免疫亲和色谱柱-高效液相色谱法测定饲料中赭曲霉毒素A的研究

Study on the determination of ochratoxin A in feed by immunoaffinity chromatography column and high performance liquid chromatography

本文建立了饲料中赭曲霉毒素A(OTA)的免疫亲和柱净化-高效液相色谱荧光检测方法。样品用甲醇-水(体积比80∶20)提取,通过免疫亲和柱富集和净化,以乙腈和2%乙酸溶液为流动相(体积比44∶56),Agilent C18色谱柱分离,荧光检测器检测(λex-332 nm,λem=460 nm)。OTA的线性范围为0.1～100.0 ng/mL,r=0.99989。方法检测限为0.2μg/kg,定量限为0.6μg/kg。样品中赭曲霉毒素A的加标回收率为75%～92%,相对标准偏差为1.65%～3.61%。

A method was established for the determination of ochratoxin A（OTA） in feed by high performance liquid chromatography （HPLC） technique, with immunoaffinity column （IAC）pre-separation and fluorescence detection （FLD）. Feed samples were first extracted with methanol-water （80：20, V/V） solution.Then,the extract was pre-separated and concentrated by the using of IAC.The optimized mobile phase was mixture of acetonitrile and 2 % acetic acid （44：56, V/ V） ,the analysis was performed on an Agilent C18 column, and quantified with fluorescence detection （λex=332 nm, λem= 460 nm）.Linear response for OTA was observed in the range of 0.1 - 100.0 ng/mL （r=0.99989）.The limit of detection （LOD） and the quantization （LOQ） were 0.2 ug/kg and 0.6 ug/kg,respectively.Recoveries of 75% - 92% were observed for the feed spiked with OTA,and the relative standard deviation was 1.65 % - 3.61%.