Abstract
We evaluated the biochemical characteristics of endogenous fluorescent substances. Ex 380 nm/Em 440 nm and Ex 400 nm/Em 460 nm, present in sera of patients with chronic renal failure (Clin. Chem. 31:1988, 1985). Sera from 23 patients with chronic renal failure (CRF) and from 10 normal subjects were filtered through ultrafiltration membranes (cutoff limit of 500 Da). Fluorescence intensity of the aforementioned substances was significantly elevated as compared to normals (p < 0.001). Fluorescence characteristics of these substances remained unaltered after ultrafiltration and treatment with beta-glucuronidase. Extraction of these fluorescent compounds with organic solvents (dichloromethane, ethyl acetate, chloroform:methanol) could not be achieved after ultrafiltrates were subjected to 6N hydrochloric acid (HCl) hydrolysis. In addition, treatment with 6N HCl enhanced fluorescence intensity without altering fluorescence excitation/emission maxima. Removal of fluorescence could be accomplished in toto by adsorption onto activated charcoal with subsequent recovery from charcoal by treatment with sodium hydroxide, pH 12 (Ex 380 nm: 51.1%, Ex 400 nm: 91.8%). Analysis of alkali-treated specimens by high performance liquid chromatography demonstrated that peptides associated with these fluorescent substances were denatured, although fluorescence at these previously described excitation/emission maxima persisted. Our studies indicate that the unique fluorescence observed in the sera of patients with CRF is not an intrinsic characteristic of a specific peptide or its amino acids, but rather an inherent property of fluorescent molecules which may bind to these peptides.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
H. Mabuchi, and H. Nakahashi, Liquid-chromatographic profiling of endogenous fluorescent substances in sera and urine of uremic and normal subjects, Clin. Chem. 29:675–677 (1983).
H. A. Schwertner, Isolation and chromatographic analysis of unidenti fied fluorescence in biological fluids of patients with chronic renal disease. Nephron 31:209–211 (1982).
J. S. Swan, E. Y. Kragten, and H. Veening, Liquid-chromatographic study of fluorescent materials in uremic fluids, Clin. Chem. 29:1082–1084 (1983).
M. Shaykh, N. Bazilinski, D. S. McCaul, S. Ahmed, A. Dubin, T. Musiala, and G. Dunea, Fluorescent substances in uremic and normal serum, Clin. Chem. 31:1988–1992 (1985).
N. Bazilinski, M. Shaykh, S. Ahmed, T. Musiala, R. H. Williams, A. Poulos, A. Dubin, and G. Dunea, Amino acid composition of uremic middle and low molecular weight retention products, Proc. Symposium Uremic Toxins (1986).
J. P. Monti, P. Gallice, A. Crevat, and A. Murisasco, Identification by nuclear magnetic resonance and mass spectrometry of a glucuronic acid conjugate of o-hydroxybenzoic acid in normal urine and uremic plasma, Clin. Chem. 31:1640–1643 (1985).
O. S. Wolfbeis and M. Leiner, Mapping of the total fluorescence of human blood serum as a new method for its characterization. Anal. Chim. Acta 167:203–215 (1985).
H. A. Schwertner and S. B. Hawthorne, Albumin-bound fluorescence in the serum of patients with chronic renal failure, Clin. Chem. 26:649–652 (1980).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1987 Plenum Press, New York
About this chapter
Cite this chapter
Williams, R.H. et al. (1987). Biochemical Elucidation and HPLC Fractionation of Fluorescent Peptides in Patients with Chronic Renal Failure. In: Ringoir, S., Vanholder, R., Massry, S.G. (eds) Uremic Toxins. Advances in Experimental Medicine and Biology, vol 223. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5445-1_32
Download citation
DOI: https://doi.org/10.1007/978-1-4684-5445-1_32
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-5447-5
Online ISBN: 978-1-4684-5445-1
eBook Packages: Springer Book Archive