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目的建立一种全自动在线免疫亲和柱固相萃取-高效液相色谱(online immunoaffinity solid phase extraction-high performance liquid chromatography,online-IA-SPE-HPLC)快速测定食品中黄曲霉毒素(aflatoxins,AFT)。方法样品经乙腈-水提取后,经可重复使用的在线免疫亲和小柱进行富集与净化,经C_(18)色谱柱(150mm×4.6 mm,5μm)进行分离,用特定流动相进行洗脱,用电化学衍生KOBRA~?Cell荧光检测器进行检测,激发波长为362 nm,发射波长为455 nm,同时进行线性范围、精密度、准确度和加标回收率等方法学验证。结果对于大米、食醋、酱油和花生油等基质,该方法对AFT B_1的检出限均为0.035μg/kg,低于我国现行食品安全标准对AFT B_1的限量要求;AFT B_1及AFT G1在25~500 ng/L范围内具有良好的线性,AFT B_2及AFT G_2在6.25~125 ng/L范围内具有良好的线性,相关系数均为0.9999;在低、中和高3水平加标下大米、食醋、酱油和花生油中的4种黄曲霉毒素回收率在91.16%~109.99%之间,精密度为0.52%~4.39%。结论该方法的前处理快速、简易,方法重现性好、灵敏度高,适用于食品中AFT的高通量检测。
OBJECTIVE: To establish a rapid and sensitive method for the determination of aflatoxins (AFT) in food by online immunoaffinity solid phase extraction-high performance liquid chromatography (online-IA-SPE-HPLC) ). Methods The sample was extracted with acetonitrile - water, and then was reused on an online immunoaffinity cartridge for enrichment and purification. The sample was separated on a C 18 column (150 mm × 4.6 mm, 5 μm) and washed with a specific mobile phase Off and electrochemically derivatized with a KOBRA ~? Cell fluorescence detector. The excitation wavelength was 362 nm and the emission wavelength was 455 nm. Simultaneously, methodological validation such as linear range, precision, accuracy and spike recovery was performed. Results The detection limits of AFT B_1 for rice, vinegar, soy sauce and peanut oil were all 0.035 μg / kg, which were lower than the limit of AFT B_1 in our country. AFT B_1 and AFT G1 were 25 ~ 500 ng / L, AFT B 2 and AFT G 2 have good linearity in the range of 6.25-125 ng / L with the correlation coefficients of 0.9999. Under the three levels of low, medium and high, The recoveries of four aflatoxins in vinegar, soy sauce and peanut oil ranged from 91.16% to 109.99% with the precision of 0.52% -4.39%. Conclusion The pretreatment of this method is rapid, simple, reproducible and sensitive. It is suitable for the high-throughput detection of AFT in food.