Empower software version 3

UPLC analysis was performed using a Waters ACQUITY UPLCTM system (Waters Corporation) equipped with a quaternary solvent manager system, autosampler and a PDA detector. The samples were eluted using a Waters ACQUITYUPLC HSS T3 column (100 × 2.1 mm, 1.7 μm). The column and autosampler temperatures were maintained at 40 °C and 25 °C, respectively. A mobile phase containing water (A) and ACN (B) was used with the optimized gradient program as follows: 10%–25% B (0–0.8 min), 25%–35% B (0.8–2.4 min), 35%–95% B (2.4–2.8 min), 95% B (2.8–4 min). The flow rate was set at 0.5 mL/min. The sample injection volume was 1 μL. The wavelength of PDA detector was set to 260 nm. Empower software Version 3.0 (Waters Corporation) was used for system control and data acquisition [32 (link)].

The chemical composition of the IGF was analyzed using UPLC. Crude extracts were dissolved in methanol to produce a final concentration of 0.5 mg/mL. The standards (verproside, catalposide, and amphicoside) were dissolved in methanol to produce a final concentration of 0.1 mg/mL, respectively. Our UPLC-photodiode array (PDA) used a Waters Acquity System (Waters Co., Milford, MA, USA) that consisted of a PDA, binary autosampler, online degasser, and column oven. The UPLC system was fitted with an Acquity UPLC HSS T3 column (100 × 2.1 mm, 1.7 μm; Waters Co.). The column and autosampler temperatures were maintained at 40°C and 25°C, respectively. The mobile phases A and B were ultrapure water and acetonitrile, respectively. The following gradient profile was used: 10–25% B (0–0.8 min), 25–35% B (0.8–2.4 min), 35–95% B (2.4–2.8 min), 95% B (2.8–4 min), and 95–100% B (4–8 min). The flow rate was 0.5 mL/min and the injection volume was 1.0 µL. Compounds were identified by a comparison of the retention times between samples and standards. The PDA detection was conducted at 260 nm. Levels of the IGF were quantitated using Empower software version 3.0 (Waters) [10 (link), 16 (link)].

A Waters Alliance e2695 HPLC system (Waters Corp., Milford, MA, USA) equipped with a pump, degasser, column oven, auto sample injector, and photodiode array (PDA) detector (#2998, Waters Corp. Milford, MA, USA) was used in the quantitative analysis and Empower software (version 3; Waters Corp, Milford, MA, USA) was used to data processing. The chromatographic separation of the seven marker compounds was performed at 30 °C using a Gemini C-18 analytical column (250 × 4.6 mm, 5 μm; Phenomenex, Torrance, CA, USA) with a gradient solvent system of 1.0% (v/v) aqueous acetic acid (A) and acetonitrile (B). The elution conditions were as follows: 12–42% B for 0–25 min, 42–52% B for 25–30 min, 52–65% B for 30–55 min, 65–100% B for 55–56 min, and 100% B for 56–63 min. The flow rate and injection volume were 1.0 mL/min and 10 μL, respectively. The wavelength range of the PDA detector was 190 nm to 400 nm.