, New York, NY, USA) or by spraying 10% aqueous H2SO4 on the developed plate followed by heating. Optical rotations were measured using a JASCO P-1010 digital polarimeter (Jasco, Tokyo, Japan). A Shimadzu GCMS-QP2010 Plus (Shimadzu, Tokyo, Japan) mass spectrometer (MS) was used for gas chromatography (GC)/MS experiments.

Fast atom bombardment (FAB)/MS spectrum was recorded on a spectrometer (JMS-700; JEOL, Tokyo, Japan). IR spectra were obtained from a PerkinElmer spectrum one Fourier transform-IR spectrometer (PerkinElmer, Buckinghamshire, Caspase inhibitor UK). NMR spectra were recorded on a Varian Inova AS 400 spectrometer (400 MHz; Varian, Palo Alto, CA, USA). The dried and powdered aerial parts of hydroponic P. ginseng find more (6.27 kg) were extracted with 80%

MeOH (30 L × 3) at room temperature for 24 h. The extracts were filtered through a filter paper and evaporated under reduced pressure at 45°C to yield 1.4 kg of extract. The extract was poured into H2O (3 L) and then extracted with ethyl acetate (EtOAc; 3 L × 3) and n-butanol (n-BuOH; 2.6 L × 3) successively. Each layer was concentrated under reduced pressure to obtain EtOAc (75 g), n-BuOH (470 g), and H2O (855 g) fractions. The EtOAc fraction (75 g) was applied on a silica gel column (φ 14 × 16 cm) and eluted with CHCl3–MeOH (30:1, 60 L) and CHCl3–MeOH–H2O (15:3:1, 136 L) to obtain 24 fractions (HPE1 to HPE24). Fraction HPE9 (9.28 g; Ve/Vt = 0.10–0.16, where Ve refers to the volume of eluent for the corresponding fraction and Vt represents the total elution volume) was applied on a silica gel column (φ 7 × 15 cm) using n-hexane–EtOAc (1:2, 28 L) as eluent to obtain 13 fractions (HPE9-1 to HPE-9-13). Fraction HPE9-10 (4.47 g,

Ve/Vt = 0.24–0.98) was further fractionated on an octadecyl silica gel (octadecylsilane Elongation factor 2 kinase or ODS) column (φ 4.5 × 5 cm, MeOH–H2O = 15:1, 4 L) to produce nine fractions (HPE9-10-1 to HPE9-10-9) including 2(S)-1-O-linoleoyl-2-O-linoleoyl-3-O-β-d-galactopyranosyl-sn-glycerol [4, HPE9-10-4, 141.6 mg; Ve/Vt = 0.24–0.29, TLC Rf = 0.25 (RP-18 F254S, MeOH–H2O = 50:1), Rf = 0.50 (Kieselgel 60 F254, n-hexane–EtOAc = 1:30)]. Fraction HPE9-10-2 (3.14 g, Ve/Vt = 0.06–0.14) was further fractionated on the ODS column (φ 4 × 6 cm, acetone–acetonitrile–H2O = 2:2:1, 3.6 L) to yield 10 fractions (HPE9-10-2-1–HPE9-10-2-10) including (2S)-1-O-linolenoyl-2-O-linolenoyl-3-O-β-d-galactopyranosyl-sn-glycerol [3, HPE9-10-2-9, 446.0 mg, Ve/Vt = 0.38–0.55, TLC Rf = 0.50 (RP-18 F254S, acetone–acetonitrile–H2O = 7:3:1), Rf = 0.55 (Kieselgel 60 F254, CH2Cl2–MeOH = 10:1)]. Fraction HPE15 (5.49 g, Ve/Vt = 0.34–0.36) was further fractionated on the ODS column [φ 4.5 × 12 cm, MeOH–H2O = 3:2 (1.0 L) → 2:1 (2.5 L) → 3:1 (5.2 L) → 5:1 (2.0 L)] to yield 25 fractions (HPE15-1–HPE15-25).