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
Fig. S1 Representative MRM chromatograms of LC-MS/MS analysis for total NNAL in human urine. The concentration of NNAL was 0.081 ng ml -1, and the concentration of IS was 2 ng ml -1 in the human urine 2
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 (82-1200) Smoker (n = 3) [8] On-line SPE LC-MS/MS 638.30 ± 439.57 Smoker (n = 40) Our study Total NNAL HPLC-MS/MS 0.22 (0.13-0.39) Smoker (n = 130) [6] HPLC-MS/MS 0.17 ± 0.14 Smoker (n = 101) [7] HPLC-MS/MS 0.04 (0.03-0.05) 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
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 ± 0.02 1.33 0.04 ± 0.03 1.75 Smoker a (n=40) 0.04 ± 0.02 0.07 ± 0.04 Al Ali R et al. [9] Non-smoker a (n=28) 0.0006 14.8 0.0036 10.86 Smoker a (n=23) 0.0089 0.0391 Hu et al. [7] Non-smoker a (n=40) - b - b Smoker a (n=101) 0.10 ± 0.09 0.17 ± 0.14 Lee et al. [11] Non-smoker a (n=69) 0.48 ± 0.10 2.20 1.49 ± 0.28 2.12 Smoker a (n=57) 1.06 ± 0.31 3.16 ± 0.99 Chung et al. [12] Non-smoker a (n=81) 0.14 ± 0.03 0.93 0.40 ± 0.07 1.03 Smoker a (n=22) 0.13 ± 0.03 0.41±0.09 a : mean ± SD, b : not mention, c: the concentration of smoker / the concentration of non-smoker. 4
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