Quantitative determination of hydroxy polycylic aromatic hydrocarbons as a biomarker of exposure to carcinogenic polycyclic aromatic hydrocarbons.
A high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) method was developed for quantitative analysis of hydroxy polycyclic aromatic hydrocarbons (OH-PAHs). Four hydroxy metabolites of known and suspected carcinogenic PAHs (benzo[a]pyrene (B[a]P), benz[a]anthracene (B[a]A), and chrysene (CRY)) were selected as suitable biomarkers of PAH exposure and associated risks to human health. The analytical method included enzymatic deconjugation, liquid - liquid extraction, followed by derivatization with methyl-N-(trimethylsilyl) trifluoroacetamide and instrumental analysis. Photo-induced oxidation of target analytes - which has plagued previously published methods - was controlled by a combination of minimizing exposure to light, employing an antioxidant (2-mercaptoethanol) and utilizing a nitrogen atmosphere. Stability investigations also indicated that conjugated forms of the analytes are more stable than the non-conjugated forms. Accuracy and precision of the method were 77.4-101% (<4.9% RSD) in synthetic urine and 92.3-117% (<15% RSD) in human urine, respectively. Method detection limits, determined using eight replicates of low-level spiked human urine, ranged from 13 to 24pg/mL. The method was successfully applied for analysis of a pooled human urine sample and 78 mouse urine samples collected from mice fed with PAH-contaminated diets. In mouse urine, greater than 94% of each analyte was present in its conjugated form.
AXYS Analytical Services Ltd, 2045 Mills Road West, Sidney, BC, V8L 5X2, Canada. Electronic address: mwoudneh@axys.com.
Million B
MB
Woudneh
AXYS Analytical Services Ltd, 2045 Mills Road West, Sidney, BC, V8L 5X2, Canada.
Jonathan P
JP
Benskin
AXYS Analytical Services Ltd, 2045 Mills Road West, Sidney, BC, V8L 5X2, Canada.
Richard
R
Grace
AXYS Analytical Services Ltd, 2045 Mills Road West, Sidney, BC, V8L 5X2, Canada.
M Coreen
MC
Hamilton
Arcadis U.S., Inc., One Executive Drive, Suite 303, Chelmsford, MA 01824, United States.
Brian H
BH
Magee
Arcadis U.S., Inc., 1 South Church Avenue, Suite 1120, Tucson, Az 85701, United States.
Glenn C
GC
Hoeger
Arcadis U.S., Inc., One Executive Drive, Suite 303, Chelmsford, MA 01824, United States.
Norman D
ND
Forsberg
AXYS Analytical Services Ltd, 2045 Mills Road West, Sidney, BC, V8L 5X2, Canada.
John R
JR
Cosgrove
D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D002273:Carcinogens / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D051379:Mice; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis*
D015415:Biomarkers; D002273:Carcinogens; D011084:Polycyclic Aromatic Hydrocarbons
Antioxidants; Biomonitoring; GC–HRMS; OH-PAHs; Stability; Urine