1260 infinity 2 high performance liquid chromatograph

The following instruments were used: a Milli-Q water purifier (Millipore Company, United States); a CT15RE 4°C centrifuge (Hitachi, Japan); a G-560E scroll oscillator (Scientific Industries, United States); the Mettler Toledo ME104E Electronic Balance (China Mettler Toledo Instrument Co., LTD.); a BPH-9162 high-temperature drying oven (Shanghai Heng Scientific Instrument Co., Ltd., China); an ultra-high performance liquid-phase system (Vanquish, Thermo Fisher Scientific); a high-resolution mass spectrometer (Q Exactive Focus, Thermo Fisher Scientific); the 1260 Infinity II high-performance liquid chromatograph (Agilent Corporation, United States); and the EASY-nLC 1000 nA upgraded liquid chromatograph (Thermo Fisher Scientific, United States).

HPLC was utilized to identify the potential active substances of DTYMT. In brief, 10 mL DTYMT dilution solution (DTYMT: water = 1:9) and stands were respectively injected into the 1260 Infinity II High Performance Liquid Chromatograph equipped with a diode array detector working in the range of 200–400 nm, column temperature chamber and autosampler (Agilent Technology, CA, USA). A Kromasil 100-5-C18 column (4.6 mm*250 mm, 5 μm, Nouryon, Sweden）was used for the separation of the components and the flow column temperature was 30 °C. The mobile phase consisted of methanol (A) and 0.02% phosphoric acid water (B). The gradient elution programme was used as follows: 0–5 min,15–20%A; 5–30 min, 20–48%A; 30–40 min, 48–57%A; 40–45 min, 57-36%A; 45–55 min, 78–80%A and the re-equilibration time of gradient elution was 5 min. The flow rate was 1 mL/min, and the injection volume was set at 10 μL. Peaks were monitored at 224 nm.