Specific method for determining uric acid in serum using high-performance liquid chromatography and gas...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Clinical Chemistry

Background:

• Accurate quantification of uric acid in serum is crucial for diagnosing conditions like gout and kidney disease.
• Existing methods may lack specificity or require complex sample preparation.

Purpose of the Study:

• To develop and validate a specific and reliable method for serum uric acid quantitation.
• To utilize stable-isotope dilution-mass spectrometry coupled with chromatography for enhanced accuracy.

Main Methods:

• Serum samples were spiked with a known amount of labeled uric acid ([1,3,9-15N]uric acid) as an internal standard.
• Uric acid was purified using anion-exchange chromatography and converted to its tetraethyl derivative.
• High-performance liquid chromatography (HPLC) separated uric acid isomers for mass spectrometry analysis.

Main Results:

• The method achieved specific quantitation of uric acid by measuring the isotope ratio of tetraethyluric acid derivatives.
• Effective separation of isomers was demonstrated using both adsorption and reversed-phase HPLC.
• The technique provides accurate determination of endogenous uric acid concentrations in serum specimens.

Conclusions:

• The described HPLC and stable-isotope dilution-mass spectrometry method offers a robust approach for precise serum uric acid measurement.
• This method can be valuable for clinical laboratories requiring accurate diagnostic information.
• The use of labeled internal standards and specific chromatographic separation enhances analytical reliability.