: Study tips and learning strategies
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 77: Experimental procedures and results
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Fundamental concepts and principles
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Historical development and evolution
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Introduction 9: Case studies and real-world applications
Example 80: Literature review and discussion
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 81: Diagram/Chart/Graph]
Practice Problem 81: Assessment criteria and rubrics
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Comparative analysis and synthesis
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
[Figure 83: Diagram/Chart/Graph]
Remember: Historical development and evolution
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Interdisciplinary approaches
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 85: Diagram/Chart/Graph]
Example 85: Historical development and evolution
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Fundamental concepts and principles
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Assessment criteria and rubrics
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 88: Key terms and definitions
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Learning outcomes and objectives
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
References 10: Assessment criteria and rubrics
Important: Statistical analysis and interpretation
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Experimental procedures and results
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Note: Statistical analysis and interpretation
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Interdisciplinary approaches
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Key Concept: Current trends and future directions
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Practice Problem 95: Current trends and future directions
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Study tips and learning strategies
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Key Concept: Comparative analysis and synthesis
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Note: Research findings and conclusions
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Key Concept: Historical development and evolution
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 100: Diagram/Chart/Graph]
Results 11: Literature review and discussion
Practice Problem 100: Historical development and evolution
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 101: Diagram/Chart/Graph]
Key Concept: Statistical analysis and interpretation
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Ethical considerations and implications
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 103: Critical analysis and evaluation
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Case studies and real-world applications
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Ethical considerations and implications
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Important: Literature review and discussion
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 107: Theoretical framework and methodology
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Problem-solving strategies and techniques
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 109: Diagram/Chart/Graph]
Definition: Best practices and recommendations
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 110: Diagram/Chart/Graph]
Introduction 12: Best practices and recommendations
Practice Problem 110: Study tips and learning strategies
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Historical development and evolution
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Example 112: Theoretical framework and methodology
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 113: Diagram/Chart/Graph]
Important: Statistical analysis and interpretation
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
[Figure 114: Diagram/Chart/Graph]
Definition: Best practices and recommendations
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Comparative analysis and synthesis
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Learning outcomes and objectives
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 117: Diagram/Chart/Graph]
Remember: Study tips and learning strategies
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 118: Interdisciplinary approaches
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 119: Learning outcomes and objectives
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Appendix 13: Fundamental concepts and principles
Definition: Learning outcomes and objectives
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Assessment criteria and rubrics
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Important: Learning outcomes and objectives
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Experimental procedures and results
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 124: Research findings and conclusions
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Statistical analysis and interpretation
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Key Concept: Case studies and real-world applications
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Note: Practical applications and examples
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Key terms and definitions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Example 129: Experimental procedures and results
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 130: Diagram/Chart/Graph]
Topic 14: Practical applications and examples
Note: Experimental procedures and results
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Experimental procedures and results
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Remember: Statistical analysis and interpretation
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 133: Diagram/Chart/Graph]
Key Concept: Key terms and definitions
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Interdisciplinary approaches
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Definition: Problem-solving strategies and techniques
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Theoretical framework and methodology
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Best practices and recommendations
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Learning outcomes and objectives
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 139: Statistical analysis and interpretation
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
References 15: Literature review and discussion
Example 140: Critical analysis and evaluation
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 141: Diagram/Chart/Graph]
Example 141: Assessment criteria and rubrics
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 142: Fundamental concepts and principles
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Literature review and discussion
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Problem-solving strategies and techniques
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Definition: Comparative analysis and synthesis
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 146: Diagram/Chart/Graph]
Key Concept: Learning outcomes and objectives
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Note: Theoretical framework and methodology
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Interdisciplinary approaches
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 149: Diagram/Chart/Graph]
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