食品科学 ›› 2010, Vol. 31 ›› Issue (18): 350-354.doi: 10.7506/spkx1002-6630-201018083

• 分析检测 • 上一篇    下一篇

谷物中伏马菌素B1酶联免疫分析法的建立

刘师文,何庆华,邹 龙,许 杨*   

  1. 南昌大学 食品科学与技术国家重点实验室,中德联合研究院
  • 收稿日期:2010-05-07 修回日期:2010-08-27 出版日期:2010-09-25 发布日期:2010-12-29
  • 通讯作者: 许 杨* E-mail:xuyang1951@163.com
  • 基金资助:

    国家“863”计划项目(2007AA10Z427)

Development of an ELISA Method for Determining Fumonisin B1 in Cereals

LIU Shi-wen,HE Qing-hua,ZOU Long,XU Yang*   

  1. State Key Laboratory of Food Science and Technology, Sino-Germany Joint Research Institute, Nanchang University,
    Nanchang 330047, China
  • Received:2010-05-07 Revised:2010-08-27 Online:2010-09-25 Published:2010-12-29
  • Contact: XU Yang* E-mail:xuyang1951@163.com

摘要:

以匙孔血蓝蛋白(KLH)为载体,采用碳化亚二胺法人工合成伏马菌素B1(FB1)抗原FB1-KLH,免疫大白兔获得特异性良好的抗FB1 多克隆抗体;在多克隆抗体的基础上建立伏马菌素B1 的间接竞争酶联免疫吸附分析方法,该方法IC50 为11.7μg/L,最低检出限1.1μg/L,平均批内和批间变异系数分别为5.8%、10.2%;分析不同溶剂对标准曲线和加标回收率的影响,确定PBS 溶液为样本最佳提取溶剂;在玉米、小麦、大米、高粱谷物样本中添加50~500μg/kg 的FB1 标准品,平均回收率在70.5%~105.6% 之间。将该方法应用于40 份谷物样本中FB1 的检测,检测结果与高效液相色谱的相关系数(R2)为0.9547。该方法简单、灵敏、快速、准确,适用于谷物中FB1 的检测。

关键词: 伏马菌素B1, 酶联免疫分析(ELISA), 谷物

Abstract:

Polyclonal antibodies against fumonisin B1 were produced in rabbits with FB1/keyhole limpet hemocyanin conjugate. A sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) for determining FB1 was developed. The concentration of FB1 causing 50% inhibition of binding enzyme marker (IC50) was 11.7μg/L and the detection limit was 1.1μg/L. The average variation coefficients in intragroup and intergroup were 5.8% and 10.2%, respectively. PBS was selected as the best extract solvent through extensive investigations. The recovery rates from corn, wheat, rice and sorghum samples spiked at a level varying from 50 to 500μg/kg ranged from 70.5% to105.6%. The contents of FB1 in 40 cereal samples were screened by the developed ELISA method and the results exhibited a positive correlation with the results from HPLC (R2 = 0.9547). This method is sensitive, fast and accurate and therefore is suitable for screening the presence of FB1 in cereals.

Key words: fumonisin B1, ELISA, cereal

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