Isolation and partial characterization of pyridoxal 5'-phosphate hemoglobins by high-performance liquid chromatography as a quality-control method for hemoglobin-based blood substitutes

Journal of Chromatography
M J McGarrityJ C Hsia

Abstract

Pyridoxal 5'-phosphate hemoglobin (PLP-Hb), prepared from hemoglobin and a four-fold excess of pyridoxal 5'-phosphate by the method of De Venuto and Zegna [J. Surg. Res., 34 (1983) 205], has been chromatographically resolved into six components via a quaternary ammonium monobead support. On an analytical scale, the separations have been found to be rapid (ca. 50 min) and highly reproducible. The results also indicate that the preparation of PLP-Hb yields a reproducible product ratio. The potential of the analytical method for the routine quality control of blood substitutes derived from PLP-Hb is discussed. All five of the PLP derivatives (components II-VI), isolated and purified via a combination of conventional and preparative monobead anion-exchange chromatography, gave single peaks when analyzed by high-performance liquid chromatography. Total phosphate analyses indicated that components II and III each contain two PLPs per Hb, IV and V four and VI six.

References

Nov 1, 1975·Analytical Biochemistry·O W van Assendelft, W G Zijlstra
Oct 5, 1977·Journal of Molecular Biology·A ArnoneR Benesch
Jun 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·S C TamJ T Wong
Jan 10, 1986·Journal of Chromatography·J C Hsia, S S Er
Jan 18, 1985·Journal of Chromatography·W F Moo-Penn, D L Jue
Aug 3, 1984·Journal of Chromatography·D RobinetA Funes
Mar 1, 1983·The Journal of Surgical Research·F DeVenuto, A Zegna
Jun 15, 1983·Biochemical and Biophysical Research Communications·K IwasakiY Iwashita
Dec 18, 1981·Journal of Chromatography·T I PristoupilS Ulrych
Feb 1, 1981·The Journal of Surgical Research·L R SehgalG S Moss

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Citations

Jan 1, 1990·Biomaterials, Artificial Cells, and Artificial Organs·P MenuC Vigneron
Mar 20, 1987·Journal of Chromatography·M J McGarrityJ C Hsia

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