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[Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine by Hydrophilic Chromatography-tandem Mass Spectrometry with Isotope Dilution].
Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Jan; 51(1):74-80.SD

Abstract

OBJECTIVE

To develop an assay for determination of 8-oxo-2'-deoxyguanosine and cotinine in human urine by hydrophilic chromatography tandem mass spectrometry (HILIC-MS/MS) with isotope dilution.

METHODS

The urine supernatant was 1∶5 diluted with 3 mmol/L ammonium formate aqueous solution containing ¹⁵N ₅-8-OHdG and D ₃-cotinine as internal standard. After being filtered through a 0.22 μm water filter, the sample solution was injected into ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Separation was performed on ACQUITY UPLC® BEH HILIC column (50 mm×3.0 mm, 1.7 μm) with isocratic elution (A∶B=10∶90) at 40 ℃. The mobile phase was composed with acetonitrile (B) and 3 mmol/L ammonium formate water soulution (A). The flow rate was 0.3 mL/min. Positive ion scan-multiple reaction monitoring (MRM) mode were used for monitoring and internal standard curves were applied for quantification.

RESULTS

Good linearity was obtained under the optimal conditions. Detection limits for 8-OHdG and cotinine were 0.064 µg/L and 0.035 µg/L respectively, the quantitation limits were 0.21 µg/L and 0.12 µg/L respectively, and the recoveries of the spiked urine samples were 92.6%-102% and 102%-106% respectively. Statistical analysis of 40 urine sample determination results obtained by using the above assay showed that there were significant differences in tobacco smoke exposure and tobacco-specific nitrosamine intake between active and passive smoker ( P<0.05). The concentration of NNAL and cotinine were higher in urine samples of active smoker. Tobacco smoke exposure was positively correlated with tobacco specific nitrosamine intake in both active and passive smokers (the correlation coefficients were 0.487 and 0.786 respectively, P<0.05).

CONCLUSION

We successfully established a simple and fast assay for simultaneously detecting 8-oxo-2'-deoxyguanosine and cotinine in human urine. It was sensitive and accurate for quntification via the calibration by the isotope internal standards, and can meet the needs of batch analysis.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

Department of Laboratory Technology and Science of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.

MeSH

8-Hydroxy-2'-DeoxyguanosineChromatography, High Pressure LiquidCotinineHumansIsotopesTandem Mass SpectrometryUrinalysis

Pub Type(s)

Journal Article

Language

chi

PubMed ID

31950793

Citation

Yang, Ming-Qi, et al. "[Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine By Hydrophilic Chromatography-tandem Mass Spectrometry With Isotope Dilution]." Sichuan Da Xue Xue Bao. Yi Xue Ban = Journal of Sichuan University. Medical Science Edition, vol. 51, no. 1, 2020, pp. 74-80.

Yang MQ, Yuan Y, Ren JW, et al. [Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine by Hydrophilic Chromatography-tandem Mass Spectrometry with Isotope Dilution]. Sichuan Da Xue Xue Bao Yi Xue Ban. 2020;51(1):74-80.

Yang, M. Q., Yuan, Y., Ren, J. W., Zhu, J., Wang, Y., Wang, W. J., & Zou, X. L. (2020). [Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine by Hydrophilic Chromatography-tandem Mass Spectrometry with Isotope Dilution]. Sichuan Da Xue Xue Bao. Yi Xue Ban = Journal of Sichuan University. Medical Science Edition, 51(1), 74-80. https://doi.org/10.12182/20200160102

Yang MQ, et al. [Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine By Hydrophilic Chromatography-tandem Mass Spectrometry With Isotope Dilution]. Sichuan Da Xue Xue Bao Yi Xue Ban. 2020;51(1):74-80. PubMed PMID: 31950793.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - [Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine by Hydrophilic Chromatography-tandem Mass Spectrometry with Isotope Dilution]. AU - Yang,Ming-Qi, AU - Yuan,Yue, AU - Ren,Jian-Wei, AU - Zhu,Jing, AU - Wang,Yan, AU - Wang,Wen-Jia, AU - Zou,Xiao-Li, PY - 2020/1/18/entrez PY - 2020/1/18/pubmed PY - 2020/2/7/medline KW - 8-oxo-2'-deoxyguanosine KW - Cotinine KW - HILIC-MS/MS KW - Isotope dilution KW - Urine SP - 74 EP - 80 JF - Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition JO - Sichuan Da Xue Xue Bao Yi Xue Ban VL - 51 IS - 1 N2 - OBJECTIVE: To develop an assay for determination of 8-oxo-2'-deoxyguanosine and cotinine in human urine by hydrophilic chromatography tandem mass spectrometry (HILIC-MS/MS) with isotope dilution. METHODS: The urine supernatant was 1∶5 diluted with 3 mmol/L ammonium formate aqueous solution containing 15N 5-8-OHdG and D 3-cotinine as internal standard. After being filtered through a 0.22 μm water filter, the sample solution was injected into ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Separation was performed on ACQUITY UPLC® BEH HILIC column (50 mm×3.0 mm, 1.7 μm) with isocratic elution (A∶B=10∶90) at 40 ℃. The mobile phase was composed with acetonitrile (B) and 3 mmol/L ammonium formate water soulution (A). The flow rate was 0.3 mL/min. Positive ion scan-multiple reaction monitoring (MRM) mode were used for monitoring and internal standard curves were applied for quantification. RESULTS: Good linearity was obtained under the optimal conditions. Detection limits for 8-OHdG and cotinine were 0.064 µg/L and 0.035 µg/L respectively, the quantitation limits were 0.21 µg/L and 0.12 µg/L respectively, and the recoveries of the spiked urine samples were 92.6%-102% and 102%-106% respectively. Statistical analysis of 40 urine sample determination results obtained by using the above assay showed that there were significant differences in tobacco smoke exposure and tobacco-specific nitrosamine intake between active and passive smoker ( P<0.05). The concentration of NNAL and cotinine were higher in urine samples of active smoker. Tobacco smoke exposure was positively correlated with tobacco specific nitrosamine intake in both active and passive smokers (the correlation coefficients were 0.487 and 0.786 respectively, P<0.05). CONCLUSION: We successfully established a simple and fast assay for simultaneously detecting 8-oxo-2'-deoxyguanosine and cotinine in human urine. It was sensitive and accurate for quntification via the calibration by the isotope internal standards, and can meet the needs of batch analysis. SN - 1672-173X UR - https://www.unboundmedicine.com/medline/citation/31950793/[Determination_of_8_oxo_2'_deoxyguanosine_and_Cotinine_in_Urine_by_Hydrophilic_Chromatography_tandem_Mass_Spectrometry_with_Isotope_Dilution]_ DB - PRIME DP - Unbound Medicine ER -