ANALYTICA CHIMICA ACTA
·
2024
·
IF 5.7
Chang Liu,Yuan Chen,Hong Lin,Limin Cao,Kaiqiang Wang,Xiudan Wang,Jianxin Sui
Background Due to the serious issue of ofloxacin (OFL) abuse, there is an increasingly urgent need for accurate and rapid detection of OFL. Immunoassay has become the “golden method” for detecting OFL in complex matrix beneficial to its applicability for a large-scale screening, rapidity, and simplicity. However, traditional antibodies used in immunoassay present challenges such as time-consuming preparation, unstable sensitivity and specificity, and difficulty in directional evolution. In this paper, we successfully developed an OFL detection method based on a shark-derived single-domain antibody (ssdAb) to address these issues. Results Using phage display technology and a heterologous expression system, OFL-specific clones 1O11, 1O13, 1O17, 1O19, 1O21, and 2O26 were successfully isolated and expressed in soluble form. Among all OFL-specific ssdAbs, the 1O17 ssdAb exhibited the highest binding affinity to OFL in a concentration-dependence manner. The limit of detection (IC 10 ) of 1O17 ssdAb was calculated as 0.34 ng/mL with a detection range of 3.40–1315.00 ng/mL, and its cross reactivity with other analogs was calculated to be less than 5.98 %, indicating high specificity and sensitivity. Molecular docking results revealed that 100Trp and 101Arg located in the CDR3 region of 1O17 ssdAb were crucial for OFL binding. In fish matrix performance tests, the 1O17 ssdAb did not demonstrate severe matrix interference in OFL-negative fish matrix, achieving satisfactory recovery rates ranging from 83.04 % to 108.82 % with high reproducibility. Significance This research provides a new and efficient OFL detection recognition element with significant potential in immunoassay applications, broadening the application scenarios of ssdAbs. It offers valuable insights into the structure-activity relationship between ssdAbs and small molecules, laying a theoretical foundation for the further directional modification and maturation of ssdAbs in subsequent applications.
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高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Di Zhao,Yanmei Shi,Chenqing Zhang,Zhihong Chen,Weixia Zhu,Shusheng Zhang,Kai Hu
A zwitterionic covalent organic framework was synthesized and applied as SPME blade coating for the extraction of fluoroquinolones in lake water and milk samples. The structure and functional groups of the sorbent were characterized via SEM, XPS, FT-IR, and XRD. Adsorption kinetic/isotherm analyses demonstrated chemisorption-driven monolayer adsorption with high affinity. Under the optimal SPME conditions, a high-throughput extraction method was established by incorporating zwitterionic covalent organic framework-coated blades into a 24-well plate system, which was subsequently coupled with LC-MS/MS for analysis. The proposed method exhibited good linearity (0.01–10 ng mL −1 , R 2 ≥ 0.9938), achieving LODs of 0.1–2.5 pg mL −1 in lake water and 0.2–2.8 pg mL −1 in milk, while recoveries were in the range of 72.3 %–124.7 % with RSD < 10.3 %. This approach provides an effective strategy for monitoring trace fluoroquinolones in environmental and food matrices, showcasing the potential of DBS-EB-COF-enhanced high-throughput SPME systems for applications in environmental and food safety surveillance.
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2017
·Vol. 409
·期 19
·4559-4569
·
IF 3.431
Xu Jia,Li Yunfang,Li Cao,Zhang Rui,Xiao Yuxiu
A hexafluoroisopropanol (HFIP)-mediated cloud point extraction (CPE) system was established. A small amount of HFIP (even 1%, v / v ) can dramatically reduce the cloud point of Triton X-100 (TX-100) aqueous solution (even to 1 °C) and make liquid-liquid two-phase separation (coacervate phase and aqueous phase) occur at room temperature over a wide range of TX-100 concentration (0.5∼10%, g/mL). HFIP-mediated coacervate phase has smaller volume (volume ratio is 1.8∼8.9% relative to the volume of the total solution with 1∼5% TX-100) and larger micelle aggregates (30∼80 nm in diameter) compared to temperature-induced coacervate phase (volume ratio at 2.8∼14.0%, the diameter of micelle aggregates at 5∼30 nm). HFIP-mediated CPE was coupled to high-performance liquid chromatography with ultraviolet detection (HPLC-UV) for the extraction and detection of organic pollutants in water, namely, polycyclic aromatic hydrocarbons (PAHs), fluoroquinolones (FQs), and sulfonamides (SAs) with different polarities, charges, and hydrogen-bonding properties. HFIP-mediated CPE provides much higher extraction rates (ERs) and enrichment factors (EFs) for FQs (91∼106%, 50∼59), PAHs (63∼90%, 33∼49), and SAs (26∼55%, 16∼34) compared with the temperature-induced one (ERs: 4∼8% for FQs, 25∼46% for PAHs, and 4∼37% for SAs; EFs: 1∼3 for FQs, 6∼12 for PAHs, and 8∼13 for SAs). The limit of detection ranges from 0.24 to 0.33 ng/mL for FQs, 0.04 to 0.38 ng/mL for PAHs, and 0.63 to 1.31 ng/mL for SAs. The proposed method was applied in the analysis of real water samples, and the recovery of 79.4∼110.8% and the relative standard deviation of 0.2∼16.3% were achieved for the three types of pollutants.
Schematic illustration of HFIP-mediated cloud point extraction
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高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Jing Xu,Si-Heng Luo,Chen-ru Xiong,Jia-hao Liu,Zeyu Zhao,Wei-qi Lin,Wei Zhang,Ping Guo,Cheng Pan,Quanlong Li,Zhong-Qun Tian,Bin Ren,Guo-Kun Liu
The overused quinolone antibiotics in animal husbandry and clinical medicine pose a growing threat for global health as they enter ecosystem via agricultural discharge and medical wastewater. Consequently, risk assessment for environmental and human exposure is highly demanded, which calls for an accurate, reliable and rapid qualitative and quantitative analysis of trace antibiotics in different matrices. Surface-enhanced Raman spectroscopy (SERS), providing fingerprint chemical information with near-single-molecule sensitivity, has received considerable attention. Herein, considering the common physicochemical properties of various quinolone antibiotics, we first developed sample pretreatment strategy consisting of solid-phase extraction (SPE), liquid–liquid extraction (LLE), salting-out assisted liquid–liquid extraction (SALLE), and NPs purification (NPs (p)). The modular SPE–LLE–SALLE–NPs (p) coupling with spectral separate network (SSNet) (a deep-learning based spectral unmixing algorithm) enabled a rapid (<8 min per sample), sensitive (μg L–1), on-site, and intelligent SERS analysis of 19 quinolone antibiotics in food, biological, and environmental matrices. Besides, the identification of each QN in a five-target mixture sample enabled SPE–LLE–SALLE–NPs (p)–SSNet–SERS to be a highly promising rapid detection strategy in blind sample screening qualitatively and quantitatively. The integrated SPE–LLE–SALLE–NPs (p)–SSNet–SERS offers a cross-media, full-process, and big data-enhanced rapid monitoring solution for antibiotic pollution within the “source-pathway-sink” framework.
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高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2025
·
IF 4.6
Xiang Li,Lei Jia,Yongxin Li,Bing Bi,Lina Zhang,Jun Xu,Tongqian Zhao
The residues of widely used fluoroquinolone antibiotics (FQs), including danofloxacin (DNX), enrofloxacin (ENR), norfloxacin (NFX), and pefloxacin (PEF), may accumulate in humans through contaminated water/food, posing health risks. Therefore, the rapid detection of FQs is of great research significance. In light of this, Tb/CdTe QD probes were developed in this study for the quantitative detection of multiple FQs, and a logic gate sensor array was constructed based on this probe to enable qualitative differentiation of these antibiotics. The probe possessed excellent anti-interference capability and remarkable stability. In real samples detection, such as lake water, tap water, milk, honey, and eggs, high recovery rates were successfully achieved. The detection limits for DNX, ENR, NFX, and PEF were 3.69 nM, 6.56 nM, 8.70 nM, and 4.72 nM, respectively. Furthermore, by integrating this ratio fluorescent probe with the color recognition software on a smartphone, visual semi-quantitative detection of FQs was successfully realized.
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高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2024
·
IF 4
Chang Liu,Yuan Chen,Hong Lin,Limin Cao,Kaiqiang Wang,Xiudan Wang,Jianxin Sui
ABSTRACT Low specificity and sensitivity often restrict the fluoroquinolones (FQs) detection when traditional IgG antibodies are used as recognition elements in immunoassay. Herein, shark-derived single-domain antibodies (ssdAbs) were used as new recognition elements to enhance the sensitivity and specificity for norfloxacin detection in immunoassay. Firstly, NOR-specific clones were isolated by phage display technology. Finally, five clones were successfully isolated from a NOR-immunized library. Secondly, a soluble expression system was constructed to produce ssdAbs. The 1N9 ssdAb exhibits the best affinity to the NOR complete antigen, with a limit of detection of 0.392 ng/mL and the cross-reaction rates of less than 8.240%, indicating its high sensitivity and specificity as a new recognition element in NOR detection. More importantly, the ELISA result proved that in NOR-negative fish matrix, no serious interference was observed, the 1N9 ssdAb could still recognize complete antigen NOR-OVA effectively. While in icELISA, 1N9 ssdAb could recognize NOR in different NOR-spiked concentrations between 10 to 1000 ng/mL, with the recovery rates between 93.66 - 116.49% with high reproducibility. Our study provides a new and effective recognition element with great potential for NOR detection.
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高影响因子
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