Co-reporter:Fei Xu, Wenxiao Jiang, Jie Zhou, Kai Wen, Zhanhui Wang, Haiyang Jiang, and Shuangyang Ding
Journal of Agricultural and Food Chemistry 2014 Volume 62(Issue 14) pp:3108-3113
Publication Date(Web):March 24, 2014
DOI:10.1021/jf405379r
Apramycin (APR) residue in food of animal origin can cause harmful effects on human health. In this study, a monoclonal antibody (mAb) was successfully produced using APR–BSA as immunogen, which was prepared by using the glutaraldehyde method. mAb 2A2 showed low cross-reactivity (<0.1%) with other aminoglycoside antibiotics, and its IC50 value was 0.35 ng/mL. On the basis of this mAb, a novel immunoassay in the format of an immunoaffinity test column (IATC) was developed. An immunoaffinity column filled with anti-APR antibody–Sepharose 4B gel was used as solid phase. APR in sample and HRP–APR conjugate compete with each other for the limited antibody on the column. The approach was able to give a naked-eye color signal for the detection of analyte. A blue color appears for negative results and no color for positive. The method was then successfully applied to the detection of APR in animal-origin food. To further evaluate the assay, direct competitive ELISA (dcELISA) based on the same antibody was developed for comparison in different aspects. Compared to the dcELISA, the detection time of IATC is shortened to 20 min, whereas a similar sensitivity for various samples was observed. The limits of detection (LOD) for raw milk, muscles, and livers are 3 ng/mL, 3 μg/kg, and 10 μg/kg, respectively.
Co-reporter:Jie Zhou, Kui Zhu, Fei Xu, Wenjun Wang, Haiyang Jiang, Zhanhui Wang, and Shuangyang Ding
Journal of Agricultural and Food Chemistry 2014 Volume 62(Issue 49) pp:12061-12066
Publication Date(Web):October 7, 2014
DOI:10.1021/jf5029416
The residue of lincomycin (LIN) in edible animal foodstuffs caused by the widespread use of veterinary drugs is in need of rapid, simple, and sensitive detection methods. The present work introduces a fluorescent microsphere immunoassay (FMIA) for detecting LIN in different samples based on the competitive immunoreaction on the chromatography test strip. The residues of LIN in different samples compete with bovine serum albumin (BSA) labeled LIN conjugates on the T-line to bind to the anti-LIN monoclonal antibody labeled fluorescent microspheres (FM–mAbs). Captured FM–mAbs on the T-line represent the fluorescent intensity, which is detected under UV light and quantified by a fluorescent reader. Under optimized conditions, the dynamic range is from 1.35 to 3.57 ng/mL, and the 50% inhibition concentration (IC50) is 2.20 ng/mL. This method has 4.4% cross-reactivity with clindamycin and negligible cross-reactivity (<0.1%) with other analogues. To reduce the matrix effects, a dilution method is used to pretreat the samples, and the recoveries range from 73.92 to 120.50% with coefficient of variations <21.76%. In comparison with the results of ELISA and colloidal gold immunoassay, FMIA has obvious advantages such as easy operation, time savings, high sensitivity and specificity, and broader prospect.
Co-reporter:Fei Xu;Wenjun Wang;Haiyang Jiang;Zhaopeng Wang;Zhanhui Wang
Food Analytical Methods 2014 Volume 7( Issue 8) pp:1619-1626
Publication Date(Web):2014 September
DOI:10.1007/s12161-014-9797-7
Plasticizer has attracted more and more attention in China for the past 3 years, especially in Taiwan district. In this study, an indirect competitive enzyme-linked immunosorbent assay (icELISA) has been developed for the determination of a plasticizer dibutyl phthalate (DBP) in white wine. Dibutyl 4-aminophthalate coupled with OVA was synthesized as an immunogen to produce polyclonal antibodies against DBP. The antibody exhibited negligible cross-reactivity with other related compounds. The influence of several physicochemical parameters, such as coating procedure, organic solvent, competitive reaction time, and pH was investigated. The limit of detection was 64.5 ng/mL, which was sensitive enough for a screening assay. The linear range was 64.5–1,606.2 ng/mL with a correlation coefficient (R2) of 0.996. The method was successfully applied to the determination of DBP in white wine. Recoveries were between 83.1 and 101.7 %. This immunoassay was highly specific, sensitive, rapid, simple, and suitable for DBP monitoring. The results obtained were compared with those obtained using gas chromatography–mass spectrometry (GC-MS), and a satisfied correlation coefficient of 0.928 was obtained by real sample detection.
Co-reporter:Jiang Haiyang;Wang Wenjun;Zhu Jinghui;Tao Xiaoqi;Li Jiancheng;Xia Xi;Wen Kai;Xu Fei;Wang Zhaopeng;Chen Min;Li Xiangmei;Wu Xiaoping;Wang Shien;Ding Shuangyang
Luminescence 2014 Volume 29( Issue 4) pp:393-400
Publication Date(Web):
DOI:10.1002/bio.2559
ABSTRACT
A chemiluminescent enzyme immunoassay (CLEIA) was compared to an ultraperformance liquid chromatography tandem mass spectroscopy (UPLC-MS/MS) procedure for the analysis of zeranol and its metabolites in bovine tissue samples. Apparent recoveries from fortified samples by both methods were comparable at 0.5–4.0 µg/kg and a significant correlation was obtained. For CLEIA analysis, hapten mimicking the analyte was first synthesized and conjugated with the carrier protein bovine serum albumin as the immunogen to produce monoclonal antibody. The obtained antibody showed extensive cross-reactivity toward zeranol metabolites (zearalanone). The limit of detection of CLEIA and UPLC-MS/MS was 0.05 µg/kg and 0.5 µg/kg, respectively. Recoveries of both methods for fortified samples were higher than 75.0% with the coefficient of variation less than 15%. These results indicated that the combination of screening with CLEIA and confirmation with UPLC-MS/MS for zeranol and its metabolites would be a reliable method for a large number of bovine samples. Copyright © 2013 John Wiley & Sons, Ltd.
Co-reporter:Zhiming Xiao, Yunxia Yang, Yang Li, Xia Fan, Shuangyang Ding
Analytica Chimica Acta 2013 777() pp: 32-40
Publication Date(Web):
DOI:10.1016/j.aca.2013.03.026
Co-reporter:Zhiming Xiao, Xiaowei Li, Xiaolin Wang, Jianzhong Shen, Shuangyang Ding
Journal of Chromatography B 2011 Volume 879(Issue 1) pp:117-122
Publication Date(Web):1 January 2011
DOI:10.1016/j.jchromb.2010.11.008
A rapid, sensitive, and environmental-friendly method has been developed for the simultaneous determination of seven neonicotinoid insecticides residues in bovine muscle and liver. The sample preparation procedure was based on a high automated pressurized solvent extraction (PSE) combined with solid-phase extraction (SPE) clean-up. The target compounds were identified and quantitatively determined by liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC–ESI-MS/MS) operated in multiple reaction monitoring mode. Average recoveries of the seven analytes from fortified samples ranged between 83.2% and 101.9%, with relative standard deviations (RSDs) lower than 10.8%. The limits of detection (LODs) and quantification (LOQs) for neonicotinoids were in the ranges of 0.8–1.5 μg kg−1 and 2.5–5.0 μg kg−1, respectively. This validated method was successively applied to the determination of neonicotinoid insecticides in real samples from markets.
Co-reporter:Qiushi Huang, Jiancheng Li, Lijun Xia, Xi Xia, Peng Duan, Jianzhong Shen, Shuangyang Ding
Analytica Chimica Acta 2010 Volume 664(Issue 1) pp:62-67
Publication Date(Web):1 April 2010
DOI:10.1016/j.aca.2010.01.058
The depletion profile of valnemulin (VLM) was studied in healthy piglets after oral administration of a premix. Thirty pigs were given doses of 7.5 mg/kg body weight/day in the feed for 21 days. One control and five medicated piglets were randomly selected for sacrifice at 0.25, 0.5, 1, 2 and 3 days post-treatment. The residue concentrations of VLM in swine muscle, liver and kidney were detected using high performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS). The highest residue concentrations of VLM were attained in 0.25 day, and all of the samples were below the maximum residue limit (MRL) recommended by the European Medical Evaluation Agency (EMEA) at 0.5 day post-treatment. The residue concentrations of swine liver were significantly higher than those of kidney and muscle, which indicated liver to be the target tissue for VLM. The withdrawal period of VLM with oral administration was 24 hours.
Co-reporter:Xi Xia;Zhiming Xiao;Qiushi Huang;Lijun Xia;Kui Zhu;Xiaolin Wang
Chromatographia 2010 Volume 72( Issue 11-12) pp:1089-1095
Publication Date(Web):2010 December
DOI:10.1365/s10337-010-1799-z
A rapid and sensitive method has been developed for the simultaneous determination of four avermectins and one milbemycin residues in bovine tissue. The isolation of the analytes from muscle and liver samples was accomplished utilizing a pressurized solvent extractor. The optimized extraction procedure using acetonitrile/water (40:60, v/v) as extraction solvent, was automatically carried out at 100 °C and 10 MPa, applying two static cycles for 3 min. The extracts were cleaned up on a C18 solid-phase extraction cartridge and analyzed by liquid chromatography with fluorescence detection after derivatization. Mean recoveries of the five analytes from fortified samples were between 84.8 and 101.8%, with relative standard deviations lower than 10.8%. The limit of detection and quantification were in the ranges of 0.1–0.2 and 0.5–0.6 μg kg–1, respectively. The application of the newly developed method was demonstrated by analyzing bovine meat samples from market.
Co-reporter:Jianzhong Shen;Liming Guo;Fei Xu;Qinxiong Rao;Xi Xia;Xiaowei Li
Chromatographia 2010 Volume 71( Issue 5-6) pp:383-388
Publication Date(Web):2010 March
DOI:10.1365/s10337-009-1463-7
A rapid and sensitive analytical method for the simultaneous determination of four fluoroquinolones, four tetracyclines and six sulfonamides in chicken muscle using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC–MS–MS) has been developed and validated. Samples were extracted with McIlvaine buffer-acetonitrile, defatted with n-hexane, and analyzed by UPLC–MS–MS. Solvent delay technique was applied in the analysis to remove the non-volatile phosphate and carry out farther on-line SPE clean-up. Satisfactory recoveries (55–110%) of all the veterinary drugs were demonstrated in 1, 10 and 20 μg kg−1 spiked levels with the overall RSD for intra- and inter-day of 14 analytes less than 18%. The LOD and LOQ were 0.3 and 1.0 μg kg−1, respectively. Quantitative results of 103 real samples indicated that the present method was suitable for the quantitative analysis of real samples.
Co-reporter:Junxia Chen, Fei Xu, Haiyang Jiang, Yali Hou, Qinxiong Rao, Pengju Guo, Shuangyang Ding
Food Chemistry 2009 Volume 113(Issue 4) pp:1197-1201
Publication Date(Web):15 April 2009
DOI:10.1016/j.foodchem.2008.08.006
A rapid indirect competitive fluorescence-linked immunosorbent assay (cFLISA) based on quantum dots (QDs) as the fluorescent marker has been developed for the detection of Enrofloxacin (ENR) in chicken muscle tissue. The end-point fluorescent detection system was carried out using QDs conjugated with goat anti-mouse secondary antibody. The cFLISA method allowed for ENR determination in a liner working range of 1–100 ng mL−1 with the 50% inhibition value (IC50) of 8.3 ng mL−1 and the limit of detection (LOD) of 2.5 ng mL−1. The recoveries for chicken muscle samples spiked with ENR at levels of 50–200 μg kg−1 ranged from 81% to 94% with coefficients of variation (CV) of 10–13%. In real chicken tissue sample analysis, the results of cFLISA were similar to those obtained from an indirect competitive enzyme-linked immunosorbent assay (cELISA) to a high performance liquid chromatography method (HPLC), which indicated that cFLISA is suitable as screening method for the monitoring of veterinary drug residues.
Co-reporter:Jing Tong;Qinxiong Rao;Kui Zhu;Zhigang Jiang
Journal of Separation Science 2009 Volume 32( Issue 23-24) pp:4254-4260
Publication Date(Web):
DOI:10.1002/jssc.200900415
Abstract
This work demonstrates the potential of HPCE in the analysis of antibiotics in a complex matrix such as feedstuffs. Using 20 mM citric acid–40 mM Na2HPO4 buffer (pH 2.65), the five antibiotics, tetracycline, oxytetracycline, doxycycline, tilmicosin, and tylosin were successfully separated at 30 kV in a 64.5 cm×75 μm id capillary. Good repeatability, stability, and reliability of the method were supported by <10% CV with mean recoveries of >70%, and the limit of detection of the five analytes was 0.5–1 mg/kg. It was for the first time that a capillary electrophoretic method was employed to simultaneously detect five tetracycline and macrolide antibiotics in animal feeds.
Co-reporter:Jianzhong Shen, Xi Xia, Haiyang Jiang, Cun Li, Jiancheng Li, Xiaowei Li, Shuangyang Ding
Journal of Chromatography B 2009 Volume 877(14–15) pp:1523-1529
Publication Date(Web):15 May 2009
DOI:10.1016/j.jchromb.2009.03.040
A sensitive and reliable method using gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS) was developed for the simultaneous determination of chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF), and florfenicol amine (FFA) at trace levels in muscle and liver. Before extraction with ethyl acetate, CAP-d5 was added to tissue samples as internal standard. The organic extracts were frozen to remove lipid and further purified by liquid–liquid extraction (LLE) with hexane and solid-phase extraction (SPE) using Oasis HLB cartridges. The target compounds were derivatized with BSTFA + 1% TMCS prior to GC-NCI/MS determination in selected ion monitoring mode (SIM). The recovery values ranged from 78.5 to 105.5%, with relative standard deviations (RSD) <17%. The limits of detections (LODs) of 0.1 μg/kg for CAP and 0.5 μg/kg for TAP, FF, and FFA were obtain. Incurred sample and samples from local market were successfully analyzed using this method.
Co-reporter:Haiyang Jiang;Degang Zhou;Haiyan Li;Fei Xu;Cun Li;Jianzhong Shen
Chromatographia 2008 Volume 68( Issue 3-4) pp:259-262
Publication Date(Web):2008 August
DOI:10.1365/s10337-008-0688-1
A simple multi-residue method was developed for the simultaneous determination of abamectin, ivermectin, doramectin, and eprinomectin in rabbit feces. Samples were extracted with acetonitrile, cleaned up with a C18 solid-phase extraction cartridge, and analysed by LC with fluorescence detection after derivatization. Abamectin, ivermectin, doramectin, and eprinomectin were detected at levels of 2–500 ng g−1; the average recoveries ranged from 73.2 to 99.6% with relative standard deviations of 2.5–11.3%. The limits of detection were 0.1–0.4 ng g−1.
Co-reporter:Gali Bingga, Zhicheng Liu, Jianfeng Zhang, Yujun Zhu, Lifeng Lin, Shuangyang Ding, Pengju Guo
Molecular and Cellular Probes (October–December 2014) Volume 28(Issues 5–6) pp:271-278
Publication Date(Web):1 October 2014
DOI:10.1016/j.mcp.2014.08.001
A high resolution melting (HRM) curve method was developed to identify canine parvovirus type 2 (CPV-2) strains by nested PCR. Two sets of primers, CPV-426F/426R and CPV-87R/87F, were designed that amplified a 52 bp and 53 bp product from the viral VP2 capsid gene. The region amplified by CPV-426F/426R included the A4062G and T4064A mutations in CPV-2a, CPV-2b and CPV-2c. The region amplified by CPV-87F/87R included the A3045T mutation in the vaccine strains of CPV-2 and CPV-2a, CPV-2b and CPV-2c. Faecal samples were obtained from 30 dogs that were CPV antigen-positive. The DNA was isolated from the faecal samples and PCR-amplified using the two sets of primers, and genotyped by HRM curve analysis. The PCR-HRM assay was able to distinguish single nucleotide polymorphisms between CPV-2a, CPV-2b and CPV-2c using CPV-426F/426R. CPV-2a was distinguished from CPV-2b and CPV-2c by differences in the melting temperature. CPV-2b and CPV-2c could be distinguished based on the shape of the melting curve after generating heteroduplexes using a CPV-2b reference sample. The vaccine strains of CPV-2 were identified using CPV-87F/87R. Conventional methods for genotyping CPV strains are labor intensive, expensive or time consuming; the present PCR-based HRM assay might be an attractive alternative.
Co-reporter:Zhiming Xiao, Yunxia Yang, Yang Li, Xia Fan, Shuangyang Ding
Analytica Chimica Acta (13 May 2013) Volume 777() pp:32-40
Publication Date(Web):13 May 2013
DOI:10.1016/j.aca.2013.03.026
•A novel SWE technique for the determination of neonicotinoids in eels is presented.•Extraction parameters have been optimized along with the validation procedures.•The proposed method is accurate, sensitive, and environmental-friendly.•It was successfully applied for the analysis of neonicotinoids in real world samples.A rapid, sensitive, and environmental-friendly multi-residue method has been developed for the simultaneous determination of seven neonicotinoid insecticides (dinotefuran, nitenpyram, thiamethoxam, imidacloprid, clothianidin, acetamiprid, and thiacloprid) residues in eel samples. Subcritical water extraction was investigated as a novel and alternative technology for the extraction of neonicotinoids from eel matrices and the results were compared with the conventional ultrasonic and shaking extraction. The target compounds were identified and quantitatively determined by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC–MS/MS) operated in multiple reaction monitoring mode. Under the current optimized chromatographic conditions, each LC run was completed in 5 min. Average recoveries of the seven analytes from fortified samples ranged between 84.6% and 102.0%, with relative standard deviations (RSD) lower than 10.8%. The limits of detection (LODs) and quantification (LOQs) for neonicotinoids were in the ranges of 0.12–0.36 μg kg−1 and 0.42–1.12 μg kg−1, respectively. The proposed method is fast, sensitive, easy to perform, water-based thus more environmentally acceptable, making it applicable for high-throughput monitoring of insecticides residues in aquatic products.Download full-size image
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