Immunochromatography of fusarochromanone mycotoxins.

J H Yu, F S Chu
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Abstract

The present paper describes an enzyme-linked immunoassay (ELISA) used in combination with thin-layer chromatography (TLC) and liquid chromatography (LC) for determination of fusarochromanone (TDP) mycotoxins in barley, wheat, and a Fusarium culture grown in rice and corn. The mycotoxins were first extracted from the sample with 100% methanol and subjected to TLC or LC without additional cleanup treatment. Individual fractions eluted from TLC or LC were acetylated, then analyzed by ELISA. Determinations of TDP toxins at levels as low as 0.1 and 0.5 ng were achieved by ELISA in combination with LC and TLC, respectively. The detection limit for TDP-1 in barley and wheat was about 20 ppb by ELISA alone as compared with a detection limit of 5 ppb by a combination of ELISA with either TLC or LC. Overall analytical recovery (% of added) of TDP-1 added to barley and wheat at 5, 10, and 20 ppb of TDP-1 was 106.9 +/- 15.3 and 113.2 +/- 11.6 by LC-ELISA and 108.8 +/- 9.1 and 110.4 +/- 4.9 by TLC-ELISA, respectively. Analysis of extracts obtained from Fusarium equiseti R6137 grown in corn and rice by the combination of TLC and ELISA revealed that diacetyl-TDP was also produced by this fungus in addition to TDP-1 and TDP-2. Comparable results were obtained when fungal extracts were subjected to ELISA, LC, and immunochromatography (i.e., combination of ELISA with either TLC or LC).

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褐黄酮真菌毒素的免疫层析。
本文介绍了用酶联免疫分析法(ELISA)联合薄层色谱法(TLC)和液相色谱法(LC)测定大麦、小麦、水稻和玉米中镰刀菌素(fusarochromanone, TDP)真菌毒素的方法。首先用100%甲醇从样品中提取真菌毒素,并进行薄层色谱或LC,不进行额外的清理处理。从TLC或LC洗脱的单个组分乙酰化,然后用ELISA分析。ELISA联合LC和TLC分别检测了低至0.1和0.5 ng的TDP毒素。ELISA对大麦和小麦中TDP-1的检出限为20 ppb,而ELISA与TLC或LC联合检测的检出限为5 ppb。在5、10和20 ppb的TDP-1浓度下,大麦和小麦中TDP-1的总分析回收率(添加量的百分比)分别为106.9 +/- 15.3和113.2 +/- 11.6,TLC-ELISA法为108.8 +/- 9.1和110.4 +/- 4.9。对玉米和水稻中生长的镰刀菌R6137提取液进行薄层色谱和酶联免疫吸附分析,发现该真菌除产生TDP-1和TDP-2外,还产生二乙酰基tdp。当真菌提取物进行ELISA、LC和免疫层析(即ELISA与TLC或LC的组合)时,获得了类似的结果。
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