ABSTRACT
OBJECTIVE: To develop an intergrated urinary metabonomic strategy based on RP-UPLC-MS and HILIC-UPLC-MS method and to investigated the mechanism of osteoporosis and the prevention effect of glucocorticoid osteoporosis of Gushudan. METHODS: The RP-UPLC-MS and HILIC-UPLC-MS method were developed for rat urine metabolite profiling study among control group, model group and Gushudan treatment group. Principal component analysis (PCA) were used to find the biomarkers, further more, to explore the mechanism of the prevention effect on glucocorticoid osteoporosis of Gushudan. RESULTS: In the mode of RP-UPLC-MS, low polarity metabolites like phenylacetylglycine, N2-succinic acid-L-ornithine were found while relatively high polarity metabolites like betaine, hypoxanthine were found in the mode of HILIC-UPLC-MS. And twenty-two potential biomarkers have been totaly found in the urine of glucocorticoid osteoporosis, primiarily related to amino acid metabolism, energy metabolism, lipid metabolism, intestinal flora metabolism and kidney damage. Gushudan has intervention effects on rats with osteoprosis via the regulation of multiplemetabolic pathways. CONCLUSION: The combination of RP-UPLC-MS and HILIC-UPLC-MS method can give a more comprehensive profiling of endogenous metabolites and it also indicates metabonomic strategy has become a powerful tool to evaluate the overall efficacy of traditional Chinese medicine (TCM) and to calrify the mechanism of TCM.