UPLC-MS/MS Method for Determination of Uric Acid and Creatinine in Serum and Urine of Hyperuricemic Mice

An ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS) method for the determination of uric acid and creatinine in serum and urine" was established and applied to screening of anti-gout medicines. The automatic sampler and the pump of an LC apparatus were used to inject sample and mobile phase into a chromatography guard column which was used to remove most interference components and enrich the target compounds. MS/MS analysis was carried out in positive ion mode and in multiple reaction monitoring(MRM) mode for creatinine, linearity of the MS/MS assay without and with matrix were 0.03-400 mu mol/L and 0.2-400 mu mol/L of creatinine, with a limit of detection (LOD) of 0.03 and 0.2 mu mol/L, respectively. The uric acid was analyzed in negative ion mode, linearity of the MS/MS assay without and with matrix were 0.1-350 mu mol/L and 0.5-300 mu mol/L of uric acid, with a LOD of 0.1 and 0.5 mu mol/L, respectively. An analytical recovery of this method was 91.8%-103.7% and the RSDs for intra-day and inter-day were less than 5.9% and 6.8% respectively. The results meet the requirements for a bioanalytical assay. Subsequently, the established method was applied to screening of anti-gout medicines. The results showed that ermiaowan, Phellodendri Chinensis Cortex and Atractylodis Rhizoma were able to reduce blood uric acid levels at some degree compared with the positive control drugs, including allopurinol and benzbromarone. In addition, they can significantly reverse the renal damage and protect the renal function.