Abstract
Investigations concerning the blood AMPS pattern have been relatively few, probably owing to low AMPS concentration.
An average AMPS uronic acid level of 206 μg% in normal human plasma has been reported(1), whereas an average concentration of 277 μg% was observed in normal serum (2). On paper chromatography of an AMPS fraction isolated from serum, 2 components, both of which were labile to testicular hyalu-ronidase, were observed(2). The chromato-graphic characteristics of one component resembled that of chondroitin sulfate (ChS). Schiller(3) presented conclusive evidence for ChS-A being the main AMPS of normal human plasma. Starting from several liters of human plasma she obtained 2 AMPS fractions. The nature of the electrophoretically slower fraction remained unresolved. Apparently it was heterogeneous and it probably contained hyaluronic acid (HA) as one component. In the serum of 2 patients with reticulum cell sarcoma and neuroblastoma, respectively, the presence of HA has been demonstrated (4).
Zone electrophoresis on cellulose acetate of AMPS mixtures has proved to be a sensitive analytical tool providing good separations (5,6). To the best of the authors' knowledge this method has not been applied to the analysis of blood AMPS.
Materials and methods. In an investigation undertaken to study the AMPS pattern of normal human serum and serum in various disease states, the AMPS fraction from 10-15 ml of serum obtained from healthy blood donors was isolated by a modification of the procedure of Bollet et al(2). The final precipitation step with protamine was replaced by an alcohol precipitation using 4.5 vol. of 96% ethanol saturated with sodium chloride in the presence of 0.5 N acetic acid. The precipitate was taken up in 2 ml of 0.05 N NaOH, 0.5 ml of which was dialyzed for about 72 hours against running tap water and then evaporated to dryness under reduced pressure.
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