免疫亲和高效液相色谱法测定小麦粉中脱氧雪腐镰刀菌烯醇的方法改进

Improved Method for Determination of Deoxynivalenol in Wheat Flour by Immunoaffinity-High Performance Liquid Chromatography

摘要:

本文研究了小麦粉中脱氧雪腐镰刀菌烯醇的高效液相色谱的检测方法。小麦粉中的脱氧雪腐镰刀菌烯醇的检测对于小麦粉的储藏及食用质量安全控制非常重要，而GB 5009.111-2016中规定的免疫亲和高效液相色谱法的色谱条件对于检测小麦粉中脱氧雪腐镰刀菌烯醇存在假阳性的风险。本文在国家标准的基础上，通过试验研究，优化了液相色谱条件：流动相为甲醇-水（25:75，V/V），检测波长240 nm、扫描范围190 nm~400 nm，本文还通过二极管阵列检测器能进行全波长扫描的特性，研究了定性分析的方法。该方法验证结果：回收率为91.7%~97.2%，检出限为20 μg/kg，在50 ng/mL~5000 ng/mL的浓度范围内，线性相关系数为0.99989，精密度为0.83%~3.21%。优化后的方法既能通过定性分析避免假阳性的风险，又保持了较高的回收率和精密度、较宽的线性范围、较低的检出限，可以更好、更准确地用于小麦粉中脱氧雪腐镰刀菌烯醇的检测。

Abstract:

The determination of deoxynivalenol in wheat flour by high performance liquid chromatography was studied in this paper. The detection of deoxynivalenol in wheat flour was very important for the storage and quality control of wheat flour, and the chromatographic conditions of immunoaffinity-high performance liquid chromatography, specified by national standard-GB 5009-2016, had a risk of false positives for the determination of deoxynivalenol in wheat flour. Conditions of liquid chromatography ,optimized based on national standard, were as follows: mobile phase, methanol-water (25:75, V/V); detection wavelength, 240 nm; scanning range,190 nm~400nm.In addition, the qualitative analysis was also studied using full wavelength scanning by diode array detector. The results showed that the recovery was 91.7% ~ 97.2% and the detection limit was 20 μg/kg, the linear correlation coefficient was 0.99989 in the concentration range of 50 ng/mL ~ 5000 ng/mLwith the precision of 0.83% ~ 3.21%. Consequently, the optimized method could not only avoid the risk of false positives by qualitative analysis, but maintain a high recovery rate and precision, wide linear range and low detection limit, which could be better and more accurately used for the detection of deoxynivalenol in wheat flour.