TY - JOUR
T1 - Detection of metabolites of polycyclic aromatic hydrocarbons in human urine
AU - Weston, Ainsley
AU - Bowman, Elise D.
AU - Carr, Paula
AU - Rothman, Nathaniel
AU - T.strickland, Paul
N1 - Funding Information:
'Laboratory of Human Carcinogenesis and4Environmental Epidemiology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 and3Scbool of Hygiene and Public Health, Johns Hopkins University, Baltimore, MD 21205, USA 2To whom correspondence should be addressed at present address: Department of Community Medicine, Mount Sinai School of Medicine, 1 Gustave L.Levy Place, Box 1057, New York, NY 10029, USA 3Paula Carr is a summer student through the NCI-MARC Program at NIH from Toogaloo College, Toogaloo, MI, USA
PY - 1993/5
Y1 - 1993/5
N2 - A non-invasive assay has been developed for the recovery of r-7,t-8,t-9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[α]-pyrene (BP-7,10/8,9-tetrol) from human urine. This tetrol is excreted as a metabolite of benzo[α]pyrene (BP) in a process catalyzed by cytochrome P450 enzymes and epoxide hydrolases. Urine was hydrolysed to release activated benzo[α]-pyrene-diol-epoxides covalently bound to macromolecular species or conjugated tetrols. The relatively non-polar organic molecules from urine hydrolysates were collected on octadecasilane chromatography columns (Sep-Paks). Materials eluted in solvent (80% CH3OH), were further purified on immunoaffinity columns with antibodies raised against anti-N2-[10(7,8,9-trihydroxy-7,8,9,10-tetra-hydrobenzo[α]pyrenyl)]-guanosine. HPLC was then used to isolate BP-7,10/8,9-tetrol, which was quantitated by synchronous fluorescence spectroscopy (SFS). This assay detected 0.24-3.12 pmol BP-7, 10/8,9-tetrol per ml urine (limit of detection 0.01 pmol/ml, given 10 ml urine), in four study subjects. Reproducibility was assessed by adding tritium labeled BP-7, 10/8,9-tetrol (1500 fmol) to a urine sample previously identified to contain the tetrol at levels below the limit of detection of the fluorescence assay; a recovery of >30% of the added radioactivity was achieved (510 ± 64 fmol, mean ± SD, n = 3). Because HPLC alone was not sufficient to isolate materials for quantitation by SFS directly from human urine, immunoaffinity chromatography was found to be a necessary preparatory step in BP-7,10/8,9-tetrol isolation. These data demonstrate the presence of tetrahydrotetrol metabolites of BP in human urine and suggest that measurement of BP-7,10/8,9-tetrol and other polycyclic aromatic hydrocarbon-tetrols may prove to be valuable dosimeters of human internal exposure to polycyclic aromatic hydrocarbons.
AB - A non-invasive assay has been developed for the recovery of r-7,t-8,t-9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[α]-pyrene (BP-7,10/8,9-tetrol) from human urine. This tetrol is excreted as a metabolite of benzo[α]pyrene (BP) in a process catalyzed by cytochrome P450 enzymes and epoxide hydrolases. Urine was hydrolysed to release activated benzo[α]-pyrene-diol-epoxides covalently bound to macromolecular species or conjugated tetrols. The relatively non-polar organic molecules from urine hydrolysates were collected on octadecasilane chromatography columns (Sep-Paks). Materials eluted in solvent (80% CH3OH), were further purified on immunoaffinity columns with antibodies raised against anti-N2-[10(7,8,9-trihydroxy-7,8,9,10-tetra-hydrobenzo[α]pyrenyl)]-guanosine. HPLC was then used to isolate BP-7,10/8,9-tetrol, which was quantitated by synchronous fluorescence spectroscopy (SFS). This assay detected 0.24-3.12 pmol BP-7, 10/8,9-tetrol per ml urine (limit of detection 0.01 pmol/ml, given 10 ml urine), in four study subjects. Reproducibility was assessed by adding tritium labeled BP-7, 10/8,9-tetrol (1500 fmol) to a urine sample previously identified to contain the tetrol at levels below the limit of detection of the fluorescence assay; a recovery of >30% of the added radioactivity was achieved (510 ± 64 fmol, mean ± SD, n = 3). Because HPLC alone was not sufficient to isolate materials for quantitation by SFS directly from human urine, immunoaffinity chromatography was found to be a necessary preparatory step in BP-7,10/8,9-tetrol isolation. These data demonstrate the presence of tetrahydrotetrol metabolites of BP in human urine and suggest that measurement of BP-7,10/8,9-tetrol and other polycyclic aromatic hydrocarbon-tetrols may prove to be valuable dosimeters of human internal exposure to polycyclic aromatic hydrocarbons.
UR - http://www.scopus.com/inward/record.url?scp=0027318241&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027318241&partnerID=8YFLogxK
U2 - 10.1093/carcin/14.5.1053
DO - 10.1093/carcin/14.5.1053
M3 - Article
C2 - 8504465
AN - SCOPUS:0027318241
SN - 0143-3334
VL - 14
SP - 1053
EP - 1055
JO - Carcinogenesis
JF - Carcinogenesis
IS - 5
ER -