A method for isolating 1-hydroxypyrene from urine is described. The presence of 1-hydroxypyrene in urine was identified by fluorescence excitation and emission scanning after HPLC-separation. 1-Hydroxypyrene could be detected in the urine of rats following oral administration of as little as 0.5 microgram pyrene. The dose-dependence of 1-hydroxypyrene in urine was evident after a wide range of pyrene dosing. After therapeutical coal tar treatment of dermatological patients the enhanced excretion of 1-hydroxypyrene was highly significant. Employees of a creosote impregnating plant showed an excretion pattern of 1-hydroxypyrene which could be related to their work. 1-Hydroxypyrene in urine of non-exposed people was very low, but detectable. It is suggested that the method reported is suitable for the assessment of uptake of man to pyrene, a compound that is commonly present in work environments which are associated with pollution of polycyclic aromatic hydrocarbons.
An HPLC method for the determination of S-phenyl-N-acetylcysteine in urine is described. The sensitivity is 6 mumol/L (CV = 9%) urine. Exposure of rats to six different concentrations of benzene, ranging from 0-30 ppm, was highly associated with urinary excretion of S-phenyl-N-acetylcysteine (r = 0.86) and with total phenol (r = 0.81). A background level of phenol was found in urine of both non-exposed rats and of non-exposed referents. However, no background excretion of S-phenyl-N-acetylcysteine was found, either in rats or in humans. In urine of exposed rats, the level of S-phenyl-N-acetylcysteine was approximately five times lower than the phenol level. Workers occupationally exposed to benzene, showing high levels of urinary phenol, revealed low concentrations of urinary S-phenyl-N-acetylcysteine. The biological monitoring of industrial exposure to benzene by determination of S-phenyl-N-acetylcysteine in urine is not better than the determination of phenol in urine.
3-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 mumol 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 the Salmonella typhimurium strain TA98 in the presence of S9 mix and beta-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 the non-specific mutagenicity assay.
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