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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,1.7μm)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1μL,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1.15mL,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。
OBJECTIVE To establish a rapid and quantitative method for the determination of ochratoxin A (OTA) in foodstuffs by immunoaffinity column cleanup and ultra performance liquid chromatography. Methods After the sample was extracted with methanol: water (80:20, v: v), it was purified by immunoaffinity column and concentrated. The residue was separated on a Waters Acquity UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm) : Glacial acetic acid (100: 98: 2, v: v: v) as the mobile phase with a flow rate of 0.2 mL / min and an injection volume of 1 μL. The detection wavelength was 310 nm with an excitation wavelength of 465 nm. Aspergillus toxin A retained a volume of less than 1.15 mL, from sample pretreatment to analysis of less than 45 min throughout the process. Results The detection limit of ochratoxin A was 0.25 pg based on the 3-fold peak response of signal-to-noise ratio and linearly correlated in the range of 1 ~ 50 pg. The correlation coefficient R2 was 0.998. The recoveries were 81.5% -101.2%, 81.3% -85.5%, 87.8% -97.5% and the precision was 3.3% -6.4% in wheat, corn and paddy rice samples, respectively. Conclusion The method is simple and rapid, high sensitivity, good reproducibility and less solvent. It is suitable for the rapid determination of ochratoxin A in food.