Zhou Y X, Yan Q J, Ci Y X, Guo Z Q, Rong K T, Chang W B, Zhao Y F
Department of Chemistry, Peking University, Beijing, China.
Arch Toxicol. 1995;69(9):644-8. doi: 10.1007/s002040050226.
Two artificial antigens, NalphaNepsilon-di(O,O-diisopropyl) phosphoryl L-lysine (DIP)- bovine serum albumin (BSA) conjugate (DIP-BSA) and DIP-KLH (keyhole limpet hemocyanin), were synthesized. Antibodies against sarin (O-isopropyl methylphosphonofluoridate) were obtained after immunization of rabbits with DIP-KLH conjugate. A competitive inhibition enzyme immunoassay (CIEIA) was developed to detect the organophosphorus nerve agent sarin. The antibody solutions could be inhibited by sarin as low as 10(-6) mol/l, and the standard curve was linear over 3 orders of magnitude. The coefficients of intraassay and interassay variation of this method were 5.4-6.2% (n = 11) and 8.0-9.5% (n = 6) at a sarin concentration range of 10(-3)-10(-6) mol/l, respectively. The recovery of sarin in water samples at the concentration of 5 x 10(-5) mol/l was in the range of 96.8-102.5%. The specificity of the antiserum was assessed by comparing the inhibition induced by sarin with soman, Vx, isopropyl alcohol and isopropyl methyl phosphonic acid. The results showed that less than 5 mmol/l soman, 2 mmol/l Vx, 16 mmol/l isopropyl alcohol and 8 mmol/l isopropyl methyl phosphonic acid did not influence the determination of sarin in water samples.
合成了两种人工抗原,即NαNε-二(O,O-二异丙基)磷酰-L-赖氨酸(DIP)-牛血清白蛋白(BSA)偶联物(DIP-BSA)和DIP-血蓝蛋白(KLH)。用DIP-KLH偶联物免疫家兔后获得了抗沙林(O-异丙基甲基膦酰氟)抗体。建立了一种竞争性抑制酶免疫分析法(CIEIA)来检测有机磷神经毒剂沙林。沙林对抗体溶液的抑制浓度低至10^(-6) mol/L,标准曲线在3个数量级内呈线性。在沙林浓度范围为10^(-3)-10^(-6) mol/L时,该方法的批内变异系数和批间变异系数分别为5.4-6.2%(n = 11)和8.0-9.5%(n = 6)。5×10^(-5) mol/L浓度水样中沙林的回收率在96.8-102.5%范围内。通过比较沙林与梭曼、维埃克斯、异丙醇和异丙基甲基膦酸诱导的抑制作用来评估抗血清的特异性。结果表明,低于5 mmol/L的梭曼、2 mmol/L的维埃克斯、16 mmol/L的异丙醇和8 mmol/L的异丙基甲基膦酸不影响水样中沙林的测定。