HPLC ile vitamin A ve E'nin floresans dedektör ve ters faz kolon aracılığında miktar tayini ve kolon performansının ölçümü

Bu çalışmada Hypersil ODS kolonun ve floresans dedektörün Vitamin A ve E'nin tayini için kullanılması ve performanslarını ölçümü amaçlanmıştır. Vitamin A ve E'nin floresans dedektörle aynı ortamda farklı programla (vitamin A: Ex:340nm-Em:460, Vitamin E: Ex:298-Em:350) okunabildiği ve bu amaç için Hypersil ODS ters faz kolonunun kullanılabileceği açıklanmıştır. Kolon; içerdiği dolgu (5 um, RCM 100 RP18. reversed Phase) maddesi açısından hassas ve yüksek ayırıcı özelliğe sahiptir. Vitaminler; özellikle yağda çözünenler, ekstraksiyon ve miktar tayini açısından zorluklar arz etmektedirler. Hayvancılık, gıda. ilaç ve kimya sektöründe 70'li yıllardan günümüze kadar kullanılan HPLC, kolon kapasitesi ve kolon dolgu maddesinden kaynaklanan'özelliklerinden dolayı zorluklar gösteren bir sistemdir. Kolonların teknik verileri; uygulama esnasındaki kullanılan taşıyıcı sıvının ve tespit edilen maddenin özelliğine bağlı olarak değişim göstermektedir.Bu çalışmada akış hızı ile yapılan performans tayininde ve program çalışmasında Vitaminlerin kromatogramlan vemeydana getirdikleri alanlar tespit edilmiştir. Bu sonuçlarla konsantrasyon tayini yapılmıştır.

Quantitative determination of vitamin A and E through florecence dedector and reversed phase column and measurement of column performance by HPLC

The aim of this study is the using and performance controling of Hypersil ODS Column and Florescence dedektor with HPLC. With the same phase and with different program (vitamin A: Ex: 340nm-Emx 460, Vitamin E: Ex: 298-Em: 350). Vitamin A and E were dedected and to this target Hypersil ODS reversed phase column was found useful in this work. With its filling material (5 um, RCM 100 RP18, reversed Phase) column is very sensitive and has a speciall highseparator peculiarity. Vitamins, especially latsoluble vitamins; has more difficulties to extract and to determine. HPLC is used since seventies in the animal, food ,medical and chemical industry and has more difficulties with its filling material and capacity of colons. Technical datum of.column is very variable with flow liquid, and dedected material. In this work we found timing of vitamins and their area and performans of column with flow rate of HPLC. With this result consanlrations can be determined.

PDF

___

1.Barbas, C, Mario, C, Bartolome, B., Marta, V., Emilio, İL: Simultaneous determination of vitamins A and E in rat tissues by high-performance liquid chromatography. Journal of Chromatography A, 1997; 778: 415-420.
2.Brown, K., Reid, A., White, T., Henderson, T., Hukin, S., Johnstone, C, Glen, A.: Vitamin E, lipids, and lipid peroxidation products in tardive dysinesa. Society of Biological Psychiatry, 1998; 43: 863-867.
3.Crissey, S. D., Patricia McGill., Ann-Marie, S.: Influence of dietary vitamins A and E serum a- and y-tocopherols, retinol retinyl palmitate and carotenoid concentrations in Humboldt penguins (Spheniscus humboldti). Comperative Biochemistry and Physiology, Part A, 1998; 121:333-339.
4.Crissey, S. D., Randall, W.: Serum a- and y-tocopherols and retinol, retinyl palmitat, and carotenoid concentrations in captive and free-ranging bottlenose dolphins (Tursiops truncatus). Comperative Biochemistry and Physiology, Part B, 1999; 124: 391-396.
5.Furusawa, N.: Rapid high-performance liquid chromatograhic identification/quantification of total vitamin C in fruit drinks. Food Control, 2001; 12: 27-29.
6.Gimeno, E., Catellote, A. I., Lamuela-Raventos, R. M., de la Torre, M. C, Lopez- Sabater, M. C: Rapid determination of vitamin E in vegetables oils by reversed-phase high- performance liquid chromatography. Journal of Chromatography A, 2000; 881: 251-254.
7.Hiroshi, L: Determination of tocopherol acetate in emulsified nutritional supplements by solid-phase extraction and hihg-performance liquid chromatography with fluorecence detection. Journal of Chromatography A, 2000; 881: 243-249.
8.Melchert, H. U., Pabel, E.: Quantitative determination of a-,(3-,y- and 8-tocopherols in human serum by high-performance liquid chromatography and gas chromatography- mass spectrometry as trimethylsilyl derivatives with a two-step sample preparation. Journal of Chromatography A, 2000; 896: 209-215.
9.Moreno, P., Salvado, V.: Determination of eight water -and fat-soluble vitamins in multi- vitamin pharmaceutical formulations by high-performance liquid chromatography. Journal of Chromatography A, 2000; 870: 207-215.
10.Rizzo, C, Dionisi-Vici, C, D'Ippoliti, M., Fina, F., Sabetta, G., Federic, G.: A simple and rapid HPLC method for simultaneous determination of plasma 7-dehydrocholesterol and vitamin E: its application in Smith-Lemli-Opitz patients. Clinica Chimica Acta, 2000; 291:97-102.
11.Sirimanne, S. R., Patterson, D. G., Ma, L., Justice. J. B.: Application of cloud-point extraction-reversed-phase high performance liquid chromatography: A preliminary study of extraction and quantification of vitamins and E in human serum and whole blood, Journal of Chromatography B, 1998; 716: 129-137.
12.Tavridou, A., Unwin, N. C, Laker, M. F., White, George, K., Alberti, M. M.: Serum concentrations of vitamins A and E in impaired glucose tolerance. Clinica Chimica Acta, 1997; 266: 129-140.
13.Vaananen, P. S., Ollilainen, V., Mattila, P., Klehikoinen, R., Salmela-Mölsa, E., Piironen, V.: Simultaneous HPLC analysis of fat-soluble vitamins in selected animal products after small-scale extraction. Food Chemistry, 2000; 7: 535-543.