Abstract：
Methyl parathion, a highly toxic, efficient, and persistent organophosphorus pesticide, is widely used in China. Sibutramine, a non-amphetamine central nervous system depressant, helps lose weight by disrupting hormone regulation, stimulating sympathetic nerves, and suppressing appetite. However, some unethical businesses fail to properly handle raw materials in foods like apple cider vinegar, leading to residual methyl parathion in apples or illegal excessive addition of sibutramine. Therefore, it is imperative to develop an immunoassay for the rapid detection of methyl parathion and sibutramine. The corresponding two haptens were prepared and coupled with the carrier proteins according to methyl parathion-sulfur-bovine serum protein (BSA)/chicken ovalbumin (OVA)-sibutramine (20 : 1 : excess, 15 : 1 : excess, 10 : 1 : excess, and 5 : 1 : excess), and sibutramine-BSA/OVA-methyl parathion (20 : 1 : excess, 10 : 1 : excess: 5 : 1 : excess, and 0 : 1 : excess). The result shows that the inhibition rate of the antibody obtained by methyl parathion-BSA/OVA-sibutramine (20 : 1 : excess) was higher than that of sibutramine-BSA/OVA-methyl parathion, which was 67.93%, and the concentration of methyl parathion was 8.65 ng mL-1 at this inhibition rate. Thus, methyl parathion-BSA/OVA-sibutramine (8.65 : 1 : excess) and the corresponding antibodies were selected for subsequent method establishment. By changing the concentration of the coating and antibody, the inhibition rate was found when the coating was 0.125 ng mL-1 and the antibody was diluted 4000 times. The antibody was used to develop a standard curve for the detection of sibutramine at the half-maximum inhibitory concentration (IC50) is 4.59 ng mL-1, the limit of detection (IC10) is 2.21 ng mL-1, the detection range is 2.89 to 7.28 ng mL-1, methyl p-phosphorus at the half-maximum inhibitory concentration (IC50) is 15.34 ng mL-1, the limit of detection (IC10) is 0.42 ng mL-1, the detection range is ng mL-1. Under these conditions, the recovery rate was between 88% and 102%, within reasonable limits, indicating the successful establishment of a rapid enzyme-linked ELISA assay.
摘要：
甲基对硫磷,剧毒,高效,和持久性有机磷农药，在中国广泛使用。西布曲明,一种非苯丙胺中枢神经系统抑制剂，通过破坏激素调节来帮助减肥，刺激交感神经，抑制食欲。然而,一些不道德的企业未能正确处理苹果醋等食品中的原材料，导致苹果中残留甲基对硫磷或非法过量添加西布曲明。因此,必须开发一种快速检测甲基对硫磷和西布曲明的免疫分析方法。根据甲基对硫磷-硫-牛血清蛋白（BSA）/鸡卵清蛋白（OVA）-西布曲明（20：1：过量，15:1:超额,10:1:超额,和5：1：超额），和西布曲明-BSA/OVA-甲基对硫磷（20：1：过量，10:1:超额:5:1:超额,和0：1：超额）。结果表明,甲基对硫磷-BSA/OVA-西布曲明(20:1:过量)抗体的抑制率高于西布曲明-BSA/OVA-甲基对硫磷,67.93%，在此抑制率下，甲基对硫磷的浓度为8.65ngmL-1。因此，选择甲基对硫磷-BSA/OVA-西布曲明(8.65:1:过量)和相应的抗体用于随后的方法建立。通过改变涂层和抗体的浓度，当涂层为0.125ngmL-1，抗体稀释4000倍时，发现抑制率。该抗体用于建立标准曲线，用于检测西布曲明的半数最大抑制浓度（IC50）为4.59ngmL-1，检测限（IC10）为2.21ngmL-1，检测范围为2.89至7.28ngmL-1，甲基对磷的半数最大抑制浓度（IC50）为15.34ngmL-1，检测限（IC10）为0.42ng-1mL。在这些条件下,回收率在88%到102%之间,在合理的范围内,表明成功建立了快速酶联ELISA检测方法。
