Retention Mechanism of Mononucleotides with Buffer in RP-HPLC

Longmei Jin; Du Young Choi; Yining Wang; Wha Seung Ahn; Dae Ki Choi; Kyung Ho Row

Journal of Industrial and Engineering Chemistry, Vol.11, No.4, 615-623, July, 2005

Export Citation

Retention Mechanism of Mononucleotides with Buffer in RP-HPLC

Longmei Jin, Du Young Choi, Yining Wang, Wha Seung Ahn , Dae Ki Choi¹, and Kyung Ho Row^†

Center for Advanced Bioseparation Technology and Department of Chemical Engineering, Inha University, Incheon 402-751, Korea
¹Clean Technology Research Center, Korea Institute of Science and Technology, Seoul 136-791, Korea

E-mail:

To estimate the retention equilibrium constant of ionized solute by the buffers in the mobile phase, RP-HPLC experiments were performed under isocratic conditions. Four nucleotides (5'-UMP, 5'-GMP, 5'IMP, 5'TMP) were used as solutes. The first absolute moment and second central moment were determined from the elution curves by means of moment analysis. The retention equilibrium constant decreases upon increasing the concentration of the buffers. Three different mass transfer processes (i.e., δax, δf, and δd) that take place in the column were investigated using different buffers. The results indicate that δax and δf had large effects on μ'2, but the contribution of δd to μ'2 was negligible. Inaddition, we found that surface diffusion plays a dominant role in intraparticle diffusion.

Keywords:retention equilibrium;nucleotides;moment analysis

[References]

Miyabe K, Guiochon G, J. Sep. Sci., 26, 155 (2003)
Guiochon G, Golshan-Shirazi S, Katti AM, Fundamentals of Preparative and Nonlinear Chromatography, Academic Press, Boston (1994)
Ruthven DM, Principles of Adsorption & Adsorption Processes, John Wiley and Sons, New York (1984)
Suzuki M, Adsorption Engineering, Kodansha/Elsevier, Tokyo/Amsterdam (1990)
Treybal RE, Mass-Transfer Operaations, McGraw-Hill, New York (1980)
Bird RB, Stewart WE, Lightfoot EN, Transport Phenomena, John Wiley & Sons, New York (2002)
Lee JW, Row KH, HWAHAK KONGHAK, 38(5), 626 (2000)
Miyabe K, Suzuki M, AIChE J., 38, 901 (1992)
Row KH, Lee CH, Kang JH, Biotechnol. Bioprocess Eng., 7, 16 (2002)
Miyabe K, Suzuki M, Ind. Eng. Chem. Res., 33(7), 1792 (1994)
Miyabe K, Suzuki M, AIChE J., 41(3), 548 (1995)
Kanji M, Sayuri S, Georges G, J. Chromatogr. A, 919, 231 (2001)
Choi DY, Row KH, J. Ind. Eng. Chem., 10(6), 1052 (2004)

[Related Articles]

[1] Miyabe K, Guiochon G, J. Sep. Sci., 26, 155 (2003)

[2] Guiochon G, Golshan-Shirazi S, Katti AM, Fundamentals of Preparative and Nonlinear Chromatography, Academic Press, Boston (1994)

[3] Ruthven DM, Principles of Adsorption & Adsorption Processes, John Wiley and Sons, New York (1984)

[4] Suzuki M, Adsorption Engineering, Kodansha/Elsevier, Tokyo/Amsterdam (1990)

[5] Treybal RE, Mass-Transfer Operaations, McGraw-Hill, New York (1980)

[6] Bird RB, Stewart WE, Lightfoot EN, Transport Phenomena, John Wiley & Sons, New York (2002)

[7] Lee JW, Row KH, HWAHAK KONGHAK, 38(5), 626 (2000)

[8] Miyabe K, Suzuki M, AIChE J., 38, 901 (1992)

[9] Row KH, Lee CH, Kang JH, Biotechnol. Bioprocess Eng., 7, 16 (2002)

[10] Miyabe K, Suzuki M, Ind. Eng. Chem. Res., 33(7), 1792 (1994)

[11] Miyabe K, Suzuki M, AIChE J., 41(3), 548 (1995)

[12] Kanji M, Sayuri S, Georges G, J. Chromatogr. A, 919, 231 (2001)

[13] Choi DY, Row KH, J. Ind. Eng. Chem., 10(6), 1052 (2004)