Standard aloe-emodi, rhein, emodin, chrysophanol, and physcion were obtained from Shanghai R&D Center for Standardization of Traditional Chinese Medicines. High-performance liquid chromatography (HPLC)-ultrapure water, analytical-grade methanol, NaHCO3, and phosphoric acid were purchased from Sangon Biotech, Ltd. (Shanghai, China).
The metabolites of R. palmatum were measured according to the methods of Chen et al. [32 (link)]; the dried root of different treatments (five replicates) was pulverized and sieved by using a mesh (300 μm). An amount of 0.15 g of powder was precisely weighed, 10 mL 0.1% (w/v) NaHCO3 solution was added, and ultrasonic treatment was performed (30–40 °C, 700 W) for 20 min. Then, 40 mL of methanol was added and treated using ultrasound for 50 min. The sample solution was filtered by a Millipore filter unit (0.22 μm), and then 10 μL of sample solution was injected into the HPLC for measurement.
HPLC system and parameters were set according to the methods of Chen et al. [33 (link)]; the metabolites content of samples was determined by HPLC (Waters E2695, Milford, MA, USA) using C18 (4.6 × 250 mm, 5.0 μm) at 30 °C. The mobile phase of HPLC was methanol −0.1% phosphoric acid (80:20); the flow rate was 1 mL/min. The detection wavelength was 254 nm.
The metabolites of R. palmatum were measured according to the methods of Chen et al. [32 (link)]; the dried root of different treatments (five replicates) was pulverized and sieved by using a mesh (300 μm). An amount of 0.15 g of powder was precisely weighed, 10 mL 0.1% (w/v) NaHCO3 solution was added, and ultrasonic treatment was performed (30–40 °C, 700 W) for 20 min. Then, 40 mL of methanol was added and treated using ultrasound for 50 min. The sample solution was filtered by a Millipore filter unit (0.22 μm), and then 10 μL of sample solution was injected into the HPLC for measurement.
HPLC system and parameters were set according to the methods of Chen et al. [33 (link)]; the metabolites content of samples was determined by HPLC (Waters E2695, Milford, MA, USA) using C18 (4.6 × 250 mm, 5.0 μm) at 30 °C. The mobile phase of HPLC was methanol −0.1% phosphoric acid (80:20); the flow rate was 1 mL/min. The detection wavelength was 254 nm.
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