农业环境科学学报
農業環境科學學報
농업배경과학학보
Journal of Agro-Environment Science
2009年
10期
2211-2216
,共6页
韦林洪%刘曙照%邵秀金%张军
韋林洪%劉曙照%邵秀金%張軍
위림홍%류서조%소수금%장군
绿磺隆%克百威%三唑磷%多抗体免疫亲和色谱(MIAC)
綠磺隆%剋百威%三唑燐%多抗體免疫親和色譜(MIAC)
록광륭%극백위%삼서린%다항체면역친화색보(MIAC)
chlorsulfuron%carbofuran%triazophos%multi-class immunoaffinity chromatography (MIAC)
采用碳酰二咪唑(CDI)将Sepharose CL-4B活化并分别与纯化的抗绿磺隆、抗克百威、抗三唑磷的多克隆抗体共价偶联,合成相应的免疫亲和吸附剂,并制备了对绿磺隆、克百威和三唑磷具有特异性亲和力的多抗体免疫亲和色谱(MIAC)柱.对MIAC的条件进行优化,选择0.02 mol·L~(-1) pH7.2磷酸盐缓冲液作吸附与平衡介质,80%(体积分数)甲醇水溶液作洗脱剂.结果表明,在上述试验条件下,MIAC柱对绿磺隆、克百威和I唑磷的动态柱容量分别达1.81、2.29和1.89μg·mL~(-1)床体积.用MIAC柱对添加有绿磺隆、克百威、三唑磷的河水与土壤提取液进行分离富集,洗脱液分别采用包被抗体直接竞争酶联免疫吸附(ELISA)法和高效液相色谱(HPLC)法测定,重复5次,平均回收率是89.98%～106.2%,相对标准偏差(RSD)为3.21%～14.81%,ELISA法和HPLC法的测定结果基本一致.成功建立了绿磺隆、克百威和三唑磷的多抗体免疫亲和色谱分析技术并用于河水和土壤中绿磺隆、克百威、三唑磷残留的测定.
採用碳酰二咪唑(CDI)將Sepharose CL-4B活化併分彆與純化的抗綠磺隆、抗剋百威、抗三唑燐的多剋隆抗體共價偶聯,閤成相應的免疫親和吸附劑,併製備瞭對綠磺隆、剋百威和三唑燐具有特異性親和力的多抗體免疫親和色譜(MIAC)柱.對MIAC的條件進行優化,選擇0.02 mol·L~(-1) pH7.2燐痠鹽緩遲液作吸附與平衡介質,80%(體積分數)甲醇水溶液作洗脫劑.結果錶明,在上述試驗條件下,MIAC柱對綠磺隆、剋百威和I唑燐的動態柱容量分彆達1.81、2.29和1.89μg·mL~(-1)床體積.用MIAC柱對添加有綠磺隆、剋百威、三唑燐的河水與土壤提取液進行分離富集,洗脫液分彆採用包被抗體直接競爭酶聯免疫吸附(ELISA)法和高效液相色譜(HPLC)法測定,重複5次,平均迴收率是89.98%～106.2%,相對標準偏差(RSD)為3.21%～14.81%,ELISA法和HPLC法的測定結果基本一緻.成功建立瞭綠磺隆、剋百威和三唑燐的多抗體免疫親和色譜分析技術併用于河水和土壤中綠磺隆、剋百威、三唑燐殘留的測定.
채용탄선이미서(CDI)장Sepharose CL-4B활화병분별여순화적항록광륭、항극백위、항삼서린적다극륭항체공개우련,합성상응적면역친화흡부제,병제비료대록광륭、극백위화삼서린구유특이성친화력적다항체면역친화색보(MIAC)주.대MIAC적조건진행우화,선택0.02 mol·L~(-1) pH7.2린산염완충액작흡부여평형개질,80%(체적분수)갑순수용액작세탈제.결과표명,재상술시험조건하,MIAC주대록광륭、극백위화I서린적동태주용량분별체1.81、2.29화1.89μg·mL~(-1)상체적.용MIAC주대첨가유록광륭、극백위、삼서린적하수여토양제취액진행분리부집,세탈액분별채용포피항체직접경쟁매련면역흡부(ELISA)법화고효액상색보(HPLC)법측정,중복5차,평균회수솔시89.98%～106.2%,상대표준편차(RSD)위3.21%～14.81%,ELISA법화HPLC법적측정결과기본일치.성공건립료록광륭、극백위화삼서린적다항체면역친화색보분석기술병용우하수화토양중록광륭、극백위、삼서린잔류적측정.
The purified anti-chlorsulfuron antibody, anti- carbofuran antibody and anti-triazophos antibody were conjugated to the CDI-acti-vated sepharose CL-4B respectively to synthesize the immunosorbent. The mixed immunosorbent of chlorsulfuron, carbofuran and triaophos were used to prepare multi-class immunoaffinity chromatography( MIAC) column which are specific to chlorsulfuron, carbofuran and triaophos. The conditions of MIAC were optimized, 0.02 mol·L~(-1) pH 7.2 phosphate buffer was used as equilibrium and absorbent medium, and 80% methanol-water( V/V ) as eluent. Under the experimental conditions, the dynamic column capacity of chlorsulfuron, carbofuran and triaophos were up to 1.81, 2.29 and 1.89 μg·mL~(-1) bed volume respectively. The spiked river water and the extracts of spiked soil were clean up and en-riched by MIAC,the target compounds in eluates were determined by enzyme linked immunosorbent assay(ELISA) and verified by high per-formance liquid chromatography(HPLC). The average recoveries were 89.98%～106.2% and the RSD were 3.21%～14.81%(n=5). The results from ELISA were well corresponded with those from HPLC. MIAC technology was developed successfully for analysis of chlorsulfuron, carbo-furan and triaophos residues in water river and soil.
