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FOOD CHEMISTRY · 2023 · IF 8.8

On-site quantitation of xanthine in fish and serum using a smartphone-based spectrophotometer integrated with a dual-readout nanosensing assay

Ningxia Xu,Meng Xiao,Zipei Yu,Baohui Jin,Mengsu Yang,Changqing Yi

Rapid and quantitative biochemical analysis at points-of-need is imperative for food safety inspection. This work reports on: 1) a stand-alone smartphone-based “two-in-one” spectrophotometer (the SAFS) installed with a self-developed application (the SAFS-App) which can precisely collect both absorption spectra and fluorescence spectra in a reproducible manner within 5 s; and 2) a straightforward protocol for xanthine detection using fluorescent carbon nanodots and silver nanoparticles. The assay performed with the SAFS demonstrates high specificity towards xanthine, and a linear range of 1–60 μM with LODs of 0.38 and 0.58 μM for colorimetric and fluorometric readouts, respectively. The reliability and robustness of the SAFS are validated by on-site quantitation of xanthine in fish and serum samples, with comparable accuracy to HPLC method. More importantly, the SAFS presents itself as an appealing device which is accessible to everyone through the Internet of Things and can be tailored for diverse point-of-care testing applications.

研究领域: 分析化学 · 食品科学 · 生物医学工程
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ANALYTICAL CHEMISTRY · 2016 · IF 5.886

Fluorescent and Colorimetric Dual-Readout Assay for Inorganic Pyrophosphatase with Cu2+-Triggered Oxidation of o-Phenylenediamine

Jian Sun,Bin Wang,Xue Zhao,Zong-Jun Li,Xiurong Yang

We demonstrate a rationally designed fluorescent and colorimetric dual-readout strategy for the highly sensitive, quantitative determination of inorganic pyrophosphatase (PPase) activity, a key hydrolytic enzyme involved in a variety of metabolic processes. Inspired by the selective oxidative and chromogenic reaction of o-phenylenediamine (OPD) with Cu2+, the special inhibitory effects of pyrophosphate (PPi) on the oxidative ability of Cu2+, and the specific hydrolysis of PPi into orthophosphate by PPase, a convenient small molecule OPD-based analytical system was developed for Cu2+/PPi recognition and PPase activity assay. We have confirmed that Cu2+ acts as the oxidant in the reaction and the main chromogenic product of OPD is 2,3-diaminophenazine (usually called OPDox) in the assay by combining the ESI-MS, 1H NMR, and XPS spectra analysis. Direct electrochemical insights into the Cu2+-triggered and PPi-inhibited mechanism were performed by cyclic voltammetry characterizations of the Cu2+ in the absence and presence of PPi for the first time. Furthermore, the proposed analytical system with clear response mechanism exhibits a promising outlook for the PPase activity assay in real biological samples and inhibitor screening.

研究领域: 生物化学 · 分析化学
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ACS Sustainable Chemistry & Engineering · 2023 · IF 8.4

Multifunctional Probe Based on CTAB-Cu Nanoparticles for Fluorescence and Colorimetric Dual-Read-Out Determination of p-Nitrophenol and Glyphosate

Huiwen Wang,Sihao Wu,Zoujun Peng,Xiulin Wang,Juanhua Lai,Ping Qiu

The quantitative analysis of glyphosate has significant implications for the evaluation of food quality and environmental safety risks. First, copper nanoparticles (Cu NPs) were synthesized using cetyltrimethylammonium bromide (CTAB) as a stabilizing agent, which resolved the limitation associated with atmospheric oxidation of Cu NPs and provided excellent fluorescence stability. Interestingly, the fluorescence of blue-emitting CTAB-Cu NPs was quenched by pH-responsive p-nitrophenol (p-NP) via Förster resonance energy transfer. Herein, a dual read-out sensor based on CTAB-Cu NPs and p-NP was constructed for glyphosate detection. Specifically, acetic acid (HAc) from acetylcholine (ACh) via hydrolysis catalyzed by acetyl cholinesterase (AChE) reduced the pH of the mixed solution and prevented the loss of protons from p-NP, which allowed the fluorescence emission by CTAB-Cu NPs. The presence of glyphosate inhibited the activity of AChE, and the increased pH resulted in proton loss. The fluorescence intensity of CTAB-Cu NPs was quenched, and the solution color changed from colorless to yellow with a UV absorption peak at 400 nm. The sensor can detect ultra-trace levels of glyphosate residues with a limit of detection of 5.43 ng/L. The obtained biosensor was successfully utilized to monitor the degradation of glyphosate in the roots of cabbage and cultivated soil.

CAS / MDL: 100-02-7
研究领域: 分析化学 · 环境科学 · 食品科学
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RSC Advances · 2024 ·Vol. 14 ·期 11 ·7858-7866 · IF 3.9

Highly sensitive and accurate detection of cholesterol based on a single red upconversion biosensor

Pang Tingyuan,Liu Xiaorui,Li Jia,Song Qi,Li Junren,Han Ling,Shu Wenying,Jian Xiaoshun,Zhang Meimei

Cholesterol (CHOL) is an important clinical biochemical indicator that plays an important role in the regulation of the fluidity, permeability, and microstructure of cell membranes. Therefore, it is necessary to accurately monitor CHOL levels in biological samples for the early prevention and diagnosis of various diseases. The single-band red upconversion nanoparticle (UCNP) emits light within the optical transmission window of biological tissues, and can penetrate deeper biological tissues and cause less energy loss due to scattering and thus have higher sensitivity and accuracy. Here, using the nontoxic, sensitive, and photochemically stable 3,3′,5,5′-tetramethylbenzidine (TMB) as the quenching agent and single red UCNP as the fluorescent donor, a dual-readout colorimetric and fluorescent sensor was developed to detect CHOL. The detection mechanism and feasibility were discussed in detail, and experimental conditions such as Fe2+ concentration, TMB concentration and reaction time were explored. Under optimal conditions, the limits of CHOL detection by colorimetry and fluorescence were 0.85 μM and 0.63 μM. The sensing system was used to measure CHOL in serum samples and the values obtained by these two modes were close, and the spiked recoveries were 97.2–102.2% and 97.1–103.7%, respectively, which holds great potential in clinical diagnosis and health management.

研究领域: 生物化学 · 医学诊断 · 纳米技术
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ANALYTICAL CHEMISTRY · 2019 · IF 6.35

Dual-Readout Tyrosinase Activity Assay Facilitated by a Chromo-Fluorogenic Reaction between Catechols and Naphthoresorcin

Jiahui Zhao,Guoyong Liu,Jian Sun,Qifeng Wang,Zong-Jun Li,Xiurong Yang

Analyte-responsive chromo-fluorogenic reactions under accessible conditions are important for designing small-molecule spectroscopic probes. We describe a series of newly constructed motifs based on the chromo-fluorogenic reaction between catechol derivatives (typically hydroxytyrosol, dopamine, and levodopa) and naphthoresorcin (NR) in aqueous solution under ambient conditions. The weakly absorptive and fluorogenic catechols/NR was converted to products having visible absorption and bright fluorescence within several minutes. The chromo-fluorophores produced from this reaction had a maximum absorbance at 458 nm and emission at 480 nm with high fluorescence quantum yields (30–84%). Inspired by the tyrosinase-catalyzed hydroxylation of monophenols to catechols, the tyrosinase-enabled chromo-fluorogenic reaction was verified by using monophenol (typically tyrosol) as the substrate. In this regard, a dual-readout tyrosinase activity assay was developed by virtue of the in situ “turn-on” optical signals. Furthermore, a test of tyrosinase inhibition, by using a common inhibitor kojic acid, demonstrated the potential of the chromo-fluorogenic reaction for developing other tyrosinase related assays and signal transduction.

CAS / MDL: 2206-27-1 7722-84-1
研究领域: 生物化学 · 分析化学 · 医学诊断
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Journal of Materials Chemistry B · 2018 ·Vol. 7 ·期 2 ·233-239 · IF 4.776

Pd nanoparticle-decorated graphitic C3N4 nanosheets with bifunctional peroxidase mimicking and ON–OFF fluorescence enable naked-eye and fluorescent dual-readout sensing of glucose

Wenchi Zhang,Xin Li,Xuechao Xu,Yanfang He,Fengxian Qiu,Jianming Pan,Xiangheng Niu

In this work, we report a novel naked-eye and fluorescent dual-readout glucose sensing platform based on the bifunctional peroxidase mimicking and ON–OFF fluorescence of Pd nanoparticle-decorated graphitic C3N4 nanosheets (PdNPs/g-C3N4). The PdNPs/g-C3N4 hybrid not only exhibited a strong peroxidase-like activity to catalyze colorless o-phenylenediamine (OPD) into its yellow product OPDox in the presence of H2O2, but it also acted as a turn-off fluorescent sensor for the assembly of the generated OPDox on g-C3N4. When coupled with glucose oxidase (GOx), the platform could provide both visual and fluorescent responses for glucose specifically. Under optimal conditions, the platform was able to detect the target at concentrations as low as 50 μM and 0.4 μM in the linear ranges of 50–2000 μM and 1–1000 μM with the naked eye and a fluorometer, respectively. The developed platform integrates the equipment-free merit of naked-eye observation with the excellent performance of fluorometric analysis well, providing a new and powerful tool for both rough assessment and accurate detection of glucose in diabetes management and other applications.

CAS / MDL: 7647-10-1
研究领域: 分析化学 · 生物医学工程 · 材料科学
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FOOD CHEMISTRY · 2024 · IF 8.5

Novel litchi-like Au–Ag nanospheres driven dual-readout lateral flow immunoassay for sensitive detection of pyrimethanil

Xinhui Shu,Ping Guo,Gan Zhang,Wei Zhang,Hong Hu,Juan Peng,Yonghua Xiong,Bingfeng Ma,Weihua Lai

Pyrimethanil (PYR) is a fungicide that is harmful to consumers when present in foods at concentrations greater than maximum permitted residue levels. High-performance immunoprobes and dual-readout strategy may be useful for constructing sensitive lateral flow immunoassay (LFIA). Herein, the prepared litchi-like Au–Ag bimetallic nanospheres (LBNPs) exhibited high mass extinction coefficients and fluorescence quenching constants. Benefiting from LBNPs and dual-readout mode, the limits of detection of LBNPs-CM-LFIA and LBNPs-FQ-LFIA for PYR were 0.957 and 0.713 ng mL −1 , which were 2.54- and 3.41-fold lower than that of gold nanoparticles-based LFIA, respectively. The limits of quantitation of LBNPs-CM-LFIA and LBNPs-FQ-LFIA were 3.740 and 1.672 ng mL −1 , respectively. LBNPs-LFIA was applied to detect PYR in cucumber and grape samples with satisfactory recovery (90%–111%). LBNPs-LFIA showed good agreement with LC-MS/MS for the detection of PYR in the samples. Accordingly, this sensitive and accurate dual-readout LFIA based on LBNPs can be effectively applied for food safety.

研究领域: 食品安全检测 · 纳米材料 · 分析化学
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JOURNAL OF HAZARDOUS MATERIALS · 2024 · IF 12.2

Self-assembled bifunctional nanoflower-enabled CRISPR/Cas biosensing platform for dual-readout detection of Salmonella enterica

Manyan Qiu,Zhiyu Yuan,Nan Li,Xinyan Yang,Xianlong Zhang,Yujun Jiang,Qianyu Zhao,Chaoxin Man

Sensitive detection and point-of-care test of bacterial pathogens is of great significance in safeguarding the public health worldwide. Inspired by the characteristics of horseradish peroxidase (HRP), we synthesized a hybrid nanoflower with peroxidase-like activity via a three-component self-assembled strategy. Interestingly, the prepared nanozyme not only could act as an alternative to HRP for colorimetric biosensing, but also function as a unique signal probe that could be recognized by a pregnancy test strip. By combining the bifunctional properties of hybrid nanoflower, isothermal amplification of LAMP, and the specific recognition and non-specific cleavage properties of CRISPR/Cas12a system, the dual-readout CRISPR/Cas12a biosensor was developed for sensitive and rapid detection of Salmonella enterica . Moreover, this platform in the detection of Salmonella enterica had limits of detection of 1 cfu/mL (colorimetric assay) in the linear range of 10 1 –10 8 cfu/mL and 10 2 cfu/mL (lateral flow assay) in the linear range of 10 2 –10 8 cfu/mL, respectively. Furthermore, the developed biosensor exhibited good recoveries in the spiked samples (lake water and milk) with varying concentrations of Salmonella enterica . This work provides new insights for the design of multifunctional nanozyme and the development of innovative dual-readout CRISPR/Cas system-based biosensing platform for the detection of pathogens. Environmental implication The presence of bacterial pathogens is pervasive in the environment and poses a significant threat to public health safety. To prevent foodborne disease outbreaks, it is crucial to develop sensitive and reliable pathogen detection strategies for rapid and on-site detection of pathogens in both environmental and food samples. Herein, we developed a novel dual-readout CRISPR/Cas biosensing platform based on nanozyme with bifunctional properties and loop-mediated isothermal amplification (LAMP) for highly sensitive and on-site detection of Salmonella enterica in lake water and milk samples. The proposed method has good stability and high accuracy in the detection of Salmonella enterica .

研究领域: 生物技术 · 纳米技术 · 病原体检测 · 生物传感器 · 即时诊断
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FOOD CHEMISTRY · 2025 · IF 9.8

Dual-readout lateral flow immunoassay for detection of amphenicols using intensified colorimetric–fluorescent carbon dots nanocomposite

Hongfang Li,Wenqing Xu,Yina Zhu,Hongxing Wei,Xiaoyue Liu,Lin Li,Guoliang Li

Conventional lateral flow immunoassays (LFIA) employing gold nanoparticles as probes suffer from insufficient sensitivity and poor accuracy in detecting amphenicols in food. Developing a dual-readout LFIA with fluorescence and colorimetric signals can significantly enhance its performance. Herein, fluorescent carbon dots were encapsulated into SiO 2 nanospheres to obtain the SiO 2 /CDs nanocomposites with a quantum yield of 81.26 %. The encapsulation enhances the signal intensity and prevents fluorescence quenching of carbon dots. Surface modification of SiO 2 /CDs with polydopamine (PDA) enables efficient antibody conjugation. The strong light absorption capacity of PDA also endowed the nanocomposites with good colorimetric signals. The SiO 2 /CDs@PDA-based LFIA showed 5-fold and 71-fold improvements in chloramphenicol detection using colorimetric and fluorescent modes, compared to conventional LFIA. The quantitative detection limits were 1.03–1.54 ng/g and 0.1–0.78 ng/g in colorimetric and fluorescent mode for amphenicols in chicken. The proposed LFIA offers a rapid, accurate, and reliable method for food safety monitoring.

CAS / MDL: 77-92-9 9048-46-8
研究领域: 免疫学 · 分析化学 · 纳米生物技术 · 食品安全 · 分子诊断 · 药物残留检测
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ANALYST · 2023 · IF 4.2

One-tube dual-readout biosensor for detection of nucleic acids and non-nucleic acids using CRISPR-ALP tandem assay

Xinxin Ke,Yi Hu,Chuanxia Chen,Tao Hu

Clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostics have been considered as the next generation molecular diagnosis tool. Single-readout mode has been extensively employed in massive CRISPR/Cas12a-based biosensors. In this work, we propose a one-tube dual-readout biosensor for ultrasensitive nucleic acids and non-nucleic acids detection by harnessing CRISPR-ALP tandem assay for the first time (termed CRISAT). In the presence of target, the Cas12a is activated to randomly cut the single-stranded hyDNA sequence of MB@hyDNA-cALP, thus releasing abundant alkaline phosphatase (ALP) in the supernatant solution. By using 4-aminophenol phosphate as the substrate of ALP, p-aminophenol is produced and then reacts with N-[3-(trimethoxysilyl)propyl]ethylenediamine or diethylenetriamine to generate silicon-containing polymer carbon dots (Si PCDs) or polymer carbon dots (PCDs) in situ, which can be observed by naked eye or detected by a fluorescent device in the same solution. Using this strategy, a fluoroscence and colorimetry dual-readout nanoplatform for CRISPR-based biosensor can be rationally developed. We ascertain the applicability of CRISAT through detecting SARS-CoV-2 pseudovirus, achieving superior sensitivity and specificity. With simple modification of crRNAs, the CRISAT platform can be also employed to detect monkeypox virus (MPXV) and non-nucleic acids of adenosine triphosphate (ATP). This work provides a great potential for nucleic acids and non-nucleic acids detection.

研究领域: 分子诊断 · 生物传感器 · 纳米技术 · 病毒学 · 分析化学
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BIOSENSORS & BIOELECTRONICS · 2021 · IF 10.618

An indicator displacement assay-based optical chemosensor for heparin with a dual-readout and a reversible molecular logic gate operation based on the pyranine/methyl viologen

Zipin Zhang,Rui Zhou,Yongliang Dong,Mengmeng Song,Lei Tian,Zuzhi Zhao

We have reported an optical indicator displacement assay (IDA) for heparin with a UV–vis absorbance and fluorescence dual-readout based on pyranine/methyl viologen (MV 2+ ). Upon introducing heparin, pyranine/MV 2+ shows a clearly observable increase in UV–vis absorbance and a turn-on of the fluorescence signal. We have demonstrated that the ionic nature of buffers significantly affects the pyranine displacement and the zwitterionic HEPES was most suitable for heparin sensing. After careful screening of experimental conditions, the pyranine/MV 2+ -based optical chemosensor exhibits a fast, sensitive, and selective response toward heparin. It shows dynamic linear concentration of heparin in the ranges of 0.1–40 U·mL −1 and 0.01–20 U·mL −1 for the absorptive and fluorescent measurements, respectively, which both cover the clinically relevant levels of heparin. As with the animal experiments, the optical chemosensor has been demonstrated to be selective and effective for heparin level qualification in rat plasma. The chemosensor is readily accessible, cost-effective, and reliable, which holds a great promise for potential application on clinical and biological studies. Furthermore, this IDA system can serve as an IMPLICATION logic gate with a reversible and switchable logical manner.

研究领域: 分析化学 · 生物化学 · 分子生物学 · 临床化学
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TALANTA · 2023 · IF 6.1

Simplifying the complexity: Single enzyme (choline oxidase) inhibition-based biosensor with dual-readout method for organophosphorus pesticide detection

Ziyu Yan,Zoujun Peng,Juanhua Lai,Peng Xu,Ping Qiu

Organophosphorus pesticides (OPs) are widely used in agricultural production, but their residues could cause pollution to the environment and living organisms. In this paper, a simple dual-readout method for OPs detection was proposed based on ChOx single enzyme inhibition. Firstly, ChOx can catalyze the production of H 2 O 2 from choline chloride (Ch-Cl). Bifunctional iron-doped carbon dots (Fe-CDs) with good peroxidase-like activity and superior fluorescence properties can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxidized TMB (oxTMB) by H 2 O 2 formed, and oxTMB could quench the fluorescence of Fe-CDs. In light of the fact that OPs exhibited activity in inhibiting ChOx, less H 2 O 2 and the decreasing oxTMB led to a result that the fluorescence of the system recovered and the solution became lighter in blue color. Moreover, the process of ChOx inhibition by OPs was analyzed by molecular docking technique and it was found that OPs interact with key amino acid residues catalyzed by ChOx (Asn510, His466, Ser101, His351, Phe357, Trp331, Glu312). Finally, a dual-mode (colorimetry and fluorescence) sensor was created for the detection of OPs with the detection limit of 6 ng/L, and was successfully used in the quantitative determination of OPs in actual samples with satisfactory results.

CAS / MDL: 298-00-0
研究领域: 农业化学 · 分析化学 · 环境科学
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