- P-ISSN 1225-0163
- E-ISSN 2288-8985
A simultaneous determination method for cholesterol, lanosterol and desmosterol was developed using gas chromatography/mass spectrometry. Urine was enzymatically hydrolyzed with β-glucuronidase/arylsulfatase. Samples were prepared using extractions with a mixture of ethyl acetate-hexane (2:3, v/v), followed by derivatization with a mixture of MSTFA/TMSI/TMCS (100:2:5 v/v/v). All analyses were performed using GC/ MS in selective ion monitoring mode. Good linearities (r2=0.998~0.999) in calibration curve and a satisfactory recovery (80.0%~113%) were achieved. Accuracy and precision values within ±15% in the concentration range of 5 to 200 ng/mL were also observed for all compounds. The developed method was applied to pravastatintreated (70 and 250 mg/kg/day for 7 days, oral) hyperlipidemia rats. Those sterols were significantly lower in drug-treated rats compared to the controls, which justifies the drug efficacy. Therefore, these results indicate that the developed method was successfully applied to examine statin drug efficacy with urine sample.
1. C. S. J. W. Briche, D. Carter and K. S.Webb, Rapid. Commun. Mass Spectrom., 16, 848-853(2002).
2. J. K. Liao, Hypertension, 43, 1171-1172(2004).
3. K. S. Jain, M. K. Kathiravan, R. S. Somania and C. J. Shishooc, Bioorg. Med. Chem., 15, 4674-4699(2007).
4. K. Fassbender, D. Lütjohann, M. G. Dik, M. Bremmer, J. König, S. Walter, Y. Liu, M. Letièmbre, K. von Bergmann and C. Jonker, Atherosclerosis, 196, 283– 288(2008).
5. T. A. Miettinen, Life Sci., 8, 713-721(1969).
6. T. A. Miettinen, Ann. Clin. Res., 2, 300-320(1970).
7. R.A. Rajaratnam, H. Gylling and T. A. Miettinen, J. Am. Coll. Cardiol., 35, 1185-1191(2000).
8. T. A. Miettinen, T. E. Strandberg, H. Gylling and the Finnish Investigators of the Scandanavian Simvastatin Survival Study Group, Arterioscler. Thromb. Vasc. Biol., 20, 1340-1346(2000).
9. T. A. Miettinen and H. Gylling, Am. J. Cardiol., 96, 40- 46(2005).
10. K. Nagy, A. Jakab, F. Pollreisz, D. Bongiorno, L. Ceraulo, M. R. Averna, D. Noto and K.Vékey, Rapid. Commun. Mass Spectrom., 20, 2433-2440(2006).
11. A. L. Garcýá-Otý´n,M. Cofa´n,M. Junyent, D. Recalde, A. Cenarro, M. Pocovýí , E. Ros and F. Civeira,
13. Y. Aoyama, Y. Yoshida and R. Sato, J. Biol. Chem., 259, 1661-1666(1984).
14. D. Lütjohann, A. Brzezinka, E. Barth, D. Abramowski, M. Staufenbiel, K. von Bergmann, K. Beyreuther, G. Multhaup and T.A. Bayer, J. Lipid Res., 43, 1078- 1085(2002).
15. F. Chevy, L. Humbert and C. Wolf, Prenat Diagn., 25,1000-1006(2005).
16. N. Yamaga, Y. Ogura, M. A. Islam, H. Matsuyama, M. Yamawaki, M. Kai and K. Yamada, Yonago Acta medica., 45, 27-3(2002).
17. A. Kuksis, J. Chromatogr. A, 935, 203-236(2001).
18. H. Shan, J. Pang, S. Li, T.B. Chiang, W.K. Wilson, G.J. Jr. Schroepfer, Steroids, 68, 221-233(2003).
19. E. de Rijke, B. Ruisch, J. Bakker, J. Visser, J. Leenen, S. Haiber, A. de Klerk, C. Winkel and T. König, J. Agric. Food Chem., 55, 6417-6423(2007).
20. D. Huster, H.A. Scheidt, K. Arnold, A. Herrmann and P. Müllerz Biophys. J., 88, 1838-1844(2005).
21. T. Soga, Y. Ohashi, Y. Ueno, H. Naraoka, M. Tomita and T. Nishioka, J. Proteome Res., 2, 488-494(2003).
22. M. Kivilompolo, V. Obùrka and T. Hyötyläinen, Anal. Bioanal. Chem., 388, 881-887(2007).
23. J. H. Herndon and M. D. Siperstein, Circ. Res., 12, 228- 234(1963).
24. E. Hansbury and T. J. Scallen, J. Lipid Res., 19, 742- 746(1978).
25. C. Domenõ, B. Ruiz, C. Nerín, Anal. Bioanal. Chem., 381, 1576-1583(2005).
26. J. C. Penteado, R. E. Bruns, L. R. Franco de Carvalho, Anal. Chim. Acta, 562, 152-157(2006).
27. Y. Tsujita, M. Kuroda, Y. Shimada, K. Tanzawa, M. Arai, I. Y. Tsujita, M. Kuroda, Y. Shimada, K. Tanzawa, M. Arai and I. Fujii, Biochim. Biophys. Acta, 877, 50- 60(1986).
28. H. Mabuchi, T. Haba, R. Tatami, S. Miyamoto, Y. Sakai, T. Wakasugi, A. Watanabe, J. Koizumi and R. Takeda, N. Engl. J. Med., 305, 478-482(1981).
29. N. Nakaya, Y. Homma, H. Tamachi, H. Shigematsu, Y. Hata, Y. Goto, J. Am. Med. Assoc., 257, 3088-3093 (1987).
30. J. Y. Moon, H. J. Jung, M. H. Moon, B. C. Chung and M. H. Choi, J. Am. Soc. Mass Spectrom. doi: 10.1016/ j.jasms.2009.04.020.
31. J. P. Kassirer, N. Engl. J. Med., 285, 385-389(1971).
32. R. Causon, J. Chromatogr. B, 689, 175-180(1997).