Lysimachia capillipes Hemsl (Primulaceae), a folk medicinal plant in China, showed significant anti-tumor activities in vivo and in vitro. Capilliposide B (LC-B) and capilliposide C (LC-C) are the main bioactive components in this plant. To explore their tissue distribution, a reliable bioanalytical method for the quantification of LC-B, LC-C and their bioactive metabolite, capilliposide A (LC-A), in mouse tissues was developed and validated. In this study, the tissue distribution profiles of the three compounds were examined after intravenous administration of pure LC-B and oral administration of total saponins of L. capillipes Hemsl extract (LCE) for 10days. Method validation was conducted over the curve range 10.0-5000ng/mL for all three analytes in various tissue homogenates. The relative standard deviation of intra-day and inter-day precision of the QC samples was <14.7%, and the accuracy ranged from 85.9 to 114.0%. The results indicated that LC-B was rapidly and widely distributed throughout the whole body except for muscle following intravenous administration of LC-B. In addition, LC-A was only in liver, intestine, lung and stomach. After oral administration of LCE, LC-B and LC-C were distributed into various tissues. The highest levels were observed in stomach and intestine.