建立了一种新的测定酿酒葡萄中赭曲霉毒素A含量的方法——"葡萄皮-SPE-HPLC"法:将酿酒葡萄剥皮后用0.1 mol/L磷酸-二氯甲烷(1∶10)提取;取10 mL提取液进样固相萃取小柱,0.1 mol/L磷酸、双蒸水淋洗,乙酸乙酯洗脱后吹干,流动相溶解;高效液相色谱法测定条件为C18反相柱分离,乙腈-水-乙酸(体积比99∶99∶2)为流动相,荧光检测器(激发波长330 nm,发射波长460 nm)检测。该方法回收率为102%~109%,变异系数为0.1%~2.0%;用此方法检测贺兰山东麓"玉泉营"和"万义山庄"两个产地酿酒葡萄中OTA含量,结果介于5~10μg/kg,均有一定程度赭曲霉毒素A污染。

The new method for determination of Ochratoxin A in grape established in this paper named"PericarpSPE-HPLC". The fresh wine grape was peeled. Ochratoxin A was extracted from grape pericarp with 0. 1M phosphoric acid- dichloromethane( 1∶ 10). After C18solid phase extraction column( C18-SPE) has activated with 5mL Methanol,5mL double diluted water,and 5mL Sodium hydrogen carbonate( 30g / L),the above extract 10mL was firstly brought into C18-SPE column. Then the C18-SPE column was washed with 2mL phosphoric acid( 0. 1mol / L) and2mL double distilled water respectively and was eluted with 5mL ethyl acetate subsequently. The eluent was dried with N2gas. The residue was dissolved with 0. 2 mL of mobile phase for determination. The chromatographic column was C18 reversed-phase column( 150 mm × 4. 6 mm i. d.) for separation. Mobile phase was acetonitrile-water-acetic acid( 99∶ 99∶ 2). A high performance liquid chromatography was determined with fluorescence detection. The excitation wavelength was 330nm. The emission wavelength was 460 nm. The recoveries of this method were between 102. 5%and 109. 7%,and coefficient of variation varied from 0. 14% to 2. 07%. Results from analysis with "Pericarp-SPEHPLC"method showed that Ochratoxin A in two grape samples "yuquanying"and "wanyi heights"were 5 ~ 10 μg /kg.