1290 series liquid chromatography system

The composition and content of each anthocyanin was determined using HPLC-DAD-MS/MS with a 1290 Series liquid chromatography system (Agilent Technologies Inc., Palo Alto, CA, USA) as previously published [28 (link),29 (link),30 (link)].

Lovastatin was used as the internal standard (IS) for the analysis of simvastatin. The analysis was conducted with Agilent 1290 series liquid chromatography system and an Agilent 6460 triple-quadrupole mass spectrometer (Palo Alto, CA, USA). Water and acetonitrile were used as the mobile phase with a flow rate of 0.3 mL/min and an analysis time of 4 min. The m/z ratios of precursor-to-product ion reactions for simvastatin and IS were 419.32/199.15 and 405.3/199.15, respectively.

The determination of HCPT was performed on an Agilent 1290 series liquid chromatography system and an Agilent 6470 triple-quadruple mass spectrometer (Palo Alto, CA). The HPLC/MS conditions and sample preparation were basically according to a validated HPLC method described before (Zhang et al. 2019 ). The chromatographic analysis of HCPT was performed on a Waters X-Bridge C18 column (3.0 × 100 mm, i.d.; 3.5 μm, Waters Corporation, Milford, MA ) at room temperature (25 °C). The mobile phase was water (containing 0.1% formic acid) and acetonitrile (30:70, v: v) with isocratic elution at a flow rate of 0.2 mL/min, and the analysis time was 4 min.
The mass scan mode was positive MRM mode. The precursor ion and product ion are m/z 365.20→321.10 for HCPT and m/z 349.25→305.15 for IS. The collision energy for HCPT and IS were 30 and 20 eV, respectively. The MS/MS conditions were optimized as follows: fragmentor, 110 V; capillary voltage, 3.5 kV; Nozzle voltage, 500 V; nebulizer gas pressure (N₂), 40 psig; drying gas flow (N₂), 10 L/min; gas temperature, 350 °C; sheath gas temperature, 400 °C; sheath gas flow, 11 L/min.

The determination of warfarin was performed using an Agilent 1290 series liquid chromatography system and an Agilent 6470 triple-quadruple mass spectrometer (Palo Alto, CA, USA). The chromatographic analysis of warfarin was performed on a Waters X-bridge C18 column (3.0 × 100 mm, i.d.; 3.5 μm) at room temperature. The mobile phase was water (containing 0.1% formic acid) and acetonitrile (35:65, v:v) at a flow rate of 0.3 mL/min. The mass scan mode was negative MRM mode. The precursor ion and product ion are m/z 307.0 → 250.1 for warfarin, and m/z 301.0 → 121.1 for quercetin (internal standard), respectively. The collision energy for warfarin and quercetin were 20 and 25 ev, respectively. The MS/MS conditions were optimized as follows: fragmentor, 110 V; capillary voltage, 3.5 kV; nozzle voltage, 500 V; nebulizer gas pressure (N₂), 30 psig; drying gas flow (N₂), 10 L/min; gas temperature, 350 °C; sheath gas temperature, 400 °C; and sheath gas flow, 11 L/min.

The chromatographic analysis was performed with the Agilent 1290 series liquid chromatography system and an Agilent 6470 triple-quadruple mass spectrometer (Palo Alto, CA, USA) with the Waters X-Bridge C18 column (3.0 × 100 mm, i.d.; 3.5 μm, USA). The mobile phase was water (containing 0.1% formic acid) and acetonitrile (30:70, v: v) with isocratic elution at a flow rate of 0.2 mL/min, and the analysis time was 4 min.
The quantification was conducted in a multiple reaction monitoring (MRM) mode with the m/z of 323.1 → 119.0 for bavachin and 255.1 → 119.0 for liquiritigenin as the internal standard (IS). The collision energy for bavachin and IS were 30 and 20 eV, respectively. The MS/MS conditions were optimized as follows: fragmentor, 110 V; capillary voltage, 3.5 kV; Nozzle voltage, 500 V; nebulizer gas pressure (N₂), 40 psig; drying gas flow (N₂), 10 L/min; gas temperature, 350 °C; sheath gas temperature, 400 °C; sheath gas flow, 11 L/min.