节球藻和柱胞藻毒素免疫分析的前处理方法研究

华东师范大学学报(自然科学版) ›› 2011, Vol. 2011 ›› Issue (6): 108-114.

节球藻和柱胞藻毒素免疫分析的前处理方法研究

李双¹，殷浩文²

1.华东师范大学生命科学学院，上海 200062； 2.上海市检测中心，上海 201203

收稿日期:2010-11-01 修回日期:2011-02-01 出版日期:2011-11-25 发布日期:2011-11-22

Ex-treatment of immunoassay method for nodularin and cylindrospermopsins

LI Shuang¹, YIN Hao-wen²

1. School of Life Science, East China Normal University, Shanghai 200062, China;
2. The Bioassay and Safety Assessment,Shanghai Academy of Public Measurement, Shanghai 201203, China

Received:2010-11-01 Revised:2011-02-01 Online:2011-11-25 Published:2011-11-22

摘要/Abstract

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%.

关键词: 节球藻毒素, 柱胞藻毒素, ELISAs

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.

Key words: nodularins, cylindrospermopsins, ELISAs

中图分类号:

李双，殷浩文. 节球藻和柱胞藻毒素免疫分析的前处理方法研究[J]. 华东师范大学学报(自然科学版), 2011, 2011(6): 108-114.

LI Shuang, YIN Hao-wen. Ex-treatment of immunoassay method for nodularin and cylindrospermopsins[J]. Journal of East China Normal University(Natural Sc, 2011, 2011(6): 108-114.

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