Optimized Method of High Performance Liquid Chromatography using Octyl Silica Fully Endcapped Residual Silanol Column on Separation of Cotinine and 3-Hydroxycotinine in Urine Sample

  • CHRISTINE PATRAMURTI UNIVERSITAS PANCASILA
  • SUDIBYO MARTONO UNIVERSITAS SANATA DHARMA
  • SUGIYANTO SUGIYANTO UNIVERSITAS SANATA DHARMA
  • ARIEF NURROCHMAD UNIVERSITAS SANATA DHARMA

Abstract

Cotinine (COT) and 3-hydroxycotinine (3-HCOT) are nicotine metabolite excreted in urine. Mediated by the enzyme cytochrome P450 2A6 (CYP 2A6), nicotine will be metabolized to COT and 3-HCOT. The activity of CYP 2A6 can be predicted from the ratio 3-HCOT to the COT, therefore the ratio of 3-HCOT and COT can be used as phenotyping and polymorphism studies of the enzyme. In this study, isolation COT and 3-HCOT of urine samples was carried out by liquid-liquid back extraction. Simultaneous analysis of COT and 3-HCOT using High Performance liquid Chromatography (HPLC) was performed by a reversed-phase octyl silica column (C8; Shimadzu 250 × 4.6 mm, 5 μm) fully endcapped residual silanol. The internal standard solution (SI) was acetanilide. The mobile phase which separate COT, 3-HCOT and SI was methanol : ammonium acetate 5 mM (50:50) at a flow rate 0.8 mL/min. Retention time (tR) of the three compounds was less than 10 minutes, with peak tailing factor (tf) was less than 2. The resolution (Rs) 3-HCOT to COT was 2.67, while the Rs COT to SI was 8.836.

References

1. Benowitz NL and Jacob P. Metabolism of nicotine to cotinine studied by a dual stable isotope method. Clin Pharmacol Ther. 1994. 56:483-93.
2. Jacob P, Yu L, Duan M, Ramos L, Yturralde O, Benowitz NL. Determination of the nicotine metabolites cotinine and trans-3-hydroxycotinine in biologic fluids of smokers and non-smokers using liquid chromatography-tandem mass spectrometry: Biomarkers for tobacco smoke exposure and for phenotyping cytochrome P450 2A6 activity. J Chromatogr B. 2011. 879:267-76.
3. Tuomi T, Johnsson T, and Reijula K. Analysis of nicotine, 3-hydroxycotinine, cotinine and caffeine in urine of passive smokers by HPLC-tandem mass spectrometry. Clin Chem. 1999. 45:2164-72.
4. Benowitz NL, Pomerleau OF, Pomerleau CS, Jacob P. Nicotine metabolite ratio as a predictor of cigarette consumption. Nicotine Tob Res. 2003. 5:621-4.
5. Derby KS, Cuthrell K, and Caberto C. Nicotine metabolism in three ethnic/racial groups with different risks of lung cancer. Cancer Epidemiol. Biomarkers Prev. 2008. 17:3526-35.
6. Sellers EM and Tyndale RF. Mimicking gene defects to treat drug dependence. Ann NY Acad Sci. 2000. 909:233-46.
7. Kubota T, Nakajima-Taniguchi C, Fukuda T, Funamoto M, Maeda M, Tange1 E, et al. CYP2A6 polymorphisms are associated with nicotine dependence and influence withdrawal symptoms in smoking cessation. The Pharmacogenomics J. 2006. 6:115-9.
8. Massadeh AM, Gharaibeh AA, and Omari KW. A single-step extraction method for the determination of nicotine and cotinine in Jordanian smokers’ blood and urine samples by RP-HPLC and GC–MS. J Chromatogr Sci. 2009. 47:170-7.
9. Rabbaa-Khabbaz L, Daoud RA, and Karam-Sarkis D. A simple, sensitive, and rapid method for the determination of cotinine in urine by high-performance liquid chromatography with UV detection. J Chromatogr Sci. 2006. 44:535-8.
10. Wen YH, Yang PS, and Wu SS. Determination of cotinine in human urine by high-performance liquid chromatography. JFDA. 2009. 17(5):357-62.
11. Badran E, Salhab AS, and Al-Jaghbir M. Infant exposure to environmental tobacco smoke: Jordan University Hospital-based study. Eastern Mediterranean Health J. 2009. 15(1):39-46.
12. Man CN, Gamb L, Ismail S, Lajis R, Awang R. Simple, rapid and sensitive assay method for simultaneous quantification of urinary nicotine and cotinine using gas chromatography–mass spectrometry. J Chromatogr B. 2006. 844:322-7.
13. Shin H, Kim J, Shin Y, Jee SH. Sensitive and simple method for the determination of nicotine and cotinine in human urine, plasma and saliva by gas chromatography–mass spectrometry. J Chromatogr B. 2002. 769:177–83.
14. Tyrpien K, Wielkoszyn´skia T, Janoszkaa B, Dobosza C, Bodzek D, Steplewski Z. Application of liquid separation techniques to the determination of the main urinary nicotine metabolites. J Chromatogr A. 2000. 870:29–38.
15. Bao Z, He X, Ding X, Prabhu S, and Hong J. Metabolism of nicotine and cotinine by human cytochrome P450 2A13. DMD. 2005. 33:258-61.
16. Doctor PB, Gokani VN, Kulkarni PK, Parikh JR, Saiyed HN. Determination of nicotine and cotinine in tobacco harvesters’ urine by solid-phase extraction and liquid chromatography. J Chromatogr B. 2004. 802:323-8.
17. Baumanna F, Regenthala R, Burgos-Guerrerob IL, Hegerlb U, Preiss R. Determination of nicotine and cotinine in human serum by means of LC/MS. J Chromatogr B. 2010. 878:107-11.
18. Meger M, Meger-Kossien I, Schuler-Metz A, Janket D, Scherer G. Simultaneous determination of nicotine and eight nicotine metabolites in urine of smokers using liquid chromatography–tandem mass spectrometry. J Chromatogr B. 2002. 778:251-61.
19. Moyer TP, Charlson JR, Enger RJ, Dale LC, Ebbert JO, Schroeder DR, and Hurt RD. Simultaneous analysis of nicotine, nicotine metabolites, and tobacco alkaloids in serum or urine by tandem mass spectrometry, with clinically relevant metabolic profiles. Clin Chem. 2002. 48:1460-71.
20. Shakleya DM, Huestis MA. Simultaneous and sensitive measurement of nicotine, cotinine, trans-3-hydroxycotinine and norcotinine in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B. 2009. 877:3537-42.
21. Page-Sharp M, Hale TW, Hackett LP, Kristensen JH, Ilett KF. Measurement of nicotine and cotinine in human milk by high-performance liquid chromatography with ultraviolet absorbance detection. J Chromatogr B. 2003. 796:173–80.
22. Snyder LR, Kirkland JJ, Dolan JW. Introduction to modern liquid chromatography. 3th ed. New Jersey: John Wiley & Sons, Inc.; 2010. 51-9, 222, 327.
23. Bhalala O. Detection of cotinine in blood plasma by HPLC MS/MS. MURJ. 2003. 8:45-50.
Published
2014-04-30
How to Cite
PATRAMURTI, CHRISTINE et al. Optimized Method of High Performance Liquid Chromatography using Octyl Silica Fully Endcapped Residual Silanol Column on Separation of Cotinine and 3-Hydroxycotinine in Urine Sample. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 12, n. 1, p. 110-116, apr. 2014. ISSN 2614-6495. Available at: <http://jifi.farmasi.univpancasila.ac.id/index.php/jifi/article/view/195>. Date accessed: 22 july 2024.
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Vol 12 No 1 (2014): JIFI
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