Separation and determination of the amino acids by ion exchange column chromatography applying post-column derivatization
Keywords:
ion exchange column chromatography, IEC, high performance liquid chromatography, HPLC, amino acids, determinationAbstract
The most perfect method for the determination of the amino acid composition of pure protein feeds or biological fluids is still the ion exchange column chromatography (IEC). With the help of the lithium buffer system, most of the problems in the field of free amino acid analysis of biological fluids can be solved. At IEC, most contaminants move rapidly through the post-column system and are discarded before the separation of amino acids begins, resulting in better performance. The time required for the sample preparation is minimal compared to pre-column methods, and the detection (with ninhydrin or OPA) is chemically specific for amino acids. A new off-line sample preparation method has been introduced before analysis by an ion-exchange chromatography and integrated amperometric detection, which eliminates carbohydrates from amino acid samples. However, the major problems remained in the hydrolysis of the protein, the de-proteinization of the biological fluids and the partial decomposition of methionine, cysteine and tryptophan during the sample preparation for analysis. At the moment the traditional IEC with post-column ninhydrin derivatization seems to be the best method to analyse both pure proteins and feeds and complex mixtures, but in some comparison the HPLC methods were found to be similar to that of the IEC.
References
Amino acids in animal nutrition. A compendium of recent reviews and reports (2002). Szerk.: Bologa U. L., Degussa 1–558.
Amino acid analyser AAA 400. (2002). User manual INGOS.
Amino acid analysis Handbook and application (1982). LKB Biochrom.
Andersen, T. T. (1994). Practical amino acid analysis. ABRF News, 1–5.
Ashworth, R. B. (1987). J. Assoc. Off. Anal. Chem., 70(2), 248–252. https://doi.org/10.1093/jaoac/70.2.248
Belitz, H. D. Grosch, W. (1999). Food chemistry. Springer, 1–992. https://doi.org/10.1007/978-3-662-07281-3
Le Boucher, J., Charret, C., Coudray-Lucas, C., Giboudeau, J., Cynober, L., (1997). Amino acid determination in biological fluids by antrered ion-exchange chromatography: performance of Hitachi L-8500A. Chinical Chemistry, 43(8), 1421–1428. https://doi.org/10.1093/clinchem/43.8.1421
Cooper, C., Packer, N., Williams, K. (2001). Amino acids protocols. Humana Press, 1–265. https://doi.org/10.1385/1592590470
Davey, J. F., Ersser, R. S. (1990). Amino acid analysis of physiological fluids by high-performance liquid chromatography with phenylisothiocyanate derivatization and comparison with ion-exchange chromatography. J. Chromatography, 528. 9–23. https://doi.org/10.1016/S0378-4347(00)82358-0
Davies, R. L., Bagient, D. R., M. S. Levitt, M. S., Mollah, Y., Rayner, C. J., Frensham, A. B. (1992). Accuracy and precision in amino acid analysis. J. Sci. Food Agric., 59(4), 423–436. https://doi.org/10.1002/jsfa.2740590402
Ding, Y., Yu, H., Mou, S. (2003). Off-line elimination of carbohydrates for amino acid analysis of samples with high carbohydrate content by ion-exchange chromatography. J. Chromatography, 997(1–2), 155–160. https://doi.org/10.1016/S0021-9673(03)00548-X
Friedman, M. (1975). Protein nutrition quality of foods and feeds. Assay methods-biological, biochemical and chemical. Marcel Dekker 1–626.
Grunau, J. A., Swiader, M. (1992). Chromatography of 99 amino acids and other ninhydrin-reactive compounds in the Pickering lithium gradient system. J. Chromatography, 594(1–2), 136–171. https://doi.org/10.1016/0021-9673(92)80326-P
Hanko, V. P., Heckenberg, A., Rohrer, J. S. (2004). J. Biomolecular Tecniques, 15. 317–324.
Hanko, V. P., Rohrer, J. S. (2002). Direct determination of tryptophan using high-performance anion-exchange chromatography with integrated pulsed amperometric detection. Analytical Biochemistry, 308(2), 204–209. https://doi.org/10.1016/S0003-2697(02)00215-4
Holm, D. J., Peck, H. (1998). Analytical Biochemistry. Longman 91–168.
Jandik, P., Cheng, J., Jensen, D., Manz, S., Avdalovic, N. (2001). Simplified in-line sample preparation for amino acid analysis in carbohydrate containing samples. J. Chromatography, 758(2), 189–196. https://doi.org/10.1016/S0378-4347(01)00179-7
Moore, S., Stein, W. H. (1951). Chomatography of amino acids on sulfonated polystyrene resins. J. Biol. Chem., 192. 663–681. https://doi.org/10.1016/S0021-9258(19)77791-6
Moore, S., Stein, W. H. (1963). Chomatographic determination on amino acids by the use of automatic recording equipment. Methods Enzimol., 6. 819–831. https://doi.org/10.1016/0076-6879(63)06257-1
Feed evaluation. Principles and practice. Szerk.: Moughan, P. J., Verstegen, M. W. A., Visser-Reyneveld, M. I. (2000). Wageningen Pers. 1–290.
Parvy, P., Bardet, J., Rabier, D., Gasquet, M., Kamoun, P. (1993). Clin. Chem., 39(9), 1831–1836. https://doi.org/10.1093/clinchem/39.9.1831
Pickering, M. V. (1981). U.S. Patent 4. 274. 833.
Pickering, M. V. (1989). Ion-exchange chromatography of free amino acids. LC-GC 6. 484–490.
Ravindran, G., Bryden, W. L. (2005). Food Chemistry ,89(2), 309–314. https://doi.org/10.1016/j.foodchem.2004.05.035
Sarwar, G., Botting, H. G. (1993). J. Chromatography. 544. 9–23.
Schwarz, E. L., Roberts, W. L., Pasquali, M. (2004). Clinica Chimica Acta, 343(1–2), 204–209. https://doi.org/10.1016/j.cccn.2004.01.015
Teik, Ng. L., Wong, Y. D., Francis, T., Andersen, G. H. (1991). Ion-exchange chromatography for physiological fluid amino acid analysis. Nutr. Biochem. 2(12), 671–697. https://doi.org/10.1016/0955-2863(91)90066-E
Walker, J. M. (2002). The protein protocols Handbook. Humana Press, 1–1146 https://doi.org/10.1385/1592591698
Williams, A. P., Cockburn, J. E. (1988). Cromatography and Analysis, 1. 9–11.
Downloads
Published
Issue
Section
License
Copyright (c) 2005 Csapó János, Lóki Katalin, Csapóné Kiss Zsuzsanna, Albert Csilla
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
