Presence of Anti-Pernicious Anemia Principle in Normal Human Urine. ∗

Abstract

In a previous communication 1 we reported a significant modification of the pigeon method of Vaughan, et al., 2 for the laboratory assay of anti-pernicious anemia preparations and showed that normal human urine contains a principle which significantly increases the reticular material in the red blood cells of the pigeon in a manner analogous to the anti-pernicious anemia principle present in liver. At the same time, we reported the preparation of a urinary extract which had a similar effect. In order to test for the identity of the urinary and liver principles, we indicated our plan to study the effect of the urinary extract in pernicious anemia. Due to the well-known present difficulty in securing pernicious anemia patients suitable for assay work, this preliminary report deals with the results obtained with the extract in one patient.

The extract was prepared by concentrating urine to a small volume in vacuo at a temperature of 37°C., adding sufficient 95% ethanol to give a concentration of 70%, concentrating the resulting filtrate in vacuo to a small volume at 37°C, adding sufficient 100% ethanol to give a concentration of 95%, collecting and drying the precipitate, and finally dissolving it in physiological salt solution containing 0.5% phenol. Enough saline solution was used to give a 1:20 ratio for the volumes of the extract and the original urine respectively, although a smaller amount of the solvent would have sufficed. This method is basically that commonly employed in the preparation of parenteral liver extracts.

The volume of this urinary extract necessary to increase significantly the reticular material of the pigeon is approximately equal to that for one commercial parenteral liver extract (3 cc. equivalent to 100 gm. of liver), slightly less than that for another (2 cc. equivalent to 100 gm. of liver), and definitely less than that for a third (20 cc. equivalent to 100 gm. of liver).