Sr 3000 solvent rack

Quantitative analysis and fingerprint analysis were performed using UltiMate 3000 HPLC instrument (Thermo-Fisher, USA) with a DAD 3000 detector, a ternary pump of SR3000 Solvent Rack, a WPS-3000SL autosampler, a TCC-3000SD column temperature controller, and a workstation of Chromeleon 7.2. The compounds separation was performed on a Thermo Hypersil Gold C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of 0.1% phosphoric acid (solvent A) and Acetonitrile (solvent B) at a flow rate of 1 mL/min, which followed a gradient program of 0–20 min, 18–35% B; 20–30 min, 35–48% B; 30–35 min. The column temperature was maintained at 30 °C while the autosampler temperature was maintained at 4 °C with 10 μL of sample solution being injected. Both the detection wavelength of DAD was monitored at 235 nm.

All of the HPLC analyses were performed with a Dionex U-3000 series equipped with a SR-3000 Solvent Rack, a LPG-3400SDN Quaternary Pump, a WPS-3000SL Auto sampler, a TCC-3000RS Column compartment, a DAD-3000RS detector, and a Chromeleon 7 chromatography workstation (Thermo Fisher Scientific, Waltham, MA, USA). An Agilent ZORBAX Extend-C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of acetonitrile (A) and water (B). The gradient program was developed as follows: 15–28% A for 0–22 min, 28–37% A for 22–35 min, 37–50% A for 35–45 min, and 50–75% A for 45–60 min. The flow rate was maintained at 1.0 mL/min and the column temperature at 30 °C. The injection volume was 10 μL and the detective wavelength was selected at 205 nm.

HPLC analysis was conducted according to the method described previously [20 (link)]. Briefly, the HPLC analysis was performed using an UltiMate 3000 HPLC analysis system (Thermo-Fisher, Waltham, MA, USA) with a DAD 3000 detector, ternary pump of SR3000 Solvent Rack, WPS-3000SL autosampler, TCC-3000SD column temperature controller, and Chromeleon 7.2. Separation and was achieved in a C18 reversed-phase column (Hypersil Gold, particle size 5 μm, 250 mm × 4.6 mm, Thermo Scientific). The mobile phase contained acetonitrile (solvent A) and 0.1% phosphoric acid aqueous solution (solvent B). The detailed gradient elution program was as follows: 0–25 min, 25–50% A; 25–30 min, 50–99% B; and 30–50 min, 99–99% B; and HPLC profiling was performed at 30°C at a constant flow rate of 1.0 mL/min. All samples were injected into the system at equal volumes of 10 μL. Ganoderic acid A, ganoderic acid G, lucidenic acid A, and ergosterol were used as standards, and an analysis wavelength of 254 nm was selected.