Case Study #4

LC Method Development for Purine Metabolites in Urine

The Problem:
This group was studying purine metabolism as it related to the formation of kidney stones. A reliable analytical method was needed to determine these metabolites in urine - creatinine, xanthine, allantoin, and uric acid. The presence, absence, and/or elevation of these metabolites can indicate the disease state.

The Solution:
ACCTA, Inc. worked with this group to develop a reliable method for determination of these four metabolites in urine. We also trained the operators on the method, the instrument, and the software.

The Results:
An efficient method was developed to provide the necessary sensitivity and selectivity. As shown in the chromatogram below, analysis was complete in under 12 minutes. We were able to easily separate the compound of interest from the many other matrix components, and optimized the sample preparation prcedures for this column type.

These compounds are difficult to separate by reversed phase LC because they are water soluble and show little retention. In this method, we were able to use an aqueous normal phase (ANP) sytem (also known as HILIC). Gradient programming and wavelength switching were also necessary, but the results allowed easy determination in urine using a relatively simple mobile phase system.

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