High-performance liquid chromatography and liquid
chromatography-mass spectrometry analysis
The ginsenoside composition of the GS sample was analyzed by
high-performance liquid chromatography (HPLC; Agilent Technologies,
Billerica, MA, USA) with
a
diode-array detector (Ultimate3000, DIONEX, Sunnyvale, CA, USA) using
the Elite Kromasil C18 column (4.6×250 mm, 5 µm). The mobile phase
condition was ultrapure water (solvent A) and acetonitrile (solvent B;
Sigma-Aldrich), and the column temperature was set at 35°C. The flow
rate was 1.0 mL/min, and the chromatograms were obtained using a UV/VIS
detector at 203 nm. According to the retention time and chromatographic
peak area of ginsenoside standards, 18 ginsenosides were identified and
quantified as Rg1 (11.23%), Re (18.32%), Rf (4.37%), Rb1 (5.96%), Rc
(10.34%), Rh1 (1.40%), Rb2 (3.15%), Rb3 (0.57%), F1 (0.22%), Rd
(7.42%), F3 (2.83%), Rk3 (6.30%), S-Rg3 (2.46%), R-Rg3 (1.00%), PPT
(0.16%), Rk1 (1.72%), Rg5 (2.31%), and Rh2 (1.68%) in the GS sample(Supplementary Fig. 1A and 1B) . Based on mobile phase and
elution gradient from HPLC, LC-mass spectrometry (LC-MS) was performed
on the Thermo LTQ XL system (Thermo Fisher Scientific, Carlsbad, CA,
USA) with the Agilent ZORBAX SB-C18 column (2.1×100 mm, 1.7 µm) to
obtain the chromatogram of different ginsenosides in the negative ion
mode. MS analysis conditions were set as follows: gas temperature
270 °C,
electric voltage 3.5 kV and scan
range from 165 to 2000 m/z. The MS chromatogram and properties of
different ginsenosides from GS are shown in Supplementary Fig.
2A and 2B .