ISOLATION AND CHARACTERIZATION OF 2,5-DIPROPIONIC ACID PYRAZINE FROM ALKALINE MODIFIED DELTA-AMINO LEVULINIC ACID AND IDENTIFICATION OF ITS POSSIBLE DERIVATIVES IN HUMAN URINE AND RAT TISSUE HOMOGENATES

When delta aminolevulinic acid is placed in an alkaline solution at a pH of 10 or above, a new compound is formed. It is the purpose of the experiments reported in this research to isolate and characterize this newly formed materials; to determine if such compound or its derivatives can be isolated from urine of patients with Acute Intermittent Porphyria (AIP) and homogenates from liver and bone marrow of rats.; Using an array of physicochemical techniques such as Ultra Violet Spectroscopy, Infrared Spectroscopy, Nuclear Magnetic Resonance Spectroscopy, Mass Spectroscopy and Paper Chromatography the isolated compound was identified as 2,5-Dipropionic Acid Pyrazine.; Possible production of derivatives of 2,5-dipropionic acid pyrazine was investigated in human urine. For this purpose, five individuals were selected from three generations of a family with a history of AIP. Four non related individuals with no history of AIP were used as control.; Partial purification of the urine was achieved using Sephadex G-10, Sephadex G-25, Carboxy Methyl Cellulose and Paper Chromatography. Of all the urine samples examined only that from an asymptomatic AIP patient revealed fractions with similar characteristics to those of 2,5-dipropionic acid pyrazine. Mass spectra of this fraction revealed a parent peak of m/e 195. A structure, 2 ethoxy 5 propionamide pyrazine is tentatively assigned based on the fragmentation pattern.; The mechanism by which 2 ethoxy 5 propionamide pyrazine is formed is unknown. However, this compound may be important in abnormal metabolism of porphyrins associated with inborn enzymatic defects.; An additional investigation was performed with 5-('14)C-delta-ALA using rat liver and bone marrow homogenates. Preliminary results revealed some recovered radioactivity in fractions with similar gel filtration rate as 2,5-dipropionic acid pyrazine.