1260 series hplc instrument

Characterization of the components of JBT was performed by a 1260 series HPLC instrument (Agilent, Waldbronn, Germany) connected to an Agilent 6530 Q-TOF mass spectrometer (Agilent Corp., USA) equipped with Dual Agilent Jet Stream Electrospray Ionization (Dual AJS ESI). The operating parameters were optimized in both positive and negative modes, as follows: capillary voltage, 3,500 V for ESI mode and 4000 V for ESI+ mode; nozzle voltage, 500 V; fragmentor, 110 V; nebulizer, 45 psi; drying gas temperature, 300°C; drying gas flow rate, 6 L/min; sheath gas temperature, 320°C; and sheath gas flow rate, 12 L/min. Separation of compounds in JBT was carried out on an ACE Excel 3 Super C18 column (100 mm × 2.1 mm; Advanced Chromatography Technologies Ltd., Aberdeen, Scotland). The mobile phase was composed of solvent A (water containing 0.1% formic acid) and solvent B (acetonitrile containing 0.1% formic acid), with a flow rate of 0.35 ml/min. Gradient elution was performed as follows: 0–30 min, 95%–23% A; 30–40 min, 23%–15% A; 40–45 min, 15%–10% A; 45–50 min: 10%–5% A; 50–55 min, 5% A; 55–56 min: 5%–95% A; and 56–66 min, 95% A. The column temperature was maintained at 40°C. System operations and data analysis were conducted on Masshunter Workstation software (Agilent Technologies, USA).

An Agilent 1260 series HPLC instrument (Santa Clara, CA, USA) equipped with an Agilent DAD detector (All Tech, Lexington, KY, USA) was used to obtain HPLC fingerprints. A Thermo C₁₈ column (250 mm × 4.6 mm, i.d., Agilent) was used for analyses. For separation, a column temperature of 30°C, a flow rate of 1.0 mL/min, and an injection volume of 10 μL were used.
The optimized elution conditions were as follows: (C) acetonitrile and (D) 0.3% formic acid in water were used as the mobile phase. A linear gradient of 0%–7% (C) (0–15 min), 7%–20% (C) (15–20 min), and 20%–43% (C) (20–40 min) with wavelength conditions of 273 nm (0–20 min), 360 nm (21–30 min), and 270 nm (31–40 min) was applied.