Comparision of Preparation Methods for Water Soluble Vitamin Analysis in Foods by Reversed-Phase High Performance Liquid Chromatography

역상 고속 액체 크로마토그래피에 의한 식품 중 수용성 비타민 분석을 위한 전처리법의 비교

  • Published : 2002.04.01


Owing to a need for simple extraction and purification for analysis of water soluble vitamins in food samples by RP-HPLC with UV-detector, the methods of bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were employed. The recoveries of standard water soluble vitamins by the bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were significantly high compared to AOAC methods in most of vitamins. The contents of pyridoxal determined with protest in the pork was similar, but in the bromelain hydrolysis and AOAC method, was high compared to the results of reference. The niacinamide, thiamin and riboflavin determined with bromelain and protease hydrolysis showed similar values to the results of references. In the potato, pyridoxamine was detected in the AOAC method, which was not detected in the bromelain and protease hydrolysis methods. Pyridoxal contents in the protease hydrolysis and AOAC methods were very similar to the results of references. The recoveries of fortified standard vitamins in food samples were significantly high and accurate compared to those of AOAC methods. The extraction and purification with $C_{18}$ Sep-Pak solid extractor might be considered superior method for the determination of water soluble vitamins in food samples.


  1. Augustin, J. Simultaneous determlnation of thiamine and ribofla-vin in foods by liquid chromatography. J. A.O.A.C. 67: 1012-1015(1984)
  2. Van de Weerdhof, T., Wiersum, M.L. and Reissenweber, H.Application of liquid chromatography in food analysis. J. Chro-matogr. 83: 455-460 (1973)
  3. Williams, R.C., Baker, D.R. and Schmit, J.A. Analysis of water soluble vitamins by high speed ion exchange chromatography. J.Chromatogr. Sci. 11: 618-624 (1973)
  4. A.O.A.C. Offcial Methods of Analysis. 16th ed. Association of Official Analytical Chemists, Washington, DC, USA (1995)
  5. Gregory, J.F. and Kirk, J.R. Assessment of storage effects on vita-min B6 stability and bioavailability in dehydrated food systems.J. Food Sci. 43: 1801-1808 (1978)
  6. Wong, F.F. Analysis of vitamin B6 in extractives of food materi-als by high-performance liquid chromatography. J. Agric. Food Chem. 26: 1444-1446 (1978)
  7. Ohta, H., Baba, T. and Suzuki, Y. High-performance liquid chromatographic analysis of thiamin in rice flour with fluorimetric post-column dervatization. J. Chromatogr. 284: 281-284 (1984)
  8. Ang, C.Y. and Mosley, F.A. Determination of thiamin and ribofla- vin in meat products by high-pressure liquid chromatography. J.Agric. Food Chem. 28: 483-486 (1980)
  9. Clifford, C.K. and Clifford, A.J. High pressure liquid chromato-graphic analysis of food for folates J. A.O.A.C. 60: 1248-1251(1977)
  10. Dickey, L.E. Composition of foods, raw, Processed, Prepared, sup-plement, No. 8, p. 330. United States Department of Aghculture,Human nutridon information survice, Agriculture Handbook (1990)
  11. Freed, M. Method of vitamin analysis. Interscience, New York, USA(1966)
  12. Yasumoto, K. Tadera, K., Tsuji, H. and Mitsuda, H. Semi-auto-mated system for analysis of vitamin B6 complex by ionexchange column chromatography. J. Nutr. Sci. Vitamol. 21: 117-127 (1975)
  13. Hoppner, K. and Lampi, B. The determination of folic acid (pteroylmonoglutamic acid) in fortified products by reversed phase high pressure liquid chromatography. J. Liquid Chromatogr.5: 953-966 (1982)
  14. De Leenheer, A.P., Lambert, W.E. and De Ruyter, M.G. Modern Chromatographic Analysis of Vitamins. Marcel Dekker, New York. (1985)
  15. Reingold, R.N. and Picciano, M.F. Two improved high-perfor-mance liquid chromatographic separation. J. Chromatogr. 234:171-179(1982)
  16. Lambert, W.E. and Leenheer, A.D. Quantitative determination of water soluble vitamins using HPLC. pp. 341. In: Food Analysis by HPLC. Nollet, L.M.L. (ed.). Marcel Dekker Inc, New York, USA (1992)
  17. Schiefer, G.W., Wheeler, G.P. and Cimino, C.M. Determination of folic acid in commercial diets by anion-exchange solid-phase extraction and subsequent reversed-phase HPLC. J. Liquid Chro-matogr. 7: 2659-2669 (1984)
  18. Friednch, W. Vitamins. Walter de Gruyter, New York, USA(1988)
  19. Wimalasiri, P. and Wills, R.B.H. Simultaneous analysis of thiamin and riboflavin in foods by high performance liquid chromatogra-phy. J. Chromatogr. 318: 412-416 (1985)
  20. Van Nieker, P.J. HPLC in food analysis, pp. 133. MaCrae, R. (Ed.), Academic Press, New York (1988)
  21. Vanderslice, J.T., Brownlee, S.R. and Cortissoz, M.E. Liquid chromatographic determination of vivamin B6 in foods. J.A.O.A.C.67:999-1007(1984)
  22. Hilker, D.M. and Clifford, A.J. Thiamin analysis and separation of thiamin phosphate esters by high-performance liquid chroma-tography. J. Chromatogr.231: 433-438 (1982)
  23. Osborne, D.R. and Voogt, P. The Analysis of Nntrients in Food. Academic Press, New York, USA (1978)
  24. Fellman, J.K., Artz, W.E., Tassinari, P.D., Cole, C.L. and Augus-tin, J. Simultaneous determination of thiamin and riboflavin in selected foods by high-performance liquid chromatography. J.Food Sci. 47: 2048-2067 (1982)
  25. Kral, K. Bestimmung von nicotins ure in fruchts ften durch HPLC mit elektrochemischer detektion an einer station renquecksilbertropfelektrode. Fresenius Z. Anal. Chem. 314: 479-482(1983)
  26. Defibaugh, P.W. Evaluation of selected enzymes for thiamin determination, J. A.O.A.C. 70(3): 514-517 (1987)
  27. Kamman, J.K., Labuza, T.P. and Warthesen, J.J. Thiamin and riboflavin analysis by high performance liquid chromatography. J.Food Sci.45: 1497-1504(1980)
  28. Gregory, J.F., Sartain, D.B. and Day, B.D. Fluorometric determi-nation of folacin in biological materials using high-performance liquid chromatography. J. Nutr. 114: 341-353 (1984)
  29. Coffen, D.L. Kirk-Othmer Encyclopedia of Chemical Technology.Wiley, New York, USA (1984)
  30. Ayi, B.K., Yuhas, D.A., Moffet, K.S., Joyce, D.M. and Deangelis,N.J. Liquid chromatographic determination of thiamin in infant fomula products by using ultraviolet detection. J. A.O.A.C. 68(6):1087-1092(1985)
  31. Holt, D.L., Wheling, R.L. and Zeece, M.G. Determination of native folates in milk and other diarary products by high perfor- mance liquid chromatography. J. Chromatogr. 449: 271-280 (1988)
  32. Tyler, T.A. and Shrago, R.R. Determination of niacin in cereal samples by HPLC. J. Liquid Chromatogr. 3: 269-277(1980)
  33. MacBride, D.E. and Wyatt, C.J. Evaluation of a method AOACdetermination for thiamine and riboflavin in foods. J. Food Sci.48: 748-750 (1983)