摘要
采用碳酰二咪唑(CDI)将SepharoseCL-4B活化并与纯化的抗氯磺隆抗体共价偶联,合成免疫亲和吸附剂并制备对氯磺隆具有特异性亲和力的免疫亲合色谱(IAC)柱。对IAC条件进行优化,选择0.02mol.L-1pH7.2磷酸盐缓冲液作吸附与平衡介质,80%(体积分数)甲醇水作洗脱剂。结果表明,在实验条件下,IAC柱对氯磺隆的动态柱容量达2.6!g.mL-1床体积。当标样溶液中氯磺隆含量为2.0ng.mL-1时,经IAC柱富集后浓度提高近250倍。土壤中添加氯磺隆0.10!g.g-1、5.0ng.g-1,提取液以IAC柱分离富集,洗脱液采用高效液相色谱(HPLC)法测定,5次重复实验的平均回收率分别为92.9%、94.8%,相对标准偏差分别为8.46%、8.41%。成功建立了氯磺隆IAC-HPLC分析技术并用于土壤中痕量残留氯磺隆的测定。
The purified anti-chlorsulfuron antibody was conjugated to the CDI-activated Sepharose CL-4B to synthesize the immunosorbent and prepare the immunoaffinity chromatography （IAC） column for cleanup and concentration of chlorsulfuron in sample extract. The conditions of IAC were optimized, pH7.2 phosphate buffer was used as equilibrium and absorbent medium, 80% methanol was used as eluent. The results showed that the dynamic column capacity was up to 2.6 μg· mL^-1 bed volume under the experimental condition, the chlorsulfuron concentration was enhanced nearly 250 times by IAC when the initial concentration was 2.0 ng·mL^-1 in standard sample solution. The extract of spiked soil was cleaned up and concentrated by IAC, the chlorsulfuron in eluate was determined by high performance liquid chromatography （HPLC）, the average recoveries of chlorsulfuron was 92.9% and 94.8% with relative standard deviation of 8.46% and 8.41% at the spiked level of 0.10 μg· g^-1 and 5.0 ng·g^-1 respectively. The IAC-HPLC technology was developed successfully for the analysis of ehlorsulfuron in soil.
出处
《农业环境科学学报》
CAS
CSCD
北大核心
2006年第6期1663-1666,共4页
Journal of Agro-Environment Science
基金
国家自然科学基金资助项目(20177019)
