JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2025
·
IF 2.8
Xiao Ding,Shuo Wang,Wei Wang,Jingjing Sun,Chengcheng Jiang,Jianhua Hao
Recent studies on immunoaffinity chromatography (IAC) columns for aflatoxin B 1 (AFB 1 ) have not provided a comprehensive preparation procedure or explored their capacity, reusability, and stability. In this study, we outline a preparation process for an AFB 1 immune affinity column filler (AFB 1 -IAC) for extracting AFB 1 from food sources like rice, peanuts, and soybeans. Using commercially available IAC is often limited due to high costs, low capacity, slow flow rates, and poor reusability. To overcome these issues, we developed succinic anhydride (SA)-modified agarose pre-crosslinked microspheres (SA-A) activated with 1,1′‑carbonyldiimidazole (CDI). The activated microspheres were coupled with the anti-AFB 1 nanobody Nb02, creating an immunoaffinity packing called CDI-SA-A-Nb02 (D1-IAC). After verification, the results indicated that the column capacity of D1-IAC was about 4.67 times that of several commercial IAC brands. Additionally, the maximum flow rate at a pressure of 0.04 MPa was 25 % greater than specific commercial IAC, and the recovery rate remained above 80 % after being reused five times. This method is considered effective for extracting and detecting AFB 1 in food products. The preparation process for D1-IAC shows great consistency, and the column's performance is better than that of available immunoaffinity columns. This study explores using an IAC to extract and purify AFB 1 , which is then quantified using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).
研究领域:
食品安全
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生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2016
·
IF 2.687
Li Li,Li-Ru Xia,Yong-Fu Zhao,He-Ye Wang
In this paper, a novel highly sensitive chemiluminescence immune-affinity 96 spots monolith array was developed to detect deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and fumonisin B 1 (FB 1 ) in corn samples. Firstly, the monolith array was prepared through on suit UV-initiated copolymerization using polyethylene glycol diacrylate (PEGDA) as cross-linker, glycidyl methacrylate (GMA) as functional monomer and polyethylene glycol 200 (PEG 200) as the porogen. Subsequently, the four mycotoxins immune-affinity monolith array was prepared by immobilization of DON, ZEN, T-2, and FB 1 antibody. The mole ratio of PEGDA/GMA, UV exposure time, and the volume ratio of PEG 200/PEGDA were optimized to improve the performances of the immune-affinity monolith array. For the mycotoxins immune-affinity monolith array based on chemiluminescence detection, the limit of detection was 0.0036 ng/mL (DON), 0.0048 ng/mL (ZEN), 0.0039 ng/mL (T-2), and 0.0017 ng/mL (FB 1 ), respectively. The linear response in the range of 0.01–0.1 ng/mL (R 2 = 0.98). The results showed that the proposed four mycotoxins immune-affinity monolith array was a stable, accurate, and highly sensitive method to determine levels of DON, ZEN, T-2, and FB 1 in real samples.
研究领域:
食品科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2018
·Vol. 410
·期 8
·2253-2262
·
IF 3.307
Li Li,Chen Hongpu,Lv Xiaolan,Wang Min,Jiang Xizhi,Jiang Yifei,Wang Heye,Zhao Yongfu,Xia Liru
Mycotoxins produced by different species of fungi may coexist in cereals and feedstuffs, and could be highly toxic for humans and animals. For quantification of multiple mycotoxins in cereals, we developed a paper-based mycotoxin immune-affinity array. First, paper-based microzone arrays were fabricated by photolithography. Then, monoclonal mycotoxin antibodies were added in a copolymerization reaction with a cross-linker to form an immune-affinity monolith on the paper-based microzone array. With use of a competitive immune-response format, paper-based mycotoxin immune-affinity arrays were successfully applied to detect mycotoxins in samples. The detection limits for deoxynivalenol, zearalenone, T-2 toxin, and HT-2 toxin were 62.7, 10.8, 0.36, and 0.23 μg·kg -1 , respectively, which meet relevant requirements for these compounds in food. The recovery rates were 81–86% for deoxynivalenol, 89–117% for zearalenone, 79–86% for T-2 toxin, and 78–83% for HT-2 toxin, and showed the paper-based immune-affinity arrays had good reproducibility. In summary, the paper-based mycotoxin immune-affinity array provides a sensitive, rapid, accurate, stable, and convenient platform for detection of multiple mycotoxins in agro-foods.
Paper-based immune-affinity monolithic array. DON deoxynivalenol, HT-2 HT-2 toxin, T-2 T-2 toxin, PEGDA polyethylene glycol diacrylate, ZEN zearalenone
研究领域:
食品科学
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免疫学
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分析化学
✓ 已验证
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2022
·
IF 9.231
Peng Zhai,Chen Liu,Gang Feng,Yuanhao Cao,Lei Xiang,Keshi Zhou,Ping Guo,Jianqing Li,Wenxiao Jiang
Despite decades of efforts, we are faced with the daunting task of on-site ultratrace environmental toxins detecting, especially the microcystins caused by water bloom. In this work, a novel fluorescent microsphere-based flow-through immunoaffinity chromatographic assay has been designed for detecting ultratrace microcystin-LR in water and aquatic products. The aggregation-induced emission luminogens were encapsulated into fluorescent microspheres to ensure microcystin-LR quantitation with a whole analytical time of less than 30 min. Furthermore, the colorimetric images were captured and quantitatively analyzed, which offered a limit of detection at 0.217 pg/mL and a limit of quantitation at 0.362 pg/mL in water and aquatic muscle samples. The developed immunoassays provide average recovery ranging from 79.1 % to 95.7 %, with relative standard deviations less than 13.4 %. Thus, the validated flow-through immunoaffinity chromatographic assay is an easy-to-use alternative for on-site screening of microcystin-LR in water and aquatic samples at picogram levels.
研究领域:
环境科学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Food Measurement and Characterization
·
2024
·
IF 2.9
Tian Ye,Liu Zhaoying,Sun Mingna,Duan Jinsheng,Verheecke-Vaessen Carol,Du Daolin,Li Ming
To further promote the pretreatment technique, the immunoaffinity column (IAC), immunoaffinity magnetic (IAM) and multi-component immunoaffinity magnetic (multi-IAM) for mycotoxins were developed and evaluated systematically. Anti-mycotoxin-McAbs had immobilized on the CNBr-Sepharose 4B, carboxyl magnetic bead and protein G magnetic bead to prepare the IAC/IAM/multi-IAM, respectively. The capture efficiencies of mycotoxins (100 ng) reached 92.8% for IAC and exceeded 94.0% for IAM/multi-IAM. The multi-IAM showed more beneficial features in multiple targets, pretreatment time and recycling time than IAC/IAM, which could simultaneously pretreat AFB 1 /DON/ZEN in 15 min with three recycling times. The recoveries of IAC/IAM/multi-IAM coupled with ELISA ranged from 85.2 to 105.1%, with RSD between 4.0 and 13.2%. Moreover, the mycotoxin-positive authentic samples were detected from 1.1 to 749.6 ng/g by ELISA, and 1.2 to 762.7 ng/g by LC-MS/MS, with correlated R 2 exceeding 0.9955. The proposed IAC/IAM/multi-IAM owned the desired performance, such as specificity, accuracy, simpleness, efficiency, cost-effectiveness, simultaneity and environmentally friendly. This study can provide the theoretical basis for three immunoaffinity pretreatment techniques systematically, which may guide their application under different requirements and scenarios. Graphical abstract
研究领域:
食品科学
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分析化学
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2018
·
IF 2.441
Kai Yao,Kai Wen,Wenchong Shan,Sanlei Xie,Tao Peng,Jianyi Wang,Haiyang Jiang,Bing Shao
An immunoaffinity clean-up material based on a monoclonal antibody (mAb) has been prepared for concentrating and purifying bisphenol A (BPA) in 14 kinds of foodstuffs at trace level. Haptens and immunogen of bisphenol A have been synthesized and comprehensively characterized. An mAb towards BPA was prepared and cross-reactivities with 14 BPA analogues were below 5%. The prepared antibody was coupled to N -hydroxysuccinimide-activated Sepharose 4B to manufacture an immunoaffinity column (IAC), which was applied to purify BPA in 14 kinds of foodstuffs. The analyte was then detected by means of ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Under the optimized conditions, compared with two traditional SPE clean-up methods, the IAC showed better selectivity (matrix effect <16.8%) and higher sensitivity. The limits of detection for BPA in 14 kinds of foodstuffs ranged from 0.001 μg L −1 to 0.01 μg kg −1 , and the limits of quantification were in the range from 0.003 μg L −1 to 0.04 μg kg −1 . The recoveries of BPA from spiked samples ranged from 82.0% to 104.9%, with RSDs below 13.8%. Besides, the IAC exhibited good reusability, with 40% column capacity remaining and no significant loss of recovery after 25 application cycles in real sample detection. These results demonstrated that the developed IAC-UPLC-MS/MS approach has wide applicability for purifying and detecting BPA in various foodstuffs.
研究领域:
分析化学
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食品安全
·
环境科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2013
·
IF 2.582
Qin Zhang,Xiuwen Tang,Fenghua Hou,Jianping Yang,Zhiyong Xie,Zhiyi Cheng
We fabricated a three-layer polydimethylsiloxane (PDMS)-based microfluidic chip for realizing urease inhibition assay with sensitive fluorescence detection. Procedures such as sample prehandling, enzyme reaction , reagent mixing, fluorescence derivatization , and detection can be readily carried out. Urease reactors were prepared by adsorption of rabbit immunoglobulin G (IgG) and immunoreaction with urease-conjugated goat anti-rabbit IgG. Acetohydroxamic acid (AHA) as a competitive inhibitor of urease was tested on the chip. Microfluidically generated gradient concentrations of AHA with substrate (urea) were loaded into urease reactors. After incubation, the produced ammonia was transported out of reactors and then reacted with o -phthalaldehyde (OPA) to generate fluorescent products. Urease inhibition was indicated by a decrease in fluorescence signal detected by microplate reader. The IC 50 value of AHA was determined and showed good agreement with that obtained in microplate. The presented device combines several steps of the analytical process with advantages of low reagent consumption, reduced analysis time, and ease of manipulation. This microfluidic approach can be extended to the screening of inhibitory compounds in drug discovery.
研究领域:
生物化学
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药物开发
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分析化学
✓ 已验证
DOI
PubMed
高影响因子
Analytical Methods
·
2019
·Vol. 11
·期 48
·6084-6091
·
IF 2.378
He-Ye Wang,Ya-Li Yang,Yue Wang,Shuai-Feng Zhang,Li Li
Organic polymer monoliths have been used extensively for separation and purification of molecules because of their excellent mass transfer properties and versatile surface modification. However, organic polymer monolithic materials have significant UV absorption, which blocked UV-visible absorbance detection. Thus, we developed novel disk-shaped mycotoxin immune-affinity monolithic microplates for quantitative detection of mycotoxins. The concentric disk-shaped format allows the obscure organic monoliths to capture mycotoxins and unobstructed UV-visible absorbance measurement was performed in the same microplates. The limits of detection of 96 disk-shaped mycotoxin immune-affinity monolithic microplates were 161.73, 22.52, 0.14, and 0.21 μg kg−1 for Deoxynivalenol (DON), Zearalenone (ZEN), Aflatoxin B1 (AFB1), and Ochratoxin A (OTA), respectively, which meet well the requirements for detection of foodstuffs. The recovery rate ranges of the four mycotoxins were 87–92%, 82–89%, 78–116%, and 83–87% for DON, ZEN, AFB1, and OTA disk-shaped microplates, respectively, which proved that the disk-shaped monolithic microplates have good reproducibility for mycotoxin detection. In summary, 96 disk-shaped mycotoxin immune-affinity monolithic microplates are a sensitive, accurate, stable, and convenient platform for detection of mycotoxins in agro-food.
研究领域:
分析化学
·
食品安全
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2023
·
IF 4.8
Fatemeh Kardani,Roya Mirzajani,Yousef Tamsilian,Alireza Kiasat,Farnoosh Bakhshandeh Farajpour
In the present study, for the first time, immunoaffinity columns under the term MIL (Al) −53 deep eutectic solvents @MIPs (MIL(Al)-53-DES@MIPs) were synthesized based on two technologies, metal–organic framework (MOF) and molecular imprinted polymers (MIPs). The MOFs and MIPs with DES used in synthesizing immunoaffinity columns, was subsequently used to extract and preconcentrate aflatoxins AFB1, AFB2, AFG1, and AFG2. One factor at a time (OFAT) method was used to optimize the parameters affecting on extraction efficiency of the target compounds. Under optimized conditions, the limits of detection of the four aflatoxin ranged from 0.023 to 0.033 µg kg −1 (S/N = 5), and the calibration graphs were linear in the concentration range from 0.1 to 400 μg kg −1 , the inter-day and intraday relative standard deviations (RSD) for various aflatoxins at three different concentration level (n = 5) using a single column were 1.3–4.4 %, and the column to column RSD % (n = 5) was 4.3–6.7 % at 5 μg L -1 of each analyte. The method was successfully applied to quantify aflatoxin in cereals samples of Khuzestan province of Iran, with the recoveries between 95.3 and 98.5 %.
研究领域:
食品安全
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2021
·
IF 3.205
Dan-Tong Zhao,Yi-Jun Gao,Wen-Jin Zhang,Tian-Chen Bi,Xiao Wang,Chun-Xia Ma,Rong Rong
Mycotoxins are poisonous secondary fungal toxic metabolites and harmful to human health. Traditional Chinese medicinal materials (TCMs), including more than two hundred functional foods, are vulnerably bred fungi, causing spoilage and multi-mycotoxins contamination. This study established a simultaneous analytical method by using multi-mycotoxins immunoaffinity column (multi-IAC) and HPLC-MS/MS to evaluate mycotoxins' contamination levels and natural incidence in TCMs. Aflatoxins (AFs, including AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), fumonisins (FB1 and FB2), zearalenone (ZEN), deoxynivalenol (DON) and T-2 toxins in three TCMs or functional foods of Polygalae Radix (PR), Coicis Semen (CS) and Eupolyphaga Steleophaga (ES) were detected. The systematically investigated results of 30 batch AFB1 positive samples revealed co-occurrence and correlation of multi-mycotoxins are significant differences in various matrices. All the samples in this study contain more than 5 mycotoxins. AFB1-AFs, AFB1-FBs, AFB1-DON, and AFB1-T-2 are the most observed co-occurrence, AFB1-OTA is also of concern due to its synergistic toxicity. This study's results can be used to establish guidelines for screening mycotoxin contaminants and limitations on acceptable levels in TCMs. Simultaneously, mycotoxin's correlation results in different matrices can also provide a reference for the standardization of TCM production and processing.
研究领域:
食品科学
·
分析化学
·
毒理学
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DOI
PubMed
高影响因子
Journal of Physical Chemistry C
·
2022
·
IF 4.177
Yun Ling,Ya-ru Chu,Feng Gao,Qing-yun Feng,Hui-qi Xie,Yong Shao,Qing-xiang Wang
The unique chemical and physical properties of metal–organic frameworks (MOFs) endow them with promising applications in the electrochemical sensing field. Herein, a novel label-free electrochemical sensing strategy for cardiac troponin I (cTnI) has been constructed on the basis of immunoaffinity-induced depression of the electrochemical signal of Cu-MOF-74. The electrochemical assay shows that the immunoaffinity reaction between cTnI and anti-cTnI on the electrode surface can effectively inhibit the electrochemical signal of electrode-confined Cu-MOF-74, suggesting that the Cu-MOF-74-based interface may find application in the label-free analysis of cTnI. Thus, Cu-MOF-74 acts as a bifunctional material in this strategy, namely, the immobilization matrix of probe molecules and the signal source of the sensing analysis. In addition, the reason for the electrochemical signal depression of Cu-MOF-74 induced by immunoaffinity reaction is attributed to the increase in the insulation layer and the blocking of the electrolyte diffusion from the bulk solution to the electrode surface. In this strategy, the analytical range of cTnI is from 0.01 pg mL–1 to 1.0 ng mL–1 and the detection limit is 4.5 fg mL–1. The Cu-MOF-74-based sensing interface is also reliable for cTnI analysis in real serum samples, showing that the biosensor has promising application in point-of-care testing (POCT) of myocardial injury-related diseases.
研究领域:
生物传感器
·
医学诊断
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
Foods
·
2022
·Vol. 11
·期 3
·335
·
IF 5.561
Disha Lu,Xu Wang,Ruijue Su,Yongjian Cheng,Hong Wang,Lin Luo,Zhili Xiao
A novel and efficient immunoaffinity column (IAC) based on bispecific monoclonal antibody (BsMAb) recognizing aflatoxin B1(AFB1) and ochratoxin A (OTA) was prepared and applied in simultaneous extraction of AFB1and OTA from food samples and detection of AFB1/OTA combined with ic-ELISA (indirect competitive ELISA). Two deficient cell lines, hypoxanthine guanine phosphoribosyl-transferase (HGPRT) deficient anti-AFB1hybridoma cell line and thymidine kinase (TK) deficient anti-OTA hybridoma cell line, were fused to generate a hybrid-hybridoma producing BsMAb against AFB1and OTA. The subtype of the BsMAb was IgG1via mouse antibody isotyping kit test. The purity and molecular weight of BsMAb were confirmed by SDS-PAGE method. The cross-reaction rate with AFB2was 37%, with AFG115%, with AFM148%, with AFM210%, and with OTB 36%. Negligible cross-reaction was observed with other tested compounds. The affinity constant (Ka) was determined by ELISA. The Ka (AFB1) and Ka (OTA) was 2.43 × 108L/mol and 1.57 × 108L/mol, respectively. Then the anti-AFB1/OTA BsMAb was coupled with CNBr-Sepharose, and an AFB1/OTA IAC was prepared. The coupling time and elution conditions of IAC were optimized. The coupling time was 1 h with 90% coupling rate, the eluent was methanol–water (60:40,v:v, pH 2.3) containing 1 mol/L NaCl, and the eluent volume was 4 mL. The column capacities of AFB1and OTA were 165.0 ng and 171.3 ng, respectively. After seven times of repeated use, the preservation rates of column capacity for AFB1and OTA were 69.3% and 68.0%, respectively. The ic-ELISA for AFB1and OTA were applied combined with IAC. The IC50(50% inhibiting concentration) of AFB1was 0.027 ng/mL, the limit of detection (LOD) was 0.004 ng/mL (0.032 µg/kg), and the linear range was 0.006 ng/mL~0.119 ng/mL. The IC50of OTA was 0.878 ng/mL, the LOD was 0.126 ng/mL (1.008 µg/kg), and the linear range was 0.259 ng/mL~6.178 ng/mL. Under optimum conditions, corn and wheat samples were pretreated with AFB1-OTA IAC. The recovery rates of AFB1and OTA were 95.4%~105.0% with ic-ELISA, and the correlations between the detection results and LC-MS were above 0.9. The developed IAC combined with ic-ELISA is reliable and could be applied to the detection of AFB1and OTA in grains.Keywords:aflatoxin B1;ochratoxin A;bispecific monoclonal antibody;immunoaffinity column;ic-ELISA
研究领域:
食品科学
·
分析化学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）