Groopman J D, Hasler J A, Trudel L J, Pikul A, Donahue P R, Wogan G N
Department of Environmental Health Sciences, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland 21205.
Cancer Res. 1992 Jan 15;52(2):267-74.
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.
分子剂量测定方法的发展将简化对癌症高危人群的识别。已设计出一种结合单克隆抗体免疫亲和色谱法/高效液相色谱法，用于分离和定量大鼠尿液样本中的黄曲霉毒素 - DNA加合物及其他代谢物。我们报告了针对黄曲霉毒素B1、黄曲霉毒素Q1、黄曲霉毒素G1、黄曲霉毒素醇和黄曲霉毒素M1产生的11种不同单克隆抗体，以及这些抗体在多克隆抗体亲和色谱技术中的应用。使用多克隆抗体亲和柱处理给药动物的大鼠尿液，总黄曲霉毒素代谢物的90%至95%可与柱子结合并分离出来。对这些分离出的黄曲霉毒素进行分析免疫亲和色谱/高效液相色谱分析表明，大鼠尿液中超过55%的黄曲霉毒素为黄曲霉毒素 - 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，分别占总黄曲霉毒素的1.5%、9.6%、1.8%、34.5%、8.0%和1.0%。此外，通过高氯酸消化黄曲霉毒素 - N7 - 鸟嘌呤峰，将其转化为鸟嘌呤来确认其身份。检测了大鼠尿液中黄曲霉毒素 - N7 - 鸟嘌呤的排泄量，以评估其作为肝脏中DNA加合物形成标志物的效用，发现尿液排泄呈剂量依赖性，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 - 鸟嘌呤排泄量的比较显示相关系数为0.98。除了核酸加合物排泄数据外，还对尿液中的黄曲霉毒素M1和黄曲霉毒素P1作为分子剂量计进行了评估。发现黄曲霉毒素M1是一个优秀的标志物，而尿液中黄曲霉毒素P1排泄量与剂量之间未发现线性关系。
