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用于测定油炸食品中丙烯酰胺的高通量酶联免疫吸附测定法的分析评估。

Analytical evaluation of a high-throughput enzyme-linked immunosorbent assay for acrylamide determination in fried foods.

作者信息

Franek Milan, Rubio Daniel, Diblikova Iva, Rubio Fernando

机构信息

Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.

Abraxis LLC, 54 Steamwhistle Drive, Warminster, PA 18974, USA.

出版信息

Talanta. 2014 Jun;123:146-50. doi: 10.1016/j.talanta.2014.02.007. Epub 2014 Feb 15.

Abstract

The analytical performance and evaluation of a kit-based ELISA for the determination of acrylamide in fried potato and corn chip samples are described. The sample homogenate is subjected to clean-up using SPE, followed by analyte derivatization and ELISA detection. Accuracy, precision and linearity of the ELISA procedure have been validated using spiked samples. Analytical recovery ranged from 91.8% to 96.0% with coefficients of variation below 15%. Good linearity over a wide range of dilution and minimal assay drift was observed within a microtiter plate. IC50 value of the calibration curve was 110 ng/mL, with the limit of detection about 5 ng/mL and dynamic range from 10 to 1000 ng/mL. The high specificity of the ELISA was demonstrated by cross-reactivity study using 11 potential cross-reactants. A good correlation between the results obtained from the ELISA and GC-MS within the concentration range 120-1500 μg/kg was found in the chip samples (r=0.992, n=120). The data demonstrate that the evaluated and validated ELISA has a potential utility in a quick, simple and reliable acrylamide screening analysis for the medium- and large-sized food companies, as well as for residue laboratories and the food industry dealing with improving the chemical safety of foods available to the consumer.

摘要

本文描述了一种基于试剂盒的酶联免疫吸附测定法(ELISA)用于测定炸薯条和玉米片样品中丙烯酰胺的分析性能及评估。样品匀浆经固相萃取(SPE)净化,随后进行分析物衍生化及ELISA检测。通过加标样品验证了ELISA方法的准确度、精密度和线性。分析回收率在91.8%至96.0%之间,变异系数低于15%。在微量滴定板内观察到在较宽稀释范围内具有良好线性且测定漂移极小。校准曲线的半数抑制浓度(IC50)值为110 ng/mL,检测限约为5 ng/mL,动态范围为10至1000 ng/mL。使用11种潜在交叉反应物进行交叉反应性研究,证明了ELISA具有高特异性。在芯片样品中,浓度范围为120 - 1500 μg/kg时,ELISA与气相色谱 - 质谱联用仪(GC - MS)所得结果之间存在良好相关性(r = 0.992,n = 120)。数据表明,经评估和验证的ELISA在为中型和大型食品公司以及残留实验室和致力于提高消费者可获得食品化学安全性的食品行业进行快速、简单且可靠的丙烯酰胺筛查分析方面具有潜在用途。

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