Method development for gypenosides fingerprint by high performance liquid chromatography with diode-array detection and the addition of internal standard.
In this paper, a new method for liquid chromatographic fingerprint of saponins in Gynostemma pentaphyllum (THUNB.) MAKINO was developed. The G. pentaphyllum powder was defatted by Soxhlet extraction with petroleum ether and then gypenosides were extracted from the residue with methanol by sonicating. Column chromatography with macro pore resin was then used to separate and enrich gypenosides. HPLC fingerprint analysis of gypenosides fraction was performed on a C18 column, with an isocratic elution of 34% acetonitrile for 60 min at 0.8 ml/min, sample injection volume was 20 microl and the wavelength was 203 nm. To cover the lack of standard compounds, the addition of an internal standard of ginsenoside Rb2 was employed in the gypenosides fingerprint profile. The relative retention time (RRT) and relative peak area (RPA) of the gypenosides peaks in the fingerprint were calculated by setting the ginsenoside Rb2 as the marker compound. The relative standard deviation (RSDs) of RRT of five common peaks vs. ginsenoside Rb2 in precision, repeatability and stability test were less than 1%, and the RSDs of RPA were less than 5%. The method validation data proved that the proposed method for the fingerprint with internal standard of G. pentaphyllum saponins is adequate, valid and applicable. Finally, three batches of crude drug samples collected from Shanxi province were tested by following the established method.
Liu
Fang
F
School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China.
Ren
Dequan
D
Guo
De-an
DA
Pan
Yifeng
Y
Zhang
Huzhe
H
Hu
Ping
P
D002681:China; D002851:Chromatography, High Pressure Liquid; D036145:Ginsenosides / Q000737:chemistry; D029774:Gynostemma / Q000737:chemistry*; D007202:Indicators and Reagents; D000432:Methanol; D010936:Plant Extracts / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D012503:Saponins / Q000737:chemistry; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D036145:Ginsenosides; D007202:Indicators and Reagents; D010936:Plant Extracts; D012503:Saponins; D012997:Solvents; C474918:gypenoside; C048026:ginsenoside Rb2; D000432:Methanol