Measurement of urinary total 13C and 13C urea by isotope ratio mass spectrometry after administration of lactose [13C]-ureide.

A method of measuring total 13C excreted in urine after oral administration of lactose [13C]-ureide was developed using isotope ratio mass spectrometry. Furthermore, a method to measure 13C urea excreted in the urine was developed. Each urine sample collected over a 24 hour period, after administration of the tracer dose, was analysed for both total 13C and 13C urea. Combustion of the dried urine samples allowed measurement of the total 13C content. Treatment of urine samples with urease (EC 3.5.1.5) and analysis by isotope ratio mass spectrometry of the CO2 evolved allowed measurement of 13C urea in the urine sample. The total 13C and 13C urea content of each urine sample, obtained throughout the protocol, were compared to total 13C and 13C urea contents of a urine sample taken before the test. This allowed calculation of the fraction of tracer incorporated into urea and the fraction of tracer excreted in total. Analyses showed that approximately 15% of the dose administered, in terms of 13C, was recovered in the urine over the sampling period. Further analysis for urinary 13C urea showed that less than 1% of the label was incorporated into urea excreted over the sampling period.