粮食中赭曲霉毒素A的快速定量检测-超高效 液相色谱法

Rapid Analysis of Ochratoxin A in Grain by Ultra Performance Liquid Chromatography

中文关键词:  粮食  赭曲霉毒素A  超高效液相色谱法  免疫亲和柱

英文关键词:grain  ochratoxin A  ultra performance liquid chromatography  immuno-affinity column

中文摘要:

      目的 建立了免疫亲和柱净化-超高效液相色谱法快速定量测定粮食中赭曲霉毒素A(ochratoxin A, OTA)的检测方法。方法 样品经甲醇:水(80:20, v:v)提取后, 用免疫亲和柱净化、浓缩, Waters Acquity UPLC BEH C18色谱柱(50 mm×2.1 mm, 1.7 μm)分离, 以乙腈:水:冰醋酸(100:98:2, v:v:v)为流动相, 流速为0.2 mL/min, 进样量为1 μL, 荧光检测器检测, 激发波长为310 nm发射波长为465 nm, 赭曲霉毒素A的保留体积小于1.15 mL, 从样品前处理到分析整个过程小于45 min。结果 根据3倍信噪比的峰响应值, 确定赭曲霉毒素A的检出限为0.25 pg, 在1~50 pg范围内呈线性相关, 相关系数R2为0.998。在小麦、玉米、稻谷样品中加标回收率分别为81.5%~101.2%, 81.3%~85.5%, 和 87.8%~97.5%, 精密度为3.3%~6.4%。结论 本方法简便快速, 灵敏度高, 重现性好, 溶剂用量少, 适用于粮食中赭曲霉毒素A的快速测定。

英文摘要:

      Objective To establish a rapid and environment-friendly analytic method without any derivation for determination of ochratoxin A (OTA) in grain samples. Methods The samples were extracted by me-thanol/water (80:20, v:v), cleared up with the immuno-affinity column (IAC). The separation of targeted compound was performed on a Waters Acquity UPLC BEH C18 (2.1 mm×50 mm, 1.7 μm) using acetonitrile : water : acetic acid (100: 98:2, v:v:v) as mobile phase with a flow rate of 0.2 mL/min at 25 ℃. The injection volume was 1 μL and detection conditions were set at 310 nm (Em) and 465 nm (Ex) using fluorescence detector (FLD). The retention volume was less than 1.15mL, and the whole analytical time was less than 45 min. Results The detection limit of OTA was 0.25 pg. The linear detection ranges of OTA was 1~50 pg with correlation coefficients (R2) of 0.998. Recovery rates in grains (wheat, corn, and rice) spiked with OTA were in the range of 81.5%~101.2%, 81.3%~85.5%, and 87.8%~97.5%, respectively, with the relative standard deviation of 3.3%~6.4%. Conclusion This method was suitable for determination of OTA in the raw grain, with the advantages of simplicity, rapidness, sensitivity, good reproducibility and environment-friendly.