Europium Fluorescent Nanoparticles-Based Multiplex Lateral Flow Immunoassay for Simultaneous Detection of Three Antibiotic Families Residue

A fluorescent immunoassay based on europium nanoparticles (EuNPs-FIA) was developed for the simultaneous detection of antibiotic residues, solving the problems of single target detection and low sensitivity of traditional immunoassay methods. In the EuNPs-FIA, EuNPs were used as indictive probes by binding to anti-tetracyclines monoclonal antibodies (anti-TCs mAb), anti-sulphonamides monoclonal antibodies (anti-SAs mAb) and anti-fluoroquinolones monoclonal antibodies (anti-FQs mAb), respectively. Different artificial antigens were assigned to different regions of the nitrocellulose membrane as capture reagents. The EuNPs-FIA allowed for the simultaneous detection of three classes of antibiotics (tetracyclines, fluoroquinolones and sulphonamides) within 15 min. It enabled both the qualitative determination with the naked eye under UV light and the quantitative detection of target antibiotics by scanning the fluorescence intensity of the detection probes on the corresponding detection lines. For qualitative analysis, the cut-off values for tetracyclines (TCs), fluoroquinolones (FQs) and sulphonamides (SAs) were 3.2 ng/ml, 2.4 ng/ml and 4.0 ng/ml, respectively, which were much lower than the maximum residue limit in food. For quantitative analysis, these ranged from 0.06 to 6.85 ng/ml for TCs, 0.03–5.14 ng/ml for FQs, and 0.04–4.40 ng/ml for SAs. The linear correlation coefficients were higher than 0.97. The mean spiked recoveries ranged from 92.1 to 106.2% with relative standard deviations less than 8.75%. Among them, the three monoclonal antibodies could recognize four types of TCs, seven types of FQs and 13 types of SAs, respectively, and the detection range could cover 24 antibiotic residues with different structural formulations. The results of the detection of antibiotic residues in real samples using this method were highly correlated with those of high performance liquid chromatography (R2 > 0.98). The accuracy and precision of the EuNPs-FIA also met the requirements for quantitative analysis. These results suggested that this multiplex immunoassay method was a promising method for rapid screening of three families of antibiotic residues.

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Duplex Surface Enhanced Raman Scattering-Based Lateral Flow Immunosensor for the Low-Level Detection of Antibiotic Residues in Milk

Molecules ◽

10.3390/molecules25225249 ◽

2020 ◽

Vol 25 (22) ◽

pp. 5249

Author(s):

Ruiqi Fan ◽

Shusheng Tang ◽

Sunlin Luo ◽

Hu Liu ◽

Wanjun Zhang ◽

...

Keyword(s):

Raman Scattering ◽

Simultaneous Detection ◽

Surface Enhanced Raman Scattering ◽

Lateral Flow ◽

Antibiotic Residues ◽

Relative Standard ◽

Surface Enhanced ◽

Surface Enhanced Raman ◽

Enhanced Raman Scattering ◽

Analytical Approaches

A duplex surface enhanced Raman scattering (SERS)-based lateral flow immunosensor was established for the simultaneous detection of two common antibiotic residues including tetracycline and penicillin in milk. The newly synthesized Au@Ag nanoparticles were labeled with different Raman molecules including 5,5-dithiobis-2-nitrobenzoic acid (DTNB) or 4-mercaptobenzoic acid (MBA), followed by the conjugation of anti-tetracycline monoclonal antibody or anti-penicillin receptor, forming two kinds of SERS nanoprobes. The two nanoprobes can recognize tetracycline-BSA and ampicillin-BSA, respectively, which facilitates the simultaneous detection of the two types of antibiotics on a single test line. After optimization, detection limits of tetracycline and penicillin as low as 0.015 ng/mL and 0.010 ng/mL, respectively, were achieved. These values were far below those of most of other documented bio-analytical approaches. Moreover, the spiking test demonstrates an excellent assay accuracy with recoveries of 88.8% to 111.3%, and satisfactory assay precision with relative standard deviation below 16%. Consequently, the results demonstrate that the SERS-based lateral flow immunosensor developed in this study has the advantages of excellent assay sensitivity and remarkable multiplexing capability, thus it will have great application potential in food safety monitoring.

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Development of a Direct Competitive ELISA Kit for Detecting Deoxynivalenol Contamination in Wheat

Molecules ◽

10.3390/molecules25010050 ◽

2019 ◽

Vol 25 (1) ◽

pp. 50

Author(s):

Li Han ◽

Yue-Tao Li ◽

Jin-Qing Jiang ◽

Ren-Feng Li ◽

Guo-Ying Fan ◽

...

Keyword(s):

High Throughput Screening ◽

High Performance ◽

Limit Of Detection ◽

High Specificity ◽

Hplc Method ◽

Rapid Screening ◽

Enzyme Linked Immunosorbent Assay ◽

Detection Range ◽

Technical Requirements ◽

Relative Standard

This study was conducted to develop a self-assembled direct competitive enzyme-linked immunosorbent assay (dcELISA) kit for the detection of deoxynivalenol (DON) in food and feed grains. Based on the preparation of anti-DON monoclonal antibodies, we established a standard curve with dcELISA and optimized the detection conditions. The performance of the kit was evaluated by comparison with high-performance liquid chromatography (HPLC). The minimum detection limit of DON with the kit was 0.62 ng/mL, the linear range was from 1.0 to 113.24 ng/mL and the half-maximal inhibition concentration (IC50) was 6.61 ng/mL in the working buffer; there was a limit of detection (LOD) of 62 ng/g, and the detection range was from 100 to 11324 ng/g in authentic agricultural samples. We examined four samples of wheat bran, wheat flour, corn flour and corn for DON recovery. The average recovery was in the range of 77.1% to 107.0%, and the relative standard deviation (RSD) ranged from 4.2% to 11.9%. In addition, the kit has the advantages of high specificity, good stability, a long effective life and negligible sample matrix interference. Finally, wheat samples from farms in the six provinces of Henan, Anhui, Hebei, Shandong, Jiangsu and Gansu in China were analyzed by the kit. A total of 30 samples were randomly checked (five samples in each province), and the results were in good agreement with the standardized HPLC method. These tests showed that the dcELISA kit had good performance and met relevant technical requirements, and it had the characteristics of accuracy, reliability, convenience and high-throughput screening for DON detection. Therefore, the developed kit is suitable for rapid screening of DON in marketed products.

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Sample Preincubation Strategy for Sensitive and Quantitative Detection of Clenbuterol in Swine Urine Using a Fluorescent Microsphere–Based Immunochromatographic Assay

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-086 ◽

2014 ◽

Vol 77 (11) ◽

pp. 1998-2003 ◽

Cited By ~ 18

Author(s):

SHENG L. DENG ◽

SHAN SHAN ◽

CHAO L. XU ◽

DAO F. LIU ◽

YONG H. XIONG ◽

...

Keyword(s):

Limit Of Detection ◽

Lateral Flow ◽

Lateral Flow Assay ◽

Rapid Screening ◽

Immunochromatographic Assay ◽

Quantitative Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescent Microsphere ◽

Swine Urine ◽

Relative Standard

We describe an ultrasensitive and quantitative immunochromatographic assay to determine the amount of clenbuterol (CLB) in swine urine. In this study, fluorescein isothiocyanate polystyrene fluorescent microspheres were used as probes. A sample preincubation strategy was introduced to this immunochromatographic assay. Results showed that the strategy evidently improved the sensitivity and accuracy of lateral flow assay. The method was completed in 20 min, and a half-maximal inhibitory concentration of 0.13 μg liter−1 was obtained. The limit of detection of the proposed method to determine CLB in swine urine was 0.01 μg liter−1, which was lower than the limit of detection of immunochromatographic assays without preincubation. Intra-and interday recoveries of spiked swine urine ranged from 85.0 to 107.5%. The relative standard deviation values of the preincubated test strip ranged from 2.7 to 12.5%. Analysis of the CLB in swine urine samples showed that the result obtained from the lateral flow assay is consistent with that obtained from a commercial enzyme-linked immunosorbent assay kit. Our results suggest that the developed fluorescent microsphere–based immunochromatographic assay may be useful as a rapid screening method to detect CLB quantitatively.

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Screening of 49 antibiotic residues in aquatic products using modified QuEChERS sample preparation and UPLC-QToFMS analysis

10.7717/peerj-achem.8 ◽

2021 ◽

Vol 3 ◽

pp. e8

Author(s):

Yao Gao ◽

Tianwen Zhang ◽

Shirong Huang ◽

Xinxin Lin ◽

Sisi Gong ◽

...

Keyword(s):

Sample Preparation ◽

Grass Carp ◽

High Performance ◽

Gradient Elution ◽

Rapid Screening ◽

Scylla Serrata ◽

Penaeus Vannamei ◽

Antibiotic Residues ◽

Aquatic Products ◽

Relative Standard

A precise analytical method was established for rapid screening of 49 antibiotic residues in aquatic products by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QToFMS). The quick, easy, cheap, effective, rugged and safe (QuEChERS) process was refined for effective sample preparation. The homogenized samples of aquatic products were extracted with 3% acetic acid in acetonitrile, salted out with anhydrous magnesium sulfate and sodium chloride, and cleaned up by octadecylsilane (C18) and primary-secondary amine (PSA) powder. Then, the purified samples were separated on a BEH C18 column using 0.1% formic acid and methanol as mobile phases by gradient elution, detected by MS under positive Electron Spray Ionization (ESI+) mode. The linear range of matrix-matched calibration curve was 1–100 μg/L for each compound with the correlation coefficients in the range of 0.9851–0.9999. The recoveries of target antibiotics at the different spiked levels ranged from 60.2% to 117.9% except for lincomycin hydrochloride, whereas relative standard deviations (RSDs) were between 1.6% and 14.0% except for sulfaguanidine in grass Carp, Penaeus vannamei and Scylla serrata matrices. The limits of detection (LODs) (S/N = 3) for the analytes were 0.05–2.40 μg/kg, 0.08–2.00 μg/kg and 0.10–2.27 μg/kg and the limits of quantification (LOQs) (S/N = 10) were 0.16–8.00 μg/kg, 0.25–6.66 μg/kg and 0.32–7.56 μg/kg in grass Carp, Penaeus vannamei and Scylla serrata, respectively. The method was successfully applied to grass Carp, Penaeus vannamei and Scylla serrata, demonstrating its ability for the determination of multi-categories antibiotic residues in aquatic products.

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Detection of Brevetoxin in Human Plasma by ELISA

Journal of Analytical Toxicology ◽

10.1093/jat/bkab010 ◽

2021 ◽

Author(s):

Brady R Cunningham ◽

Rebecca M Coleman ◽

Adam M Schaefer ◽

Elizabeth I Hamelin ◽

Rudolph C Johnson

Keyword(s):

Human Plasma ◽

Limit Of Detection ◽

Cross Reactivity ◽

Quantitative Detection ◽

Red Tides ◽

Plasma Samples ◽

Detection Range ◽

Paralytic Shellfish ◽

Test Kit ◽

Relative Standard

Abstract Florida red tides have become more common and persistent in and around the Gulf of Mexico. When in bloom, red tides can produce brevetoxins in high concentrations, leading to human exposures primarily through contaminated food and ocean spray. The research described here includes adapting and validating a commercial brevetoxin water test kit for human plasma testing. Pooled plasma was fortified with a model brevetoxin, brevetoxin 3, at concentrations from 0.00500 to 3.00 ng/mL to generate calibration curves and quality control samples. The quantitative detection range was determined to be 0.0400–2.00 ng/mL brevetoxin 3 equivalents with inter- and intraday accuracies ranging from 94.0% to 109% and relative standard deviations <20%, which is within the US Food and Drug Administration guidelines for receptor-binding assays. Additionally, cross-reactivity was tested using 4 of the 10 known brevetoxins and 12 paralytic shellfish toxins. The cross-reactivity varied from 0.173% to 144% for the commercially available brevetoxin standards and 0% for the commercially available paralytic shellfish toxin standards. Fifty individual unexposed human plasma samples were measured to determine the limit of detection and endogenous interferences to the test. The validated method was used to test 31 plasma samples collected from humans potentially exposed to brevetoxins, detecting 11 positives. This method has been proven useful to measure human exposure to brevetoxins and can be applied to future exposure events.

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Quantitative Detection of Chromium Pollution in Biochar Based on Matrix Effect Classification Regression Model

Molecules ◽

10.3390/molecules26072069 ◽

2021 ◽

Vol 26 (7) ◽

pp. 2069

Author(s):

Mei Guo ◽

Rongguang Zhu ◽

Lixin Zhang ◽

Ruoyu Zhang ◽

Guangqun Huang ◽

...

Keyword(s):

Regression Model ◽

Matrix Effect ◽

Nearest Neighbor ◽

Quantitative Detection ◽

Least Squares Regression ◽

K Nearest Neighbor ◽

Breakdown Spectroscopy ◽

Relative Standard ◽

Calibration Samples ◽

Standard Deviations

Returning biochar to farmland has become one of the nationally promoted technologies for soil remediation and improvement in China. Rapid detection of heavy metals in biochar derived from varied materials can provide a guarantee for contaminated soil, avoiding secondary pollution. This work aims first to apply laser-induced breakdown spectroscopy (LIBS) for the quantitative detection of Cr in biochar. Learning from the principles of traditional matrix effect correction methods, calibration samples were divided into 1–3 classifications by an unsupervised hierarchical clustering method based on the main elemental LIBS data in biochar. The prediction samples were then divided into diverse classifications of calibration samples by a supervised K-nearest neighbor (KNN) algorithm. By comparing the effects of multiple partial least squares regression (PLSR) models, the results show that larger numbered classifications have a lower averaged relative standard deviations of cross-validation (ARSDCV) value, signifying a better calibration performance. Therefore, the 3 classification regression model was employed in this study, which had a better prediction performance with a lower averaged relative standard deviations of prediction (ARSDP) value of 8.13%, in comparison with our previous research and related literature results. The LIBS technology combined with matrix effect classification regression model can weaken the influence of the complex matrix effect of biochar and achieve accurate quantification of contaminated metal Cr in biochar.

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Quantitative analysis of doxorubicin hydrochloride and arterolane maleate by mid IR spectroscopy using transmission and reflectance modes

BMC Chemistry ◽

10.1186/s13065-021-00752-3 ◽

2021 ◽

Vol 15 (1) ◽

Author(s):

Ranju Bansal ◽

Ranjit Singh ◽

Khushpal Kaur

Keyword(s):

Quantitative Analysis ◽

Spectroscopic Methods ◽

Doxorubicin Hydrochloride ◽

Intermediate Precision ◽

Solid Dosage ◽

Ich Guidelines ◽

Relative Standard ◽

Ft Ir ◽

Solid Bulk ◽

Ir Spectroscopic

Abstract Background Environment-friendly fast and accurate mid-infrared spectroscopic methods have been developed for the quantitative analysis of doxorubicin hydrochloride (DOX) and arterolane maleate (ALM) in bulk and marketed formulations. Both transmittance and reflectance modes have been used for the analysis and a comparison has been drawn for better accuracy. The analytical methods were validated in accordance with International Council for Harmonisation (ICH) guidelines Results The proposed methods have been successfully developed and validated for the quantification of doxorubicin and arterolane maleate in solid bulk and dosage form. High recovery values in both the modes, while analysing DOX and ALM, indicated good accuracy of the methods. The methods showed excellent repeatability and intermediate precision [% RSD (Relative Standard Deviation < 2.0%]. The assay values of the drugs in solid dosage forms were also found close to the labelled claim. Conclusion The proposed Fourier transform infrared (FT-IR) spectroscopic methods were found to be specific, reproducible, valid and could be used as general methods for the quantification of most of the solid drug preparations such as tablets, capsules and powders.

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Quantitative analysis of direct oral anticoagulant rivaroxaban by terahertz spectroscopy

The Analyst ◽

10.1039/d0an00268b ◽

2020 ◽

Vol 145 (11) ◽

pp. 3909-3915 ◽

Cited By ~ 2

Author(s):

Xu Wu ◽

Liping Wang ◽

Yan Peng ◽

Fang Wu ◽

Jiumei Cao ◽

...

Keyword(s):

Quantitative Analysis ◽

Oral Anticoagulant ◽

Terahertz Spectroscopy ◽

New Method ◽

Quantitative Detection ◽

Direct Oral Anticoagulant ◽

Qualitative And Quantitative

A new method for the qualitative and quantitative detection of direct oral anticoagulant rivaroxaban by terahertz spectroscopy.

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A highly sensitive octopus-like azobenzene fluorescent probe for determination of abamectin B1 in apples

Scientific Reports ◽

10.1038/s41598-021-84221-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Zhenlong Guo ◽

YiFei Su ◽

Kexin Li ◽

MengYi Tang ◽

Qiang Li ◽

...

Keyword(s):

Fluorescent Probe ◽

Limit Of Detection ◽

Quantitative Detection ◽

Fluorescence Signal ◽

Ionization Mass ◽

Limit Of Quantification ◽

Visible Absorption ◽

Esi Ms ◽

Relative Standard ◽

Ft Ir

AbstractThe development of detecting residual level of abamectin B1 in apples is of great importance to public health. Herein, we synthesized a octopus-like azobenzene fluorescent probe 1,3,5-tris (5′-[(E)-(p-phenoxyazo) diazenyl)] benzene-1,3-dicarboxylic acid) benzene (TPB) for preliminary detection of abamectin B1 in apples. The TPB molecule has been characterized by ultraviolet–visible absorption spectrometry, 1H-nuclear magnetic resonance, fourier-transform infrared (FT-IR), electrospray ionization mass spectroscopy (ESI-MS) and fluorescent spectra. A proper determination condition was optimized, with limit of detection and limit of quantification of 1.3 µg L−1 and 4.4 μg L−1, respectively. The mechanism of this probe to identify abamectin B1 was illustrated in terms of undergoing aromatic nucleophilic substitution, by comparing fluorescence changes, FT-IR and ESI-MS. Furthermore, a facile quantitative detection of the residual abamectin B1 in apples was achieved. Good reproducibility was present based on relative standard deviation of 2.2%. Six carboxyl recognition sites, three azo groups and unique fluorescence signal towards abamectin B1 of this fluorescent probe demonstrated reasonable sensitivity, specificity and selectivity. The results indicate that the octopus-like azobenzene fluorescent probe can be expected to be reliable for evaluating abamectin B1 in agricultural foods.

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Development of a multicolor upconversion fluorescence immunoassay for the simultaneous detection of thiamethoxam and dextran by magnetic separation

RSC Advances ◽

10.1039/d0ra07954e ◽

2021 ◽

Vol 11 (1) ◽

pp. 517-524

Author(s):

Chuanyong Li ◽

Wanlin Sun ◽

Lianrun Huang ◽

Nana Sun ◽

Xiude Hua ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Simultaneous Determination ◽

Magnetic Separation ◽

Simultaneous Detection ◽

Fluorescence Immunoassay ◽

Upconversion Fluorescence

The anti-thiamethoxam and anti-dextran monoclonal antibodies were prepared to develop a multicolor upconversion fluorescence immunoassay for the simultaneous determination of thiamethoxam (544 nm) and dextran (477 nm).

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