A sensitive method for determining nitrogen-15 isotope in urea

Abstract

A method is presented in which the ¹⁵N at.% of urea is determined with high precision on liquid samples containing as little as 10 nmol of urea. The method involves removal of interference from NH₄ ⁺ initially present in the sample by cation exchange. Urea in the sample is subsequently hydrolyzed to NH₄ ⁺ by the enzyme urease. Liberated NH₄ ⁺ is separated from the alkaline sample by diffusion as NH₃ through a helium gas phase where it is finally oxidized to N₂ by reaction with hypobromite iodine. The isotopically labeled N₂ thus formed is mixed with the N₂ initially present in the sample, and the ¹⁵N at.% of urea is calculated from the relative amounts of ¹⁴N¹⁵N and ¹⁵N¹⁵N by means of the isotope pairing principle. The presented method for ¹⁵N-urea analysis proved to be precise (SE < 0.4 at.%, n = 5), when applied on both marine and freshwater samples with an ¹⁵N enrichment >1 at.%, and interference was found only from volatile methyl amines.