An improved high performance liquid chromatographic method was developed to determinate the active triacylglycerol principles ofPanax pseudo-ginsengWall. var. notoginseng Hoo & Tseng extracts. Chromatographic analysis is achieved on an isocratic system consisting of a polymeric reverse phase C18column with a mobile phase of acetonitrile-methanol (50:50, v/v) to elute the trilinolein and triolein. The eluted triacylglycerol are detected at 205 nm. The linear calibration range was 0.5–20 and 20–500 μg/mL for trilinolein and triolein, respectively. The detection limit was 0.1 μg/mL for trilinolein and 10 μg/mL for triolein. It was found that n-hexane extracts ofPanax pseudo-ginsengWall. var. notoginseng Hoo & Tseng contained 0.21 ± 0.021 mg/g of trilinolein. However, triolein was not detectable in the same herbal extract.