Abstract3‐hydroxy‐benzo(a)pyrene (3‐OH‐B(a)P) and mutagenic activity in rat urine were determined after the oral administration of benzo(a)pyrene given in three repeated doses of 10, 20 and 50 μmol kg−1. The procedure for the determination of 3‐OH‐B(a)P consisted of enzymic hydrolysis, separation and HPLC‐analysis. The mutagenic activity of concentrated urine samples was assayed with theSalmonella typhimuriumstrain TA98 in the presence of S9 mix and β‐glucuronidase. The urinary excretion of 3‐OH‐B(a)P and mutagens showed a correlation and both increased dose‐dependently during the sampling period of 6 days. Data indicated that 3‐OH‐B(a)P can be regarded as a reliable representative of all urinary (pre)‐mutagens derived from benzo(a)pyrene and exposure of rats to benzo(a)pyrene could be detected with greater sensitivity by the HPLC assay of 3‐OH‐B(a)P than with
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