检测葡萄酒中赭曲霉毒素A的SPE-HPLC方法优化

The optimized procedure of solid-phase extraction with HPLC for the determination of ochratoxin A in wine

从固相萃取的上样体积、淋洗液体积以及高效液相色谱的洗脱程序等方面,优化并建立了检测葡萄酒中赭曲霉毒素A的（SPE-HPLC）方法。采用10mL酒样与10mL水等体积混合上样,使用2mL水淋洗后再用2mL甲醇/水（60：40,v/v）溶液的最佳淋洗条件,HPLC检测首次采用梯度洗脱,优化后方法的加标回收率为94.6%-99.5%,相对标准偏差为0.36%-3.01%。6个市售葡萄酒样品检测表明,OTA阳性率为66.7%,平均含量为0.46μg/L,证明本方法能够排除杂质峰干扰,提高准确度,可以满足葡萄酒中赭曲霉毒素（OTA）的定量检测要求。

An assay procedure of solid-phase extraction（SPE）with high performance liquid chromatography（HPLC）for the detection of ochratoxin A（OTA） in wine was optimized and established.The optimized parameters included sample volume passed though the C18 cartridge and washing solution in SPE treatment,and HPLC elution.For sample volume,10mL wine diluted with 10mL water were pre-concentrated on the C18 column.Then 2mL water was used for sample washing,followed by 2mL methanol-water solution（60/40,v/v） for further washing.For the first time,a gradient elution was used in detecting OTA in HPLC analysis.The optimized SPE-HPLC procedure improved that the recovery were between 94.6% and 99.5%,with relative standard deviations between 0.36% and 3.01%.This method was applied for the determination of OTA from 6 commercial wine samples.The percentage of OTA existing in all wine samples was 66.7%,and its content averaged to be 0.46μg/L.The SPE-HPLC procedure presented in this study should meet the needs of the quantitative measurement of OTA in wine due to the eliminated interference of impurity peak and improved accuracy of the method.