Zobax sb c18 column

The EFE profile was obtained in a high-performance liquid chromatography (HPLC) with diode array detector (DAD) system (Agilent 1200). HPLC separation was conducted with an Agilent Zobax SB-C18 column (4.6 mm × 250 mm) maintained at 30°C, a flow rate of 1 ml/min with an injection volume of 10 μl, and detection wavelength was 270 nm. The chromatographic conditions were as follows: the column temperature of 30°C; the detection wavelength used at 270 nm; mobile phase A, acetonitrile; mobile phase B, distilled water. The elution program was set as follows: 0 min–30 min and 24–26% A (v/v); 30 min–35 min and 26–35% A (v/v); 35 min–50 min and 35–50% A (v/v); and 50 min–55 min and 50–52% A (v/v).
EFE was dissolved by ultrasonic with distilled water, separated by C18A31 column chromatography and eluted with a gradient of 25% acetonitrile and 50% acetonitrile to generate two fragments, and analyzed by high-performance liquid chromatography. The elution program was set as follows: 0 min–30 min and 24%–26% A (v/v); 30 min–31 min and 26%–45% A (v/v); 31 min–45 min and 45%–47% A (v/v); 45 min–46 min and 47%–95% A (v/v); and 46 min–60 min and 95%–24% A (v/v). Other conditions are the same as earlier mentioned.
All authentic samples (purity ≥ 95.0%) were purchased from Shanghai Standard Technical Service Co., Ltd. (Shanghai, China).

TBE-300C HSCCC equipment was used with three multilayer columns of a total of 300 mL (the diameter of the PTFE tube was 2.6 mm) and a 20 mL sample loop ( Tauto Biotech, Shanghai, China). An LC3050N integrated system was used to contact the HSCCC equipment, composed with a P3050N gradient pump and a UV3050N monitor (Beijing Tong Heng Innovation Technology Co., Ltd., Beijing, China). A DC-0506 low-constant-temperature bath was used to control the temperature at 30 °C. An Agilent 1120 with a Zobax SB C₁₈ column (5 μm, 250 mm × 4.6 mm, i.d.) was used for the sample detection process (Agilent Technologies, Santa Clara, CA, USA).