OXIDATION KINETICS AND QUANTIFICATION METHOD OF CHOLESTEROL USING CHOLESTEROL OXIDASE ENZYME CATALYST

https://doi.org/10.22146/ijc.21486

Iip Izul Falah(1*), Ritmaleni Ritmaleni(2), M. Utoro Yahya(3)

(1) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(2) Faculty of Pharmacy, Universitas Gadjah Mada, Bulaksumur, Yogyakarta
(3) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(*) Corresponding Author

Abstract


In view of health, cholesterol is believed as one of many sources can raise several diseases. Hence, both of research in quantification and developing simple, rapid and accurate analysis method of cholesterol in a sample is very important. Aim of this research was to investigate cholesterol oxidation kinetics and its quantification method based on oxidation of cholesterol and formation complex compound of hexathiocyanato ferat(III), {Fe(SCN)6}-3. The kinetics analysis and quantification, involved cholesterol oxidation in 0.1 M and pH 7.0 phosphate buffer solution to produce cholest-4-en-3-one and hydrogen peroxide, in the presence of cholesterol oxidase enzyme. The formed hydrogen peroxide was used to oxidize iron(II) ion, which was reacted furthermore with thiocyanate ion to raise the red-brown complex compound. Results of the study showed that the complex was stable at 10-120 min since the reaction was started, with maximum wavelength of 530-540 nm. While the kinetics analysis gave first order oxidation reaction with a reaction rate constant, kapp = 5.22 x 10-2 min-1. Based on this kinetics data, cholesterol analysis method could be developed i.e. by oxidizing cholesterol within 1.5 h using cholesterol oxidase as a catalyst, and then reacted with Fe2+, in a solution containing thiocyanate ion. Absorbencies of solutions of the complex compound, measured at wavelength of 535 nm, were linearly proportional to their cholesterol concentrations, in the range of 50-450 ppm.


Keywords


cholesterol; quantification; kinetics; hexathiocyanato ferat(III)

Full Text:

Full Text Pdf


References

[1] Glasziou, P.P., Irwig, L., Heritier, S., Simes, R.J., and Tonkin, A., 2008, Ann. Intern. Med., 148, 9, 656-661.

[2] Naughton, M.J., Luepker, R.V., and Stricland, D., 1990, J. Am. Med. Assoc., 263, 9, 1213-1217.

[3] Hwang, B-S., Wang, J-T., and Choong, Y-M., 2003, J. Food Compos. Anal., 16, 2, 169-178.

[4] Hurst, W.J., Aleo, M.D., and Martin Jr., R.A., 1983, J. Dairy Sci., 66, 2192-2194.

[5] Daneshfar, A., Khezeli, T., and Lotfi, H.J., 2009, J. Chromatogr. B, 877, 4, 456-460.

[6] Okpuzor, J., Okochi, V.I., Ogbunugafor, H.A., Ogbonnia, S., Fagbayi, T., and Obidiegwu, C., 2009, Pak. J. Nutr., 8, 1, 57-62.

[7] Dinh, T.T.N., Blanton Jr., J.R., Brooks, J.C., Miller, M.F., and Thompson, L.D., 2008, J. Food Compos. Anal., 21, 4, 306-314.

[8] Osman, H., and Chin, Y.K., 2006, Malay. J. Anal. Sci., 10, 2, 205-210.

[9] Palmgren, J.J., Toyras, A., Mauriala, T., Monkkonen, J., and Auriola, S., 2005, J. Chromatogr. B, 821, 2, 144-152.

[10] Xu, X.H., Li, R.K., Chen, J., Chen, P., Ling, X.Y., and Rao, P.F., 2002, J. Chromatogr. B, 768, 2, 369-373.

[11] Sion, B., Grizard, G., and Boucher, D., 2001, J. Chromatogr. A, 935, 1-2, 259-265.

[12] Goodhue, C.T., Risley, H.A., and Snoke, R.E., 1975, Method and Composition for Blood Serum Cholesterol Analysis, US Patent, No. 3,884,764.

[13] Allain, C.C., Poon, L.S., Chan, C.S.G., Richmond, W., and Fu, P.C., 1974, Clin. Chem., 20, 4, 470-475.

[14] Raheb, J., Lashgarian, H.E., Memari, B., Hajipour, M.J., and Haghbeen, K., 2009, Inter. J. Biotechnol. Biochem., December 01, 2009, Accessed February 19, 2010, http://www.thefreelibrary.com/Identification,

[15] Anonym, 2009, A Fast Enzymatic Assay for Plasma Cholesterol Using Cholesterol Oxidase, Technical Bulletin: Cholesterol Analysis, Bulletin ref. TB018812F, Accessed February 19, 2010, http://www.analox.com/tbcho018812f.htm.

[16] Bhatia, D., Suman, and Pundir, C.S., 2005, Ind. J. Biotechnol., 4, 471-475.

[17] Kouassi, G.K., Irudayaraj, J., and McCarty, G., 2005, J. Nanobiotechnol., 3,1.

[18] Moreau, R.A., Powell, M.J., and Kevin, B., 2003, J. Agric. Food Chem., 51, 23, 6663-6667.

[19] Kayamori, Y., Hatsuyama, H., Tsujioka, T., Nasu, M., and Katayama, Y., 1999, Clin. Chem., 45, 12, 2158-2163.

[20] Vasudevan, P.T., and Zhou, T., 1997, Biotechnol. Bioeng., 53, 4, 391-396.

[21] Schmarr, H-G., Gross, H.B., and Shibamoto, T., 1996, J. Agric. Food Chem., 44, 2, 512-517.

[22] Olafsdottir, E., and Gudmundsson, T.V., 1993, Upsala J. Med. Sci., 98, 401-404.

[23] Zang, L-Y., Cosma, G., Gardner, H., Shi, X., Castranova, V., and Valiyathan, V., 2000, Am. J. Physiol., 279, 4, 954-960.

[24] Bagheri, H., Gholami, A., and Najafi, A., 2000, Anal. Chim. Acta, 424, 2, 233-242.

[25] Riganakos, K.A., and Veltsistas, P.G., 2003, Food Chem., 82, 4, 637-643.

[26] Basavaiah, K., and Swamy, J.M., 2001, II Farmaco, 56, 8, 578-585.

[27] Deglampietro, P., Peheim, E., Drew, D., Graf, H., and Colombo, J.P., 1987, J. Clin. Chem. Clin. Biochem., 25, 711-717.

[28] Alekssandrovskii, I., 1987, Biokhimiia, 52, 10, 1696-1703.

[29] Vasudevan, P.T., and Zhou, T., 1996, App. Biochem. Biotechnol., 60, 1, 63-72.

[30] Copper, C.L., and Koubek, E., 1999, Inorg. Chim. Acta, 288, 2, 229-232.

[31] Wong, G.T.F., and Zang, L-S., 2008, Marine Chem., 111, 1-2, 22-29.



DOI: https://doi.org/10.22146/ijc.21486

Article Metrics

Abstract views : 1687 | views : 1277


Copyright (c) 2010 Indonesian Journal of Chemistry

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

 

Indonesian Journal of Chemistry (ISSN 1411-9420 /e-ISSN 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

Web Analytics View The Statistics of Indones. J. Chem.