TY - JOUR
T1 - Dual-label high-performance liquid chromatographic assay for femtomole levels of benzo[a]pyrene metabolites
AU - Murphy, Sharon E.
AU - Hecht, Stephen S.
PY - 1985/5/1
Y1 - 1985/5/1
N2 - A dual-label HPLC assay to measure femtomole quantities of ethyl acetate-extractable [3H]benzo[a]pyrene metabolites was developed. 14C-Labeled metabolites of benzo[a]pyrene formed by rat liver 9000g supernatant were used as both internal standards and chromatographic markers. The percentage deviation between assays was determined to be between 11 and 13% for 9,10-dihydro-9,10-dihydroxybenzo[a]pyrene, 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene, benzo[a]pyrene-3,6-quinone, benzo[a]pyrene-1,6-quinone, and 9-hydroxybenzo[a]pyrene, 22% for 4,5-dihydro-4,5-dihydroxybenzo[a]pyrene, and less than 5% for 3-hydroxybenzo[a]pyrene. The detection limit of this assay was between 3 and 10 fmol per metabolite. The application of this technique to the metabolism of [3H]benzo[a]pyrene by microsomes of hamster and human oral cavity tissue is described.
AB - A dual-label HPLC assay to measure femtomole quantities of ethyl acetate-extractable [3H]benzo[a]pyrene metabolites was developed. 14C-Labeled metabolites of benzo[a]pyrene formed by rat liver 9000g supernatant were used as both internal standards and chromatographic markers. The percentage deviation between assays was determined to be between 11 and 13% for 9,10-dihydro-9,10-dihydroxybenzo[a]pyrene, 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene, benzo[a]pyrene-3,6-quinone, benzo[a]pyrene-1,6-quinone, and 9-hydroxybenzo[a]pyrene, 22% for 4,5-dihydro-4,5-dihydroxybenzo[a]pyrene, and less than 5% for 3-hydroxybenzo[a]pyrene. The detection limit of this assay was between 3 and 10 fmol per metabolite. The application of this technique to the metabolism of [3H]benzo[a]pyrene by microsomes of hamster and human oral cavity tissue is described.
KW - HPLC, carcinogen metabolites
KW - benzo[a]pyrene
KW - carcinogen metabolism
KW - chemical carcinogenesis
KW - dual-label analysis
UR - http://www.scopus.com/inward/record.url?scp=0021864404&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021864404&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(85)90567-6
DO - 10.1016/0003-2697(85)90567-6
M3 - Article
C2 - 4025808
AN - SCOPUS:0021864404
SN - 0003-2697
VL - 146
SP - 442
EP - 447
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -