Degradation Analysis of Human Hemoglobin by High-Performance Liquid Chromatography

Kyung Ho Row; Seung Ki Lee; Y. Polyakova

Journal of Industrial and Engineering Chemistry, Vol.10, No.3, 478-483, May, 2004

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Degradation Analysis of Human Hemoglobin by High-Performance Liquid Chromatography

Kyung Ho Row^†, Seung Ki Lee, and Y. Polyakova

Center for Advanced Bioseparation Technology and Department of Chemical Engineering, Inha University, Incheon 402-751, Korea

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The separation of peptides from human blood and the degradation of hemoglobin was investigated by high-performance liquid chromatography (HPLC). From the experimental results, we determined the optimum mobile phase condition and gradient mode for the separation of peptides from human blood and identified five peptides. Four of the identified peptides were biologically active. The results show that some peptides form and accumulate during the storage of human blood. The kinetics of peptide formation was modeled to that of a first-order reaction. The probability of the short peptides being formed inside human erythrocytes was evaluated by analyzing the peptides using a non-cell model systems (in vitro).

Keywords:degradation;peptides;hemoglobin;human blood;HPLC

[References]

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Ivanov VT, Karelin AA, Philippova MM, Nazimov IV, Pletnev VZ, Biopolym. Peptide Sci., 43, 171 (1997)
Huhmer AFR, Aced GL, Perkins MD, Gursoy RN, Seetharama DS, Larive C, Siahaan TJ, Schoneich C, Anal. Chem., 69, 29R (1997)
Seiler P, J. Chromatogr. A, 852, 273 (1999)
Blishenko EY, Mernenko OA, Yatskin ON, Ziganshin RH, Philippova MM, Karelin AA, Ivanov VT, Biochem. Biophys. Res. Commun., 224, 721 (1996)
Mant CT, Hodges RS, CRC, 938 (1991)
Row KH, Polyakova Y, Lee SK, J. Ind. Eng. Chem., 9(5), 473 (2003)
Vaskovsky BV, Kishenevsky RN, Mikhaleva II, Ivanov VT, Khavinson VK, Morozov VG, Mikhaltsov AN, Anisimov VN, Chemistry of Peptides and Proteins, D. Brandenburg, V. Ivanov, and W. Voelter Eds., Aachen: Verlag Mainz, p. 309 (1993)
Apffel A, J. Chromatogr. A, 712, 177 (1995)
Mints L, Angeletti RH, Nieves E, J. Biomol. Tech., 8, 22 (1997)
Wickner S, Science, 286, 1888 (1999)
Braunitzer G, Gehring-Muller R, Hilschmann N, Hilse K, Hobom G, Rudloff V, Wittmann-Liebold B, Hoppe-Seyler's Z. Physiol. Chem., 325, 283 (1961)
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Abbas A, Sultan C, Zaid ZH, J. Protein Chem., 8.5, 00629-46 (1989)
Abbas A, Zaid ZH, J. Protein Chem., 8.5, 00647-52 (1989)
Lee SK, Polyakova Y, Row KH, Korean J. Biotechnol. Bioeng., 17, 1 (2002)
Dawson RMC, Elliott DC, Elliot WH, Jones KM, Data for Biochemical Research, Clarendon Press, Oxford (1986)
Karelin AA, Filippova MM, Yatskin ON, Blishchenko EY, Nazimov IV, Ivanov V, Russian J. Bioorganic Chem., 24, 241 (1998)
Laidler KJ, Chemical Kinetics, Harper & Row, New York, 3rd Ed. (1987)
House JE, Principles of Chemical Kinetics, Wm C. Brown, Dubuque, IA (1997)
Polyakova Y, Bulanova AV, Vartapetyan RS, Russian Chem. Bull. Int. Ed., 50, 820 (2001)

[Related Articles]

[1] Karelin AA, Philippova MM, Karelina EV, Srtizhkov BN, Grishina GA, Nazimov IV, Ivanov VT, J. Bioorg. Chem., 18, 1 (1993)

[2] Ivanov VT, Karelin AA, Philippova MM, Nazimov IV, Pletnev VZ, Biopolym. Peptide Sci., 43, 171 (1997)

[3] Huhmer AFR, Aced GL, Perkins MD, Gursoy RN, Seetharama DS, Larive C, Siahaan TJ, Schoneich C, Anal. Chem., 69, 29R (1997)

[4] Seiler P, J. Chromatogr. A, 852, 273 (1999)

[5] Blishenko EY, Mernenko OA, Yatskin ON, Ziganshin RH, Philippova MM, Karelin AA, Ivanov VT, Biochem. Biophys. Res. Commun., 224, 721 (1996)

[6] Mant CT, Hodges RS, CRC, 938 (1991)

[7] Row KH, Polyakova Y, Lee SK, J. Ind. Eng. Chem., 9(5), 473 (2003)

[8] Vaskovsky BV, Kishenevsky RN, Mikhaleva II, Ivanov VT, Khavinson VK, Morozov VG, Mikhaltsov AN, Anisimov VN, Chemistry of Peptides and Proteins, D. Brandenburg, V. Ivanov, and W. Voelter Eds., Aachen: Verlag Mainz, p. 309 (1993)

[9] Apffel A, J. Chromatogr. A, 712, 177 (1995)

[10] Mints L, Angeletti RH, Nieves E, J. Biomol. Tech., 8, 22 (1997)

[11] Wickner S, Science, 286, 1888 (1999)

[12] Braunitzer G, Gehring-Muller R, Hilschmann N, Hilse K, Hobom G, Rudloff V, Wittmann-Liebold B, Hoppe-Seyler's Z. Physiol. Chem., 325, 283 (1961)

[13] Perutz MF, Molecular Anatomy, Physiology, and Pathology of Hemoglobin, Molecular Basis of Blood Diseases, C. Stammatagayanopoulus et al. Ed. Philadelphia, Saunders (1987)

[14] Abbas A, Sultan C, Zaid ZH, J. Protein Chem., 8.5, 00629-46 (1989)

[15] Abbas A, Zaid ZH, J. Protein Chem., 8.5, 00647-52 (1989)

[16] Lee SK, Polyakova Y, Row KH, Korean J. Biotechnol. Bioeng., 17, 1 (2002)

[17] Dawson RMC, Elliott DC, Elliot WH, Jones KM, Data for Biochemical Research, Clarendon Press, Oxford (1986)

[18] Karelin AA, Filippova MM, Yatskin ON, Blishchenko EY, Nazimov IV, Ivanov V, Russian J. Bioorganic Chem., 24, 241 (1998)

[19] Laidler KJ, Chemical Kinetics, Harper & Row, New York, 3rd Ed. (1987)

[20] House JE, Principles of Chemical Kinetics, Wm C. Brown, Dubuque, IA (1997)

[21] Polyakova Y, Bulanova AV, Vartapetyan RS, Russian Chem. Bull. Int. Ed., 50, 820 (2001)