title: Quantitative determination of major capsaicinoids in serum by ELISA and time-resolved fluorescent immunoassay based on monoclonal antibodies.
abstract: To monitor capsaicinoids in serum on-site, three new monoclonal antibodies (mAbs) were firstly proposed using a conjugate of 4-[(4-hydroxy-3-methoxybenzyl) amino]-4-oxobutanoic acid as the immunogen. Among them, the YQQD8 mAb showed the highest sensitivity and cross-reactivity to major capsaicinoids, such as capsaicin, dihydrocapsaicin and N-vanillylnonanamide. A competitive indirect enzyme-linked immunosorbent assay (icELISA) and a time-resolved fluorescent immunochromatographic assay (TRFICA) were established based on this mAb. The linear range was 1.1-27.0ngmL(-1) for icELISA and 1.9-62.5ngmL(-1) for TRFICA and the limit of detection (LOD) of TRFICA was 1.5ngmL(-1). To decrease the interference of sample components and increase accuracy, serum samples were diluted four times before assays. As a result, the linear range of serum samples was 4.6-107.9ngmL(-1) for icELISA and 7.6-250.0ngmL(-1) for TRFICA. Both icELISA and TRFICA showed good recoveries (91.0-112.8% for icELISA and 87.6-111.5% for TRFICA) and concordant results in spiked experiments. Overall, this is the first report of immunoassay based on the mAbs for quantitative determination of major capsaicinoids, and the results demonstrate that both methods can meet the demands of rapid on-site assay for capsaicinoids in serum samples.
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Fei
authors.initials: F
authors.lastname: Ma
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, PR China. Electronic address: peiwuli@oilcrops.cn.
authors.forename: Peiwu
authors.initials: P
authors.lastname: Li
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Liangxiao
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Wen
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, PR China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Ding
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products, Wuhan, Ministry of Agriculture, Wuhan 430062, PR China. Electronic address: zhangqi521x@126.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D002211:Capsaicin / Q000031:analogs & derivatives* / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005453:Fluorescence; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D008807:Mice, Inbred BALB C; D011817:Rabbits; D051381:Rats; D014641:Vanillic Acid / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; C005871:vanillyl-N-nonylamide; D014641:Vanillic Acid; D002211:Capsaicin; C012906:dihydrocapsaicin
keywords: Capsaicin; Capsaicinoids; Indirect competitive enzyme-linked immunosorbent assay (icELISA); Monoclonal antibody; Time-resolved fluorescent immunochromatographic assay (TRFICA)