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首页> 外文期刊>Brazilian Archives of Biology and Technology >A Comparison between Enzyme Immunoassay and HPLC for Ochratoxin A Detection in Green, Roasted and Instant Coffee
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A Comparison between Enzyme Immunoassay and HPLC for Ochratoxin A Detection in Green, Roasted and Instant Coffee

机译：酶免疫法与高效液相色谱法检测生咖啡，烘焙咖啡和速溶咖啡中O曲毒素A的比较

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摘要

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green,roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents preparedwere OTA-BSA (4.76 mu g/mL), anti-OTA.7 MAb (2xl0~3-fold dilution) and HRP-anti IgG (10~3-fold dilution). Thedetection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performanceliquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic-ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee,46.73 %for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %,respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. Theresults indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection,contributing to quality and safety of coffee products.
机译：使用抗OTA单克隆抗体开发了一种间接竞争酶联免疫吸附测定法（ic-ELISA），用于绿色，烘焙和速溶咖啡中的ra曲霉毒素A（OTA）检测。制备的免疫试剂为OTA-BSA（4.76μg/ mL），抗-OTA.7 MAb（2x10〜3倍稀释）和HRP-抗IgG（10〜3倍稀释）。检出限为3.73 ng OTA / g，该免疫测定与高效液相色谱之间的相关系数（r）：生咖啡为0.98，焙炒为0.98，速溶咖啡为0.86。通过ic-ELISA检测到的OTA水平高于HPLC，其中ELISA / HPLC比为0.66-1.46（生咖啡），0.96-1.11（烘焙）和0.93-1.82（即食）。添加到咖啡中（5-70 ng / g）的OTA的ELISA回收率：生咖啡为81.53％，焙炒为46.73％，速溶咖啡为64.35％，而HPLC的回收率分别为80.54％，45.91％和55.15％。在进行ELISA分析之前，通过样品稀释将基质干扰降至最低。结果表明，基于单克隆抗体的ic-ELISA可能是一种简单，灵敏，特异的OTA检测筛选工具，有助于提高咖啡产品的质量和安全性。

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