Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics
A fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers (DMIPs) as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2016-05-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311916613576 |
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| author | Xin-wei DU Yan-xin ZHANG Yong-xin SHE Guang-yang LIU Feng-nian ZHAO Jing WANG Shan-shan WANG Fen JIN Hua SHAO Mao-jun JIN Lu-fei ZHENG |
| author_facet | Xin-wei DU Yan-xin ZHANG Yong-xin SHE Guang-yang LIU Feng-nian ZHAO Jing WANG Shan-shan WANG Fen JIN Hua SHAO Mao-jun JIN Lu-fei ZHENG |
| author_sort | Xin-wei DU |
| collection | DOAJ |
| description | A fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers (DMIPs) as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased binding of fluorescent substances to DMIPs due to increased concentrations of melamine in the solutions. DMIPs for melamine were synthesized under a hot water bath in the presence of the initiator azobisisobutyronitrile (AIBN) using 2,4-diamino-6-methyl-1,3,5-triazine (DAMT) as a dummy template, methacrylic acid (MAA) as a functional monomer, and ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent. The adsorption capacity and selectivity of DMIPs for melamine were evaluated by the isothermal adsorption curve and Scatchard analysis. The evaluation results showed that the synthesized DMIPs had specific recognition sites for melamine and the maximum adsorption amount was 1 066.33 μg g−1. Later, 5-(4,6-dichlorotriazinyl) amino fluorescein (DTAF) with a triazine ring, which slightly resembles melamine, was selected as the fluorescent substance. The fluorescent competitive assay using DMIPs as the antibody mimics was finally established by selecting and optimizing the reaction solvents, DMIPs amount, DTAF concentration, and incubation time. The optimal detection system showed a linear response within range of 0.05–40 mg L−1 and the limit of detection (LOD) was 1.23 μg L−1. It was successfully applied to the detection of melamine in spiked milk samples with satisfactory recoveries (71.9 to 86.3%). According to the comparative analysis, the result of optimized fluorescent competitive assay revealed excellent agreement with the HPLC-MS/MS result for melamine. |
| format | Article |
| id | doaj-art-f9122253178b43f8a3da1670c7cbcecb |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2016-05-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-f9122253178b43f8a3da1670c7cbcecb2025-08-20T03:57:55ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192016-05-011551166117710.1016/S2095-3119(16)61357-6Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimicsXin-wei DU0Yan-xin ZHANG1Yong-xin SHE2Guang-yang LIU3Feng-nian ZHAO4Jing WANG5Shan-shan WANG6Fen JIN7Hua SHAO8Mao-jun JIN9Lu-fei ZHENG10Institute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.China; DU Xin-wei, Mobile: +86-13581657461School of Chemical Engineering and Technology, Harbin Institute of Technology, Harbin 150001, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaSchool of Chemical Engineering and Technology, Harbin Institute of Technology, Harbin 150001, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.China; Correspondence WANG Jing, Tel: +86-10-82106568Institute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaInstitute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China; Key Laboratory of Agro-product Quality and Safety, Beijing 100081, P.R.ChinaA fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers (DMIPs) as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased binding of fluorescent substances to DMIPs due to increased concentrations of melamine in the solutions. DMIPs for melamine were synthesized under a hot water bath in the presence of the initiator azobisisobutyronitrile (AIBN) using 2,4-diamino-6-methyl-1,3,5-triazine (DAMT) as a dummy template, methacrylic acid (MAA) as a functional monomer, and ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent. The adsorption capacity and selectivity of DMIPs for melamine were evaluated by the isothermal adsorption curve and Scatchard analysis. The evaluation results showed that the synthesized DMIPs had specific recognition sites for melamine and the maximum adsorption amount was 1 066.33 μg g−1. Later, 5-(4,6-dichlorotriazinyl) amino fluorescein (DTAF) with a triazine ring, which slightly resembles melamine, was selected as the fluorescent substance. The fluorescent competitive assay using DMIPs as the antibody mimics was finally established by selecting and optimizing the reaction solvents, DMIPs amount, DTAF concentration, and incubation time. The optimal detection system showed a linear response within range of 0.05–40 mg L−1 and the limit of detection (LOD) was 1.23 μg L−1. It was successfully applied to the detection of melamine in spiked milk samples with satisfactory recoveries (71.9 to 86.3%). According to the comparative analysis, the result of optimized fluorescent competitive assay revealed excellent agreement with the HPLC-MS/MS result for melamine.http://www.sciencedirect.com/science/article/pii/S2095311916613576dummy molecularly imprinted polymersmelaminefluorescent competitive assayartificial antibody |
| spellingShingle | Xin-wei DU Yan-xin ZHANG Yong-xin SHE Guang-yang LIU Feng-nian ZHAO Jing WANG Shan-shan WANG Fen JIN Hua SHAO Mao-jun JIN Lu-fei ZHENG Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics Journal of Integrative Agriculture dummy molecularly imprinted polymers melamine fluorescent competitive assay artificial antibody |
| title | Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| title_full | Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| title_fullStr | Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| title_full_unstemmed | Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| title_short | Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| title_sort | fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics |
| topic | dummy molecularly imprinted polymers melamine fluorescent competitive assay artificial antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2095311916613576 |
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