High Performance Liquid Chromatography Coupled with Post-Column Iodine-Azide Reaction for the Determination of Benzylthiouracil in Urine

Abstract

The reaction between iodine and azide ions induced by benzylthiouracil was applied as a post-column detection system for determination of benzylthiouracil in urine. Neither extraction, nor preconcentration of the sample are necessary. The reproducibility, linearity, and recovery were evaluated under the optimum conditions. The benzylthiouracil standards added to normal urine show that the response of the detector, set at 350 nm (corresponding to unreacted iodine in the post-column iodine-azide reaction), was linear within the concentration range of 0.4-4.5 nmol · mL⁻¹ urine. Recovery and the relative standard deviation values for precision within the calibration range were from 94 to 104% and from 1 to 4.5%, respectively. Lower limits of detection (LLD) and quantitation (LLQ) were 0.3 and 0.4 nmol · mL⁻¹ urine, respectively.

Keywords:

• Benzylthiouracil
• High performance liquid chromatography
• Iodine-azide reaction
• Post-column reaction
• Urine

ACKNOWLEDGMENTS

The presented experiment was conducted with the support of grant No. N204 14732/3728 from the Ministry of Science and High Education during 2007–2009, Poland. The author wishes to thank Prof. W. Ciesielski.