GSH and Cys metabolites of microcystin-RR（MC-RR） were synthesized chemically by the monitoring of high liquid chromatography-electrospray ionization ion trap mass spectrometry（HPLC-ESI-ITMS）.Based on the enrichment and purification of AccuBOND C18 solid phase extraction（SPE）,the purity of these s-conjugates were higher than 95% by HPLC.In this study,the optimum reaction time and SPE cartridges treatment procedure were also studied,and 60 min was obtained as the optimal time for the s-conjugates synthesis.Extraction parameters including proportions of methanol in elution and washing solution were optimized,and the best mean recoveries were obtained by 90% methanol in elution after water washing,and their recoveries were in the ranges of 86.4%±1.5% and 90.2%±2.8% for GSH conjugates of MC-RR（MC-RR-GSH） and Cys conjugates of MC-RR（MC-RR-Cys）,respectively.These metabolites were subsequently confirmed by high phase liquid chromatography-electrospray ionization ion trap mass spectrometry（HPLC-ESI-ITMS）.The fragment ions of the metabolites of MC-RR were characterized,and the most aboundant ions for MC-RR-GSH and MC-RR-Cys were at m/z 673.6 and m/z 580.5,respectively,and these ions were further selected as precursor ion in MS/MS,while the product ions of these parent ions were at m/z 609.0/536.9 and 519.4/520.8,respectively.These purified metabolites were successfully applied as standard compounds in the quantification of MC-RR-GSH,MC-RR-Cys and MC-RR in the kidney of commom carp,and the contents(dry weight) were obtained at the levels of undetected（ND）,1.587 5,0.001 5 μg?kg-1,respectively.Futhermore,these metabolites could be used as significant reference substances in various studies,and the cleavage rules of MC-RR-GSH and MC-RR-Cys could provide some metabolic mechanism of microcystin-RR.