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Evaluation of Methods for Quantitation of Aflatoxin-Albumin Adducts and Their Application to Human Exposure Assessment¹

Unit of Mechanisms of Carcinogenesis, International Agency for Research on Cancer, 150 cours Albert Thomas, 69372 Lyon Cedex 08, France

Aflatoxin (AF) albumin adducts are found in peripheral blood after exposure to aflatoxin B₁ (AFB₁) and the measurement of these adducts is potentially a useful tool in the epidemiological study of the role of AFB₁ in the etiology of liver cancer. Three complementary approaches to the quantitation of AF-albumin adducts are described: (a) enzymelinked immunosorbent assay (ELISA) performed directly on intact albumin (direct ELISA); (b) ELISA performed on an albumin hydrolysate (hydrolysis ELISA); (c) high-performance liquid chromatographic fluorescence detection of AF-lysine adduct after albumin hydrolysis and immunoaffinity purification. These techniques have been validated by direct comparison with rat albumin samples modified to a known extent. Detection limits of 100, 5.0, and 5.0 pg AF/mg human albumin were determined for the three methods, respectively. Samples obtained from individuals from Thailand, The Gambia, Kenya, and France have been used to validate the measurement of AF-albumin adducts by these three methods. Levels of 7 to 338 pg AF/mg albumin were observed in the former two countries while no adducts were detected in samples from France. The relative properties of the three assays, with special regard to their application in epidemiological studies, are considered. A combination of the hydrolysis ELISA for large scale screening followed by confirmatory analyses in positive samples by high-performance liquid chromatographic fluorescence is suggested as an optimum methodology.

¹ The work reported here was partially supported by the International Programme on Chemical Safety of the World Health Organization.

² To whom requests for reprints should be addressed.

³ Supported by a fellowship from l'Association de Recherche Contre le Cancer (France), which enabled this work to be performed. Permanent address: Department of Chemical Etiology, Cancer Institute, Chinese Academy of Medical Sciences, Beijing, People's Republic of China.

⁴ Supported by Swiss National Foundation Grant 83.518.187. Present address: Institut für Pharmakologie und Toxikologie, Universität Würzburg, Verbacherstrasse 9, D.8700 Würzburg, Federal Republic of Germany.

Received 7/ 6/89. Revised 9/27/89. Accepted 10/11/89.

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