A Novel Quantitative Method for Recovering Precipitated Uric Acid in Urine and Analysis by LC–MS/MS

Abstract

Aim: Precipitation of uric acid (UA) in human urine had caused poor recovery and underestimation of UA in study samples. Results: A UA recovery method was developed for analyzing frozen urine samples that potentially contain precipitated UA. Samples were treated with 3% v/v of a solution containing 8 M potassium hydroxide and 20% v/v of ammonium thioglycolate. The high pH enables rapid dissolution of UA crystals, while the thioglycolate protects UA from oxidation at high pH. The treated samples were diluted 20-fold with 50 mM ammonium chloride and analyzed immediately by LC–MS/MS or stored at -20°C prior to analysis. Conclusion: Complete recovery of UA was achieved. Thioglycolate stabilized UA at pH 13 for at least 4.5 h at room temperature. Both accuracy and precision satisfy US FDA bioanalytical method validation (BMV) guidelines.

Keywords::

• ammonium thioglycolate
• recovery
• stability
• supersaturation
• uric acid
• urine

Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Acknowledgements

The authors thank R Winnike, S Liu and C Lee for insightful discussions.