A model of thermally generated pH gradients in tapered capillaries

Abstract

Potential applied at the two ends of a tapered capillary can produce an electric field and a pH gradient at the same time. These fields will effect electrophoretic separation, if the diameters of capillaries are not uniform. In some cases these gradients can be used to our advantage. For example, this approach provides a simple way to perform isoelectric focusing without carrier ampholytes as Joule heat produced in the tapered capillary generates a temperature gradient, which in turn produces a pH gradient. A mathematical model is developed to analyze the temperature distribution along the tapered capillary axis. According to the model, the span of the temperature gradient increases with the increase in heat generated per volume of the capillary channel, and the slope of the gradient is determined by the steepness of the taper. The temperature profile predicted by the model is verified by measuring temperatures at the outer wall of the capillary. The calculated pI value of a sample protein, hemoglobin, which focuses in the thermally generated pH gradient, is near to its literature value. The model provides us with a better understanding of the heat transfer in a tapered capillary and theoretically demonstrates the potential of capillary isoelectric focusing separation with a tapered channel.

References

1. A. Kolin, in Isoelectric Focusing, ed. N. Catsimpoolas, Academic Press, New York, USA, 1976, pp. 1–11.
2. S. Hjerten and M-D. Zhu, J. Chromatogr., 1985, 346, 265.
3. S. Hjerten, in Capillary Electrophoresis: Theory and Practice, ed. P. D. Grossman and J. C. Colburn, Academic Press, New York, USA, 1992, pp. 191–214.
4. T. Pritchett, in Molecular Biology: Current Innovations and Future Trends, ed. A. M. Griffin and H. G. Griffin, Horizon Scientific Press, Norwich, Norfolk, UK, 1995, pp. 127–145.
5. X. Liu, Z. Sosic and I. S. Krull, J. Chromatogr., 1996, 735, 165.
6. H. Rilbe, J. Chromatogr., 1978, 159, 193.
7. R. A. Mosher, W. Thormann, A. Graham and M. Bier, Electrophoresis, 1985, 6, 545.
8. O. Sova, J. Chromatogr., 1985, 320, 15.
9. R. L. Prestidge and M. T. Hearn, Anal. Biochem., 1977, 97, 95.
10. A. Chrambach and N. Y. Nguyen, in Electrofocusing and Isotachophoresis, ed. B. J. Radola and D. Graesslin, Walter de Gruyter & Co., Berlin, Germany 1977, pp. 51–58.
11. L. M. Hjelmeland and A. Chrambach, Electrophoresis, 1983, 4, 20.
12. G. V. Troitski, V. P. Savialov, I. F. Kirjukhin, V. M. Abramov and G. J. Agitski, Biochim. Biophys. Acta, 1975, 400, 24.
13. S. J. Luner and A. Kolin, Proc. Natl. Acad. Sci. U.S.A., 1970, 66, 898–903.
14. C. H. Lochmuller, S. J. Breiner and C. S. Ronsick, J. Chromatogr., 1989, 480, 293.
15. P. Lundahl and S. Hjerten, Ann. N. Y. Acad Sci., 1973, 209, 94.
16. C. H. Lochmuller and C. S. Ronsick, J. Chromatogr., 1991, 249, 297.
17. J. Pospichal, M. Deml and P. Bocek, J. Chromatogr., 1993, 638, 179.
18. J. Pawliszyn and J. Wu, J. Microcolumn Separations, 1993, 5, 397.
19. E. Grushka, R. M. McCormik and J. J. Kirkland, Anal. Chem., 1989, 6, 241.
20. P. D. Grossman, in Capillary Electrophoresis: Theory and Practice, ed. P. D. Grossman and J. C. Colburn, Academic Press, New York, USA, 1992, pp. 5–9.
21. H. S. Carslaw and J. C. Jaeger, Conduction of Heat in Solid, Clarendon Press, Oxford, UK, 2nd edn., 1986, pp. 133–135.
22. M. Mikheyev, Fundamentals of Heat Transfer, Mir Publishers, Moscow, Russia, 1968, pp. 73–81.
23. A. Palm, C. Lindh, S. Hjerten and J. Pawliszyn, Electrophoresis, 1996, 17, 766.
24. P. G. Righetti, Isoelectric Focusing: Theory, Methodology and Applications, Elsevier Biomedical Press, Amsterdam, The Netherlands, 1983.
25. L. P. Vonguyen, Isoelectric Focusing With Cone-shaped Capillaries, MSc Thesis, University of Waterloo, Waterloo, Canada, 1995.
26. R. G. Bates and H. B. Hetzer, J. Phys. Chem., 1961, 65, 667.
27. D. D. Perrin and B. Dempsey, Buffers for pH and Metal Ion Control, Champman and Hall Ltd., London, UK, 1974, pp. 157–163.
28. R. M. C. Dawson, D. C. Elliott, W. H. Elliott and K. M. Jones, Data for Biochemical Research, Clarendon Press, Oxford, UK, 3rd edn., 1986, pp. 423–425.
29. S. Fredriksson, in Electrofocusing and Isotachophoresis, ed. B. J. Radola and D. Graesslin, Walter de Gruyter & Co., Berlin, Germany, 1977, pp. 71–83.
30. P. G. Righetti, G. Tudor and K. Ek, J. Chromatogr., 1981, 220, 115.