THE ISOLATION OF MORPHINE ETHEREAL SULFATE FROM URINE OF THE CHICKEN AND CAT

¹ Department of Pharmacology, School of Medicine, Tulane University, New Orleans, Louisiana

Hens were given i.m. doses of morphine sulfate. The urine was collected and filtered through diatomaceous earth. A 100-ml sample was applied to an Amberlite XAD-2 resin column and washed through with 100 ml of water. Morphine and its metabolites were eluted off the resin with methanol. The material which crystallized was collected by filtration. Several such runs of urine samples yielded 96 mg of crystals. Urine from a cat given multiple doses of morphine was collected, pooled and processed by the same method used for the samples from the chicken. Yield was 90 mg. The infrared spectra of the material from the cat and chicken were identical to a sample of morphine ethereal sulfate synthesized chemically by a modification of the method of Stolnikow. The elemental analyses were satisfactory for C, H, N and S. The ultraviolet spectra did not show pH-dependent shifts; these data are consistent with sulfuric acid conjugation of morphine at the 3 phenolic OH position. The procedure for the chicken was quantitated by isolating the radioactive metabolite, and the results indicated that morphine ethereal sulfate was a major metabolite of morphine in the urine of the chicken. Another metabolite appeared to be morphine-3-monoglucuronide. Morphine ethereal sulfate appeared to be a major metabolite in the cat. The isolation procedure applied to rabbit urine yielded crystalline morphine-3-monoglucuronide. The utilitarian value of Amberlite XAD-2 resin for isolating metabolites was demonstrated by the results.