This study was undertaken to determine the astragaloside content in Qishen Huoxue Granule (QHG) by HPLC and to investigate the protective effect of astragaloside in septic rats. Methods: Dikma Diamonsil (TM) C-18 column was used. Mobile phase was methanol: acetonitrile (42: 58) and detection was performed at 205 nm. Healthy rats were randomly divided into three groups, group A: sham operation (SO) group; group B: cecal ligation and puncture (CLP) group; and group C: CLP QHG treatment group. 72 h after model establishment, plasma tumor necrosis factor-alpha (TNF-alpha), plasma and peritoneal albumin concentrations, as well as mesenteric venular flow velocity in each group were measured and degree of intestinal mucosal injury was observed. Average recovery was 100.7%, with a RSD of 2.20%. Sample concentration exhibited good linearity with peak area within a 0.01 similar to 0.2 mg/mL range. After CLP, plasma TNF-alpha concentration and albumin leakage in rats increased markedly; such increases were more significant in group B than group C (P<0.01). Changes in intestinal mucosal scores also presented the same trend (P<0.01). Mesenteric venular flow slowed down and plasma albumin concentration lowered; such decreases were more significant in group B than group C (P<0.05). QHG can reduce the inflammatory response and intestinal mucosal injury in rats with CLP-induced sepsis.