| Background and ObjectiveThe incidence rate of hyperuricemia and gout increased rapidly in recently years. The antihyperuricemia treatment is necessary to reduce the gouty arthritis and nephropathy. But there are few of hypouricemic drugs that are safe, effective and economical in clinic. In China, though there is a long history of gout treatment using traditional Chinese medicine (TCM), no potent hypouricemic TCM has been found. So it is important to look for new hypouricemic drugs, especially hypouricemic TCM.The first objective of our research is to establish a system that applies to screen new hypouricemic drugs. The system includes a reliable hyperuricemia animal model and special RP-HPLC determination method. The second objective of our research is to look for potent hypouricemic TCM, reference to the TCMtheory. The research can be divided into 4 parts.Part 1: Simultaneous determination of purine metabolites and creatinine in blood and urine by reversed-phase high-performance liquid chromatography.Oxonic acid, an uricase inhibitor used in establishing hyperuricemia animal model, can disturb current uric acid biochemical assay. So it is necessary to find a new uric acid assay that will not be disturbed. High performance liquid chromatography (HPLC) is accurate and sensitive, can determine different compounds simultaneously. In part 1, a practical one-step RP-HPLC method thatcan simultaneous determine hypoxanthine, xanthine, uric acid, allantoin and creatinine in blood and urine was established.Part 2: Studies on rat hyperuricemia model.There is no generally accepted hyperuricemia animal model. In most patients, the uric acid excretion is decreased. But in existing models, the uric acid excretion is increased significantly. There is obvious difference between models and patients. In part 2, a new hyperuricemia model that is more similar to patient is established by oxonic acid and pyrazinamide.Part 3: Statistical Analysis of Traditional Chinese Medicine Prescriptions for gout.How to identify the candidate rapidly from thousands of TCM is an important question. The experiences of TCM theory is valuable, but because of obvious different opinion about gout, the learning must be selective. In part3, according to definite standards, 45 TCM prescriptions were selected and several candidates were identified by statistical analysis.Part 4: Studies on the hypouricemic effect of several traditional Chinese medicines on hyperuricemia rat model.On the basis of other three parts, in part 4, several TCMs were administrated to the hyperuricemia rat and the changes on uric acid metabolism were observe.MethodsPart 1: The reversed-phase column was a Fuji Silysia Cig column (250X4.6 mm, made in China). The mobile phase was phosphate buffer (0.05M, pH=6.0). Chromatography was achieved at a flow rate of 0.5 ml/min and injection volume of 20 u 1. The column effluent was monitored from 210nm to 400nm. Total analysis time of 30 minutes. The plasma was deproteinized with 5% perchloric acid and the urine was diluted and filtrated before determination.Part 2: (1). Twenty-four rats were divided into 4 groups and wereadministrated uric acid (UA group), oxonic acid (OA group), oxonic acid + uric acid (OA+UA group) and oxonic acid + pyrazinamide (OA+PZA group) i.p. separately twice daily to establish hyperuricemia model. The duration was 7 days. The uric acid metabolism indexes of baseline and at endpoint were compared. (2) Eighteen rats were divided into 3 groups and were administrated benzbromarone (BEN group), allopurinol (ALL group) or distilled water (control group) separately for 7 days. AH the groups were administrated OA+PZA at the same time. The uric acid metabolism indexes of baseline and at endpoint were compared.Part 3: (1) Search TCM prescriptions from gout treatment clinic researches in medical journals. All the selected research should have clearly diagnosis, scientific research methods, reliable hypouricemic effect and have not disturbing factors. 45 TCM pr...
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