摘要
采用优化的碳二亚胺法制备诺氟沙星(NOR)人工抗原,免疫新西兰大白兔制备多克隆抗体,建立间接竞争ELISA标准曲线。对NOR的线性检测范围为0.07~31.8ng/mL,半数抑制质量浓度(IC50)和最低检测限(LOD)分别为1.5ng/mL和0.04ng/mL。检测缓冲液中pH值和磷酸根离子浓度的最佳参数分别为7.4和10mmol/L;对甲醇和NaOH溶液(30mmol/L)的最佳调整体积分数分别为30%和10%。抗体具有广谱特异性,与环丙沙星(83.3%)、培氟沙星(78.9%)、依诺沙星(68.2%)、恩诺沙星(51.7%)和洛美沙星(41.7%)有较高的交叉反应率,可用于氟喹诺酮类药物(FQs)多残留检测方法研究。
Modified EDC method was employed to synthesize norfloxacin(NOR) artificial antigen and New Zealand rabbits were immunized for anti-NOR polyclonal antibody(pAb) production.An indirect competitive ELISA standard curve was established.The developed assay was sensitive and had a linear range from 0.07 to 31.8 ng/mL(R2 = 0.9627),with IC50 and LOD of respectively 1.5 ng/mL and 0.04 ng/mL.The optimal pH and phosphate ion concentration in assay buffer were determined to be 7.4 mmol/L and 10 mmol/L,respectively,and the concentration adjustments for methanol and NaOH(30 mmol/L) in dilute solutions did not exceed 30% and 10%,respectively.The produced pAb had broad-spectrum specificity and exhibited a high cross-reactivity with ciprofloxacin(83.3%),pefloxacin(78.9%),enoxacin(68.2%),enrofloxacin(51.7%) and lomefloxacin(41.7%).The results obtained suggest that the established icELISA method be used for the multi-determination of fluoroquinolone residues.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2011年第21期148-151,共4页
Food Science
基金
河南省高校科技创新人才支持计划资助项目(2010HASTIT026)
关键词
诺氟沙星
氟喹诺酮
间接竞争ELISA
多残留检测
norfloxacin(NOR)
fluoroquinolone(FQs)
indirect competitive ELISA
multi-residue detection