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目的建立高效液相色谱-荧光检测法,测定黄曲霉毒素B1(AFB1)染毒大鼠肝脏组织与血清样品中AFB1的含量。方法无特定病原体(SPF)的Fischer 344雄性大鼠10只,随机分为AFB1染毒组和溶剂对照组,采用腹腔注射AFB1的方式染毒4周,每周染毒3次,剂量为0.2 mg/kg,对照组注射同等剂量的玉米油。染毒结束后,采用高效液相色谱法(HPLC)检测大鼠血清和肝脏中AFB1的含量。HPLC采用C18色谱柱分离,水(含有0.2%甲酸)、甲醇和乙腈为流动相,梯度洗脱,荧光检测器在激发波长设为365 nm、发射波长设为435 nm的条件下检测AFB1的量。结果AFB1的洗脱出峰时间约为8.748 min,检出限为0.9μg/L,回收率为80.6%~92.5%。染毒组大鼠血清中AFB1平均含量为17.49μg/L,肝脏中AFB1平均含量为26.80μg/kg,溶剂对照组大鼠血清和肝脏中未检测出AFB1。结论该方法能准确、灵敏的检测出AFB1染毒大鼠血清和肝脏组织中的暴露剂量。
OBJECTIVE To establish a method for the determination of AFB1 in liver tissue and serum of aflatoxin B1 (AFB1) -treated rats by high performance liquid chromatography-fluorescence detection. Methods Ten Fischer 344 male rats without specific pathogen (SPF) were randomly divided into two groups: AFB1 group and solvent control group. The rats were injected AFB1 intraperitoneally for 4 weeks and exposed to 0.2 mg / / kg, the control group injected the same dose of corn oil. After the exposure, the contents of AFB1 in serum and liver of rats were detected by high performance liquid chromatography (HPLC). HPLC was separated on a C18 column with water (containing 0.2% formic acid) and methanol and acetonitrile as mobile phases. The fluorescence detector was used to detect the amount of AFB1 at an excitation wavelength of 365 nm and an emission wavelength of 435 nm . Results The peak elution time of AFB1 was 8.748 min, the detection limit was 0.9 μg / L and the recovery was 80.6% -92.5%. The average content of AFB1 in serum was 17.49μg / L and the average content of AFB1 in liver was 26.80μg / kg in the treated group. AFB1 was not detected in serum and liver of the solvent control group. Conclusion The method can accurately and sensitively detect the exposure dose in the serum and liver tissues of AFB1-exposed rats.