Fig. 1. Chemical structures of target compounds from Rhizoma Anemarrhenae.

Fig. 2. HPLC chromatograms of crude extracts from Rhizoma Anemarrhenae (A) and HSCCC peak fractions (B–E). Column: YWG ODS C₁₈ column (200 × 4.6 mm I.D., 10 μm); mobile phase: methanol-phosphoric acid (0.1%) (Methanol: 0–5 min, 10%; 5–18 min, 10–50%; 18–28 min, 50–100%); flow rate: 1.0 ml min⁻¹; detection wavelength: 254 nm.

Fig. 3. HSCCC chromatograms of crude extracts (A) and refined sample (B) from Rhizoma Anemarrhenae. Conditions of: (A) Two-phase solvent system: n-butanol–acetic acid (1%) (1:1, v/v); mobile phase: the lower phase; flow rate: 2.0 ml min⁻¹; revolution speed: 700 rpm; detection wavelength: 254 nm; sample size: 500 mg of crude extracts dissolved in 5 ml of the lower phase; separation temperature: 25 °C. (I) neomangiferin (collected during 47–61 min); (II) mangiferin (collected during 144–238 min). Conditions of: (B) Two-phase solvent system: light petroleum–ethyl acetate–methanol–water (1:1:1.2:0.8, 1:1:1.4:0.6, v/v) in gradient elution; Stationary phase: upper organic phase of 1:1:1.2:0.8; mobile phase: lower phase of 1:1:1.2:0.8 and 1:1:1.4:0.6 used in gradient elution mode as follows: 0–100 min, only the lower phase of 1:1:1.2:0.8 system; 100–130 min, the volume ratio of 1:1:1.2:0.8 and 1:1:1.4:0.6 system was continuously changed from 100:0 to 0:100; flow rate: 2.0 ml min⁻¹; revolution speed: 850 rpm; detection wavelength: 254 nm; sample size: 80 mg of refined sample dissolved in 3 ml of the upper phase of light petroleum–ethyl acetate–methanol–water (1:1:1.2:0.8, v/v); separation temperature: 25 °C. (I) cis-hinkiresinol (collected during 54–63.5 min); (II) (-)-4′-O-methylnyasol (collected during 162–176 min).

The K-values of target components measured in several solvent systems

The symbol ‘∞’ means the partition coefficient is too big that cannot be evaluated.