The glycosyl sequence of gypenoside A (A) isolated from Gynostemma pentaphyllum Makino was determinated by TLC-FABMS. The gypenoside A was hydrolysed with cellulase and the hydrolysate showed eight spots on a TLC plate, and the FABMS analysis of these spots indicated they corresponded to gypenoside A, A-Rham, A-Glu, A-Rham-Glu, A-Rham-Glu, A-Glu-Glu, A-Rham-2Glu, A-Rham-2Glu in order. Two chains can be determined to be Rham-Glu, and Glu-Glu, respectively. So the glycosyl sequence of gypenoside A was identical of gypenoside XLIII.