Combined Analysis of r-1,t-2,3,c-4-Tetrahydroxy-1,2,3,4-Tetrahydrophenanthrene and 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanol in Smokers' Plasma

Abstract

Polycyclic aromatic hydrocarbons (PAH) and tobaccospecific nitrosamines, such as 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), are widely accepted to be two important types of lung carcinogens in cigarette smoke. In this study, we have developed a method to estimate individual uptake of these compounds by quantifying r -1,t -2,3,c -4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene (PheT) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in 1 mL of smokers' plasma. PheT and NNAL are biomarkers of PAH and NNK uptake, respectively. [D 10 ]PheT and [pyridine-D 4 ]NNAL were added to plasma as internal standards. The plasma was treated with B-glucuronidase to release any conjugated PheT and NNAL. The analytes were enriched by solid-phase extraction on a mixed mode cation exchange cartridge and the PheT fraction was further purified by high-performance liquid chromatography. The appropriate fractions were analyzed by gas chromatography-negative ion chemical ionization-mass spectrometry for PheT and liquid chromatography-electrospray ionization-mass spectrometry for NNAL. The method was sensitive (limits of quantitation: PheT, 13 fmol/mL; NNAL, 3 fmol/mL), accurate, and precise. Levels of PheT and NNAL in plasma from 16 smokers averaged 95 F 71 and 36 F 21 fmol/mL, respectively, which are f1% to 2% of the amounts found in urine. This method should be useful in molecular epidemiology studies of carcinogen uptake and lung cancer in smokers. (Cancer Epidemiol Biomarkers Prev 2006;15(8):1490 -4)

Metabolism of [D10]Phenanthrene to Tetraols in Smokers for Potential Lung Cancer Susceptibility Assessment: Comparison of Oral and Inhalation Routes of Administration

“…Quantitation of plasma and urinary [D 10 ]PheT were accomplished by GCnegative ion chemical ionization-MS/MS with selected reaction monitoring using [ 13 C 6 ]PheT as the internal standard as reported previously (Carmella et al, 2006;Zhong et al, 2011b). Table 1.…”

“…It had Ͼ99% chemical purity and 98% isotopic purity (Zhong et al, 2011b (Hecht et al, 2008). [D 10 ]PheT was prepared by in vitro metabolism of [D 10 ]Phe, obtained from Cambridge Isotope Laboratories, Inc., as described previously (Carmella et al, 2006). 1-Hydroxyphenanthrene (1-HOPhe; synthesized), 2-HOPhe (synthesized), 3-HOPhe (Chiron AS, Trondheim, Norway), 4-HOPhe (Chiron), and 9-HOPhe (Sigma-Aldrich, St. Louis, MO) were either synthesized by Drs.…”

“…[D 10 ]PheT in urine and plasma were analyzed by gas chromatography (GC)-negative ion chemical ionization-tandem mass spectrometry (MS/MS) as described previously, without the resolution of enantiomers (Carmella et al, 2006;Zhong et al, 2011b).…”

“…Our group has developed methods for quantitation of total r-1,t-2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene [PheT; the sum of Phe-(1R,2S,3R,4S)-tetraol (3) and Phe-(1S,2R,3S,4R)tetraol (6) in Fig. 1] in human urine and plasma Carmella et al, 2006). We have demonstrated correlations of urinary PheT with its bay region diol epoxide-derived enantiomer 3 and with the corresponding tetraol derived from BaP (Hochalter et al, 2011).…”

Syntheses of isotope‐labeled tobacco‐specific nitrosamines and their metabolites

“…For example, [pyridine-d4] NNAL has been used as an internal standard, but detailed description of its synthesis has not been given. 20 In addition, the unlabeled 5-methyl-N'-nitrososonornicotine was used as an internal standard for the quantification of total urinary NNN. 21 However, isotope-labeled internal standards are required for accurate quantification of TSNA and metabolites in biological fluids of subjects who use tobacco products.…”