地龙药材中黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)的含量检测

Detection of aflatoxins B_(1),B_(2),G_(1),G_(2)in Pheretima materials

目的建立超声萃取-免疫亲和柱净化-柱后光化学衍生法高效液相色谱法同时测定地龙药材中黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)含量的测定方法。方法样品粉碎过120目筛后,采用70%甲醇超声处理30min,经免疫亲和柱净化、高效液相色谱分离、光化学柱后衍生,通过荧光检测器测定4种黄曲霉毒素的含量。结果线性范分别为:黄曲霉毒素B_(1)0.0104~0.0520ng(r=0.9999)、黄曲霉毒素B_(2)0.0038~0.0190ng(r=0.9998)、黄曲霉毒素G_(1)0.0108~0.0540ng(r=0.9998)、黄曲霉毒素G_(2)0.0038~0.0190ng(r=0.9998),线性关系良好。检出限分别为0.43、0.15、0.43、0.15μg·kg^(-1)。回收率在90.42~99.47%之间,RSD≤3.2%(n=6)。结论该方法操作简便,灵敏度高、重复性好、结果准确,可用于地龙中黄曲霉毒素含量的测定。

OBJECTIVE To establish HPLC method for the simultaneous determination of aflatoxin B_(1),B_(2),G_(1)and G_(2)in Pheretima by ultrasonic extraction-immunoaffinity column clean-up and post-column photochemical derivatization.METHOD The sample was crushed and passed through a 120-mesh sieve and extracted with 70%methanol for tiirty minutes and purification by immunoaffinity columns,aflatoxin B_(1),B_(2),G_(1)and G_(2)in samples were anlyzed by HPLC-FLD with post-column photochemical derivatization.RESULTS The linearity of aflatoxin B_(1)was at 0.0104~0.0520ng(r=0.9999),aflatoxin B_(2)was at 0.0038~0.0190ng(r=0.9998),Aflatoxin G_(1)was at 0.0108~0.0540ng(r=0.9998),Aflatoxin G_(2)was at 0.0038~0.0190ng(r=0.9998),the detection limits were 0.43、0.15、0.43、0.15μg·kg^(-1),respectively.The recoveries ranged from 90.42%to 99.47%with the RSD≤3.2%(n=6).CONCLUSION The method is simple,sensitive,reproducible and accurate,This method is suitable for the determination of AF in Pheretima.