Abstract

The importance of deuterium in isotope dilution assay methods as well as in chemical, biochemical, and process development tracing studies has been well established. During the past several years, assay procedures have been developed in this laboratory for such compounds as benzene hexachloride, nicotinic acid, and benzylpenicillin by utilization of the isotope dilution principle. Infrared spectrophotometry has played an important role in these investigations, for through its use was determined the composition of the tracer-assayate mixture, the essential quantity in such assays. (The assayate is defined as the substance for which assay is sought). The introduction of deuterium into the gamma isomer of benzene hexachloride caused a significant change in the infrared spectrum, making possible a direct determination of the composition of the isotopic mixture. Such was also the case with nicotinic acid, but, because of its unfavorable solubility characteristics, the infrared measurement was carried out on the ester, methyl nicotinate. The infrared spectrum of deutero benzylpenicillinic acid, on the other hand, was not sufficiently different from the natural product to form the basis of a precise determination. This difficulty was overcome by subjecting the isotopically mixed benzylpenicillinic acids to an alkaline hydrolysis, isolating qualitatively (but in pure form) the phenylacetic acid, and determining by infrared spectrophotometry the ratio of the two isotopic analogs of phenylacetic acid.

Spectroscopic Analysis of Deuterium in Hydrogen-Deuterium Mixtures

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