Spirotetramat, a new type of insecticide which can destroy the energy metabolism of pests by acting as an acetyl-CoA carboxylase inhibitor, is widely used in the prevention of sucking pests on vegetables and fruits. However, spirotetramat can cause eye impairment and skin sensitization for animals and human beings. The aim of this study was to achieve rapid detection of spirotetramat residues in vegetables, fruits and environmental samples by preparing monoclonal antibody of spirotetramat and establish an enzyme-linked immunoassay. The spirotetramat hapten was conjugated to BSA to prepare immunogen by the active ester method. One monoclonal antibody (mAb 3D11G7) against both spirotetramat and its metabolite B-enol was prepared. Under optimized conditions, an indirect homologous competitive enzyme-linked immunosorbent assay (icELISA) method was developed. The inhibitor median concentration (IC50) value was 2.1 μg/L and the working range was 0.5-8.6 μg/L. The recoveries of spirotetramat detected in water from Yangtze River, soil, tomato and citrus by icELISA method were ranged from 72.9% to 110.1%. The concentrations of spirotetramat in citrus samples detected by icELISA was in accordance with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) (R2=0.965 2). All these results showed that the developed icELISA method can be used to monitor spirotetramat residue in vegetables, fruits and environmental samples.
ZHAO Jing
,
ZHAO Qiyang
,
ZHANG Yaohai
,
HE Yue
,
JIAO Bining
,
CUI Yongliang
. Preparation of monoclonal antibody-based immunoassay for the detection of spirotetramat[J]. Food and Fermentation Industries, 2022
, 48(10)
: 247
-254
.
DOI: 10.13995/j.cnki.11-1802/ts.028552
[1] TIAN F J, QIAO C K, WANG C X, et al.Simultaneous determination of spirodiclofen, spiromesifen, and spirotetramat and their relevant metabolites in edible fungi using ultra-performance liquid chromatography/tandem mass spectrometry[J].Scientific Reports, 2021, 11(1):1 547.
[2] CHEN X J, MENG Z Y, ZHANG Y Y, et al.Degradation kinetics and pathways of spirotetramat in different parts of spinach plant and in the soil[J].Environmental Science and Pollution Research International, 2016, 23(15):15 053-15 062.
[3] 叶玉凤. 螺虫乙酯在柑橘和土壤中的残留规律和降解动态研究[D].重庆:西南大学, 2018.
YE Y F.Study on residues rule and degradation dynamics of spirotetramat in citrus and soil[D].Chongqing:Southwest University, 2018.
[4] ZHANG Q M, ZHANG G L, YIN P J, et al.Toxicological effects of soil contaminated with spirotetramat to the earthworm Eisenia fetida[J].Chemosphere, 2015, 139:138-145.
[5] WU H M, RAO Q, ZHENG J Y, et al.Biochemical and histological alterations in adult zebrafish (Danio rerio) ovary following exposure to the tetronic acid insecticide spirotetramat[J].Ecotoxicology and Environmental Safety, 2018, 164:149-154.
[6] ZHANG J, QIAN L, WANG C, et al.Dysregulation of endocrine disruption, apoptosis and the transgenerational toxicity induced by spirotetramat[J].Chemosphere, 2020, 240:124900.
[7] YIN X H, JIANG S J, YU J, et al.Effects of spirotetramat on the acute toxicity, oxidative stress, and lipid peroxidation in Chinese toad (Bufo bufo gargarizans) tadpoles[J].Environmental Toxicology and Pharmacology, 2014, 37(3):1 229-1 235.
[8] ANASTASSIADOU M, BELLISAI G, BERNASCONI G, et al.Modification of the existing maximum residue levels for spirotetramat in leeks, spring onions and honey[J].EFSA Journal European Food Safety Authority, 2021, 19(3):6 517.
[9] 张蕊, 杨石有, 张贺, 等.高效液相色谱法测定螺虫乙酯在黄瓜幼苗中的内吸传导特性[J].农药, 2019, 58(2):99-102.
ZHANG R, YANG S Y, ZHANG H, et al.The intake transportation function of spirotetramat in cucumber using high performance liquid chromatography[J].Agrochemicals, 2019, 58(2):99-102.
[10] ZHANG J, QIAN L, WANG C, et al.UPLC-TOF-MS/MS metabolomics analysis of zebrafish metabolism by spirotetramat[J].Environmental Pollution, 2020, 266:115310.
[11] 杨松, 王瑶, 禾丽菲, 等.QuEChERS-气相色谱-串联质谱法分析鲜茶叶中6种杀虫剂的残留及消解动态[J].分析试验室, 2019, 38(12):1 459-1 464.
YANG S, WANG Y, HE L F, et al.Residue and dissipation dynamics of six insecticides in fresh tea by QuEChERS-gas chromatography tandem mass spectrometry[J].Chinese Journal of Analysis Laboratory, 2019, 38(12):1 459-1 464.
[12] LAN J Q, ZHAO H W, JIN X T, et al.Development of a monoclonal antibody-based immunoaffinity chromatography and a sensitive immunoassay for detection of spinosyn A in milk, fruits, and vegetables[J].Food Control, 2019, 95:196-205.
[13] CEVALLOS-CEDEÑO R E, AGULLÓ C, ABAD-SOMOVILLA A, et al.Hapten design and antibody generation for immunoanalysis of spirotetramat and spirotetramat-enol[J].ACS Omega, 2018, 3(9):11 950-11 957.
[14] CEVALLOS-CEDEÑO R E, AGULLO C, ABAD-FUENTES A, et al.Enzyme and lateral flow monoclonal antibody-based immunoassays to simultaneously determine spirotetramat and spirotetramat-enol in foodstuffs[J].Scientific Reports, 2021, 11(1):1 809.
[15] CUI Y L, ZHAO J, ZHOU J, et al.Development of a sensitive monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for analysing nobiletin in citrus and herb samples[J].Food Chemistry, 2019, 293:144-150.
[16] 郭素琴. 青蒿素类药物特异性单克隆抗体的制备及快速免疫检测方法的建立与应用[D].北京:中国农业大学, 2016.
GUO S Q.Development and application of specific monoclonal antibodies and immunoassay techniques for artemisinin and its derivatives[D].Beijing:China Agricultural University, 2016.
[17] ZHAO J, ZHANG Y H, ZHAO Q Y, et al.A sensitive and practical ELISA for analyzing naringenin in pummelo and herb samples[J].Food Chemistry, 2021, 362:130223.
[18] 李壮, 张维库, 张越, 等.梓醇人工抗原的合成、鉴定及免疫原性的初步研究[J].中国中药杂志, 2015, 40(7):1 287-1 290.
LI Z, ZHANG W K, ZHANG Y, et al.Synthesis, identification of artificial antigen of catalpol and preliminary study of immunogenicity[J].China Journal of Chinese Materia Medica, 2015, 40(7):1 287-1 290.
[19] ZHANG B, NAN T G, ZHAN Z L, et al.A monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of chlorogenic acid in honeysuckle[J].Journal of Pharmaceutical and Biomedical Analysis, 2018, 148:1-5.
[20] LIU Y Y, SU X S, JIAN Q, et al.Behaviour of spirotetramat residues and its four metabolites in citrus marmalade during home processing[J].Food Additives and Contaminants Part A:Chemistry, Analysis, Control, Exposure & Risk Assessment, 2016, 33(3):452-459.
