Ex-treatment of immunoassay method for nodularin and cylindrospermopsins
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摘要: 采用固相萃取前处理技术和酶联免疫吸附(Enzyme-Linked ImmunoSorbent Assays, ELISAs)检测技术,研究了水样中节球藻和柱胞藻毒素的免疫前处理方法.由于两种藻毒素极性差异较大,分别对水样中节球藻和柱胞藻毒素采用不同的前处理方法,具体如下.①节球藻毒素:先后用10 mL甲醇和10 mL纯水活化C18柱,以3~4 mL/min的速度上样后用10 mL纯水淋洗,再用12 mL甲醇洗脱节球藻毒素,氮气吹干后纯水定容至1 mL待测.②柱胞藻毒素:先后用10 mL甲醇和10 mL纯水活化碳黑固相萃取柱,以2~3 mL/min的速度上样后用10 mL甲醇淋洗,再用12 mL含0.1%三氟乙酸的甲醇洗脱柱胞藻毒素,氮气吹干后用0.1%氨水定容至1 mL待测.结果表明,这两种前处理方法对节球藻与柱胞藻毒素的ELISAs检测能保证有较好的回收率, 加标回收率可分别达到99.4%和77%.Abstract: Effective solid phase extraction (SPE) ex-treatment method and sensitive enzyme-linked immunosorbent asssay (ELISA) were developed for determining the nodularins(NOD) and cylindrospermopsins(CYN) in water samples. Due to the different polarities, two different ex-treatment methods were applied to NOD and CYN, respectively. The methods were developed as followed: ① For NOD,firstly the C18 column was activated by 10 mL methanol and 10 mL Milli-Q water; then the sample was applied to the column at 3-4 mL/min; after washing the column with 10 mL water, the NOD was eluted with 10 mL methanol; then the eluent was dried up with a rotary evaporator, and lastly the final volume was fixed to 1 mL with Milli-Q water for ELISA analyse. ② For CYN, the carb SPE tubes were activated with 10mL methanol and 10mL Milli-Q water; then the sample was applied to the tubes at 2-3 mL/min; after washing the tubes with 10mL methanol, the CYN was eluted with 12 mL methanol containing 0.1% trifluoroacetic acid;then the eluent was dried up with a rotary evaporator, lastly the final volume was fixed to 1 mL with Milli-Q water containing 0.1% ammonia for ELISA analyse. The results indicated that these methods can guarantee the good recoveries of NOD and CYN for ELISA analyse, which could reach to 94.3% and 77%, respectively.
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Key words:
- nodularins /
- cylindrospermopsins /
- ELISAs
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