Objective To establish a quantitative analysis of multi-components by single-marker (QAMS) for determination of four components (adenosine, uridine, guanosine, (R, S)-goitrin) in Isatidis Radix granules.Method The QAMS for Isatidis Radix granules was established and validated and adenosine was selected as the internal reference substance.The relative correction factor (RCF) of other three components were calculated.The contents of four components were determined by both external standard method and QAMS.The QAMS method was validated through comparison of the results obtained by the two different methods.Results The RCFs of uridine, guanosine and (R, S)-goitrin all showed good reproducibility within certain ranges.For Isatidis Radix granules, uridine, guanosine and (R, S)-goitrin, there was no significant difference between the quantitative results of the two methods.Conclusions The QAMS method is simple, feasible and credible to be used for comprehensive quality control of Isatidis Radix granules.