Molecular dosimetry in rat urine of aflatoxin-N7-guanine and other aflatoxin metabolites by multiple monoclonal antibody affinity chromatography and immunoaffinity/high performance liquid chromatography.
Molecular dosimetry in rat urine of aflatoxin-N7-guanine and other aflatoxin metabolites by multiple monoclonal antibody affinity chromatography and immunoaffinity/high performance liquid chromatography.
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发表时间：
1992-01
期刊：
影响因子：
11.2
通讯作者：
J. Groopman;J. Hasler;L. Trudel;A. Pikul;P. Donahue;G. Wogan
中科院分区：
文献类型：
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作者：
J. Groopman;J. Hasler;L. Trudel;A. Pikul;P. Donahue;G. Wogan
分子剂量法的发展将简化对癌症高危人群的识别。建立了一种单克隆抗体免疫亲和层析/高效液相层析相结合的方法，用于分离和定量大鼠尿液中黄曲霉毒素- dna加合物和其他代谢产物。我们报道了11种识别黄曲霉毒素B1、黄曲霉毒素Q1、黄曲霉毒素G1、黄曲霉毒素和黄曲霉毒素M1的不同单克隆抗体的产生，并将这些抗体应用于多单克隆抗体亲和层析技术。利用与给药动物尿液的多重单克隆抗体亲和柱，90% ~ 95%的黄曲霉毒素总代谢物可以结合到该柱上并分离出来。分析免疫亲和层析/高效液相色谱分析表明，大鼠尿液中黄曲霉毒素的55%以上为黄曲霉毒素-二氢二醇、黄曲霉毒素- n7 -鸟嘌呤、黄曲霉毒素Q1、黄曲霉毒素M1、黄曲霉毒素P1和黄曲霉毒素B1，分别占黄曲霉毒素总量的1.5%、9.6、1.8、34.5、8.0和1.0%。此外，用高氯酸消化黄曲霉毒素- n7 -鸟嘌呤峰，通过其转化为鸟嘌呤来确认其身份。测定了黄曲霉毒素- n7 -鸟嘌呤在大鼠尿液中的排泄量，以评估其作为肝脏中DNA加合物形成标记物的效用，发现尿中黄曲霉毒素- n7 -鸟嘌呤的排泄量呈剂量依赖性，相关系数为0.99。黄曲霉毒素与肝脏DNA结合的剂量依赖性残留水平与尿液中黄曲霉毒素- n7 -鸟嘌呤排泄量的相关系数为0.98。除核酸加合物排泄数据外，黄曲霉毒素M1和黄曲霉毒素P1作为尿中的分子剂量计进行评估。黄曲霉毒素M1被发现是一个很好的标志物，而黄曲霉毒素P1在尿中排泄量与剂量之间没有线性关系。
The development of molecular dosimetry methods will simplify the identification of people at high risk for cancer. A combined monoclonal antibody immunoaffinity chromatography/high performance liquid chromatography method has been devised to isolate and quantify aflatoxin-DNA adducts and other metabolites in rat urine samples. We report the production of 11 different monoclonal antibodies recognizing aflatoxin B1, aflatoxin Q1, aflatoxin G1, aflatoxicol, and aflatoxin M1 and the application of these antibodies to a multiple monoclonal antibody affinity chromatography technique. Using the multiple monoclonal antibody affinity column with rat urines obtained from dosed animals, between 90 and 95% of total aflatoxin metabolites can be bound to the column and isolated. Analytical immunoaffinity chromatography/high performance liquid chromatography analysis of these isolated aflatoxins reveals that more than 55% of the aflatoxins in rat urine are aflatoxin-dihydrodiol, aflatoxin-N7-guanine, aflatoxin Q1, aflatoxin M1, aflatoxin P1, and aflatoxin B1, accounting for 1.5, 9.6, 1.8, 34.5, 8.0, and 1.0% of the total aflatoxins, respectively. Further, a perchloric acid digestion of the aflatoxin-N7-guanine peak was used to confirm its identity by its conversion to guanine. The measurement of aflatoxin-N7-guanine excretion in rat urine was examined to assess its utility as a marker of DNA adduct formation in the liver, and a dose-dependent excretion in urine was found with a correlation coefficient of 0.99. A comparison of the dose-dependent residual levels of aflatoxin binding to liver DNA with the amount of aflatoxin-N7-guanine excreted in urine showed a correlation coefficient of 0.98. Besides the nucleic acid adduct excretion data, aflatoxin M1 and aflatoxin P1 were evaluated as molecular dosimeters in the urine. Aflatoxin M1 was found to be an excellent marker, whereas no linear relationship between dose and aflatoxin P1 excretion in urine was found.