Simultaneous analysis of naphthols, phenanthrols, and 1-hydroxypyrene in urine as biomarkers of polycyclic aromatic hydrocarbon exposure: intraindividual variance in the urinary metabolite excretion profiles caused by intervention with beta-naphthoflavone induction in the rat.
Two fluorimetric HPLC methods are described for the quantification of naphthols, phenanthrols and 1-hydroxypyrene (1-OHP) in urine specimens obtained from male Wistar rats exposed to naphthalene, phenanthrene and pyrene. The polycyclic aromatic hydrocarbons (PAHs) were given intraperitoneally, either alone (1.0 mmol/kg body weight) or as an equimolar mixture (0.33 mmol/kg), using the same dosages for repeated treatments on week 1 and week 2. Between these treatments, PAH-metabolizing activities encoded by aryl hydrocarbon (Ah) receptor-controlled genes were induced in the rats with beta-naphthoflavone (betaNF). Chromatographic separation of five phenanthrols (1-, 2-, 3-, 4-, and 9-isomers) was accomplished using two different RP C-18 columns. Despite selective detection (programmable wavelengths), the quantification limits in the urine ranged widely: 1-OHP (0.18 microg/l) <phenanthrols (0.34-0.45 microg/l) <2-naphthol (1.5 microg/l) <1-naphthol (4 micro g/l). The relative standard deviation of the methods was good, as also was the reproducibility. The molar fraction of the dose excreted in 24-h urine as naphthols (<or=4.0%), phenanthrols (<or=1.1%), and 1-OHP (<or=2.4%) was low. Urinary disposition increased differentially in betaNF-induced rats: naphthols, 9-phenanthrol (1- to-2-fold); 2-, 3-, and 4-phenanthrols (4- to 5-fold); 1-phenanthrol and 1-OHP (over 11-fold). The OH-metabolites were analyzed before and after enzymatic hydrolysis (beta-glucuronidase/arylsulfatase). The percentage excreted as a free phenol in urine varied for 1-OHP (2-11%), 1-naphthol (36-51%), 2-naphthol (59-65%), and the phenanthrols (29-94%). 1-Naphthyl- and 1-pyrenyl beta- d-glucuronide served as measures for the completeness of enzymatic hydrolysis. Characteristic differences observed in the urinary disposition of naphthalene, phenanthrene, and pyrene are described, as well as important factors (dose, metabolic capacity, relative urinary output) associated with biomarker validation. This intervention study clarifies intraindividual variation in PAH metabolism and provides useful information for the development of new methods applicable in the biomonitoring of PAH exposure in humans.
Elovaara
Eivor
E
Laboratory of Toxicokinetics and Metabolism, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, 00250, Helsinki, Finland. Eivor.Elovaara@occuphealth.fi
Väänänen
Virpi
V
Mikkola
Jouni
J
D000284:Administration, Oral; D000818:Animals; D015415:Biomarkers / Q000652:urine*; D001835:Body Weight / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis; D004359:Drug Therapy, Combination; D004784:Environmental Monitoring / Q000379:methods*; D004790:Enzyme Induction; D014644:Genetic Variation; D007274:Injections, Intraperitoneal; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008297:Male; D009284:Naphthols / Q000652:urine*; D010616:Phenanthrenes / Q000652:urine*; D011084:Polycyclic Aromatic Hydrocarbons / Q000493:pharmacokinetics / Q000633:toxicity*; D011721:Pyrenes / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D019324:beta-Naphthoflavone / Q000494:pharmacology*
D015415:Biomarkers; D009284:Naphthols; D010616:Phenanthrenes; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes; C031181:phenanthrene; D019324:beta-Naphthoflavone; D019363:Cytochrome P-450 CYP1A1; C033146:1-hydroxypyrene