Simultaneous, rapid and sensitive quantification of 8-hydroxy-2'-deoxyguanosine and cotinine in human urine

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1 Analytical and Bioanalytical Chemistry Electronic Supplementary Material Simultaneous, rapid and sensitive quantification of 8-hydroxy-2'-deoxyguanosine and cotinine in human urine by on-line solid-phase extraction LC-MS/MS: correlation with tobacco exposure biomarkers NNAL Chao-Yu Chen, Yu-Ting Jhou, Hui-Ling Lee, Yu-Wen Lin 1

2 Fig. S1 Representative MRM chromatograms of LC-MS/MS analysis for total NNAL in human urine. The concentration of NNAL was ng ml -1, and the concentration of IS was 2 ng ml -1 in the human urine 2

3 Table S1 Literature basal level of 8-OHdG, cotinine and total NNAL in urine samples from smoker or non-smoker subjects Analytes Method Level (ng/mg Creatinine ± SD) Subjects Reference 8-OHdG HPLC-ECD a 4.90 ± 1.09 Non-smoker (n = 58) [1] LC-MS/MS 6.30 ± 0.20 Non-smoker (n = 96) [2] LC-MS/MS 3.72 ± 0.18 Non-smoker (n = 75) [3] ELISA b 7.36 ± 2.50 Non-smoker (n = 627) [4] On-line SPE LC-MS/MS 3.01 ± 1.67 Non-smoker (n = 40) Our study ELISA b 7.75 ± 2.80 Smoker (n = 154) [4] HPLC-ECD a 3.68 ± 0.05 Smoker (n = 104) [5] LC-MS/MS 6.90 ± 0.30 Smoker (n = 80) [2] On-line SPE LC-MS/MS 4.03 ± 1.91 Smoker (n = 40) Our study Cotinine HPLC-MS/MS 2471 (1,662-4,393) Smoker (n = 130) [6] HPLC-UV 975 ± 919 Smoker (n = 101) [7] RIA c 703 ( ) Smoker (n = 3) [8] On-line SPE LC-MS/MS ± Smoker (n = 40) Our study Total NNAL HPLC-MS/MS 0.22 ( ) Smoker (n = 130) [6] HPLC-MS/MS 0.17 ± 0.14 Smoker (n = 101) [7] HPLC-MS/MS 0.04 ( ) Smoker (n = 23) [9] GC-MS/MS 0.06 Smoker (n = 343) [10] LC-MS/MS 0.07 ± 0.04 Smoker (n = 40) Our study a High performance liquid chromatography-electrochemical detection b Enzyme-linked immunosorbent assay c Radioimmunoassay 3

4 Table S2 Comparison of free NNAL and total NNAL levels in literatures and present study Reference Group Free NNAL Ratio c Total NNAL Ratio c (ng mg -1 of creatinine) (ng mg -1 of creatinine) This study Non-smoker a (n=40) 0.03 ± ± Smoker a (n=40) 0.04 ± ± 0.04 Al Ali R et al. [9] Non-smoker a (n=28) Smoker a (n=23) Hu et al. [7] Non-smoker a (n=40) - b - b Smoker a (n=101) 0.10 ± ± 0.14 Lee et al. [11] Non-smoker a (n=69) 0.48 ± ± Smoker a (n=57) 1.06 ± ± 0.99 Chung et al. [12] Non-smoker a (n=81) 0.14 ± ± Smoker a (n=22) 0.13 ± ±0.09 a : mean ± SD, b : not mention, c: the concentration of smoker / the concentration of non-smoker. 4

5 References 1. Svoboda P, Ko SH, Cho B, Yoo SH, Choi SW, Ye SK, Kasai H, Chung MH (2008) Neopterin, a marker of immune response, and 8-hydroxy-2'-deoxyguanosine, a marker of oxidative stress, correlate at high age as determined by automated simultaneous high-performance liquid chromatography analysis of human urine. Anal Biochem 383 (2): Harman SM, Liang L, Tsitouras PD, Gucciardo F, Heward CB, Reaven PD, Ping W, Ahmed A, Cutler RG (2003) Urinary excretion of three nucleic acid oxidation adducts and isoprostane F(2)alpha measured by liquid chromatography-mass spectrometry in smokers, ex-smokers, and nonsmokers. Free Radic Biol Med 35 (10): Huang CY, Su CT, Chung CJ, Pu YS, Chu JS, Yang HY, Wu CC, Hsueh YM (2012) Urinary total arsenic and 8-hydroxydeoxyguanosine are associated with renal cell carcinoma in an area without obvious arsenic exposure. Toxicol Appl Pharmacol 262 (3): Kanaya S, Ikeya M, Yamamoto K, Moriya T, Furuhashi K, Sonoda M, Goto K, Ochi H (2004) Comparison of an oxidative stress biomarker urinary8-hydroxy-2 -deoxyguanosine, between smokers and non-smokers. BioFactors 22 (1-4): Bogdanov MB, Beal MF, McCabe DR, Griffin RM, Matson WR (1999) A carbon column-based liquid chromatography electrochemical approach to routine 8-hydroxy-2'-deoxyguanosine measurements in urine and other biologic matrices: a one-year evaluation of methods. Free Radic Biol Med 27 (5-6): Benowitz N, Goniewicz ML, Eisner MD, Lazcano-Ponce E, Zielinska-Danch W, Koszowski B, Sobczak A, Havel C, Jacob P, 3rd (2010) Urine cotinine underestimates exposure to the tobacco-derived lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in passive compared with active smokers. Cancer Epidemiol Biomarkers Prev 19 (11): Hu CW, Hsu YW, Chen JL, Tam LM, Chao MR (2014) Direct analysis of tobacco-specific nitrosamine NNK and its metabolite NNAL in human urine by LC-MS/MS: evidence of linkage to methylated DNA lesions. Arch Toxicol 88 (2): de Weerd S, Thomas CMG, Kuster JETG, Cikot RJLM, Steegers EAP (2002) 5

6 Variation of Serum and Urine Cotinine in Passive and Active Smokers and Applicability in Preconceptional Smoking Cessation Counseling. Environ Res 90 (2): Al Ali R, Rastam S, Ibrahim I, Bazzi A, Fayad S, Shihadeh AL, Zaatari GS, Maziak W (2015) A comparative study of systemic carcinogen exposure in waterpipe smokers, cigarette smokers and non-smokers. Tob Control 24 (2): Yuan JM, Gao YT, Wang R, Chen M, Carmella SG, Hecht SS (2012) Urinary levels of volatile organic carcinogen and toxicant biomarkers in relation to lung cancer development in smokers. Carcinogenesis 33 (4): Lee HL, Hsueh YM, Chung CJ, Pu YS, Chang LW, Hsieh DPH, Liou SH, Lin P (2008) Correlation between the Urine Profile of 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanone Metabolites and N7-Methylguanine in Urothelial Carcinoma Patients. Cancer Epidemiology Biomarkers & Prevention 17 (12): Chung CJ, Pu YS, Shiue HS, Lee HL, Lin P, Yang HY, Su CT, Hsueh YM (2013) 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism-related enzymes gene polymorphisms, NNK metabolites levels and urothelial carcinoma. Toxicol Lett 216 (1):

Variation of Urinary 8-Hydroxy-deoxyguanosine in Patients during Radio-therapeutic Course

Variation of Urinary 8-Hydroxy-deoxyguanosine in Patients during Radio-therapeutic Course Hsueh-Hsuan Liu 1, Ing-Jane Chen 1, Chiuan-Chian Chiou 2 1 Dept. of Health Physics, Institute of Nuclear Energy