Hypesil gold column c18

80 μL of each plasma sample was added to 4 x chromatographic methanol, vortexed for 30 s, stood at 4°C for 1 h and then centrifuged at 12000 rpm for 10 min. The supernatant obtained by centrifugation was centrifuged at 12000 rpm for 10 min and 100 μL of the supernatant was added to the injection flask for detection. Meanwhile, 20 μL of each sample was taken as the quality control sample (QC).
Each sample was injected into the UPLC-Q Exactive-HF MS system equipped with electrospray ionization (ESI) ion source operated in both positive and negative-ion modes (Thermo Fisher, United States). Separation was carried out on hypesil gold column C18 (100 × 2.1 mm, 1.9 μm) (Thermo Fisher, GER), column temperature was set at 40°C; ESI + mode, use 0.1% formic acid as mobile phase A and methanol as mobile phase B; ESI- mode, use 5 mmol/L ammonium acetate (pH = 9.0) as mobile phase A and methanol as mobile phase B; Flow rate: 0.2 mL×min^-1. Gradient elution began at 98%A- 2%B for the first 0–12 min; at 12–14.1 min, it followed by 0%A- 100%B, and 98%A- 2%B for the final 14.1–17.0 min. Mass spectrometry conditions: capillary temperature, 320°C; spray voltage, 3.2 kV; sheath gas velocity, 40 arb; auxiliary gas flow rate, 10 arb; scanning range, m/z 100–1500.

Approximately 100 mg of each celery sample was transferred to a 1.5 mL centrifuge tube with 500 μL of extraction solution (methanol, water volume ratio of 4:1), followed by vortexing and incubation in an ice bath for 5 min, then centrifuging at 15,000× g for 20 min at 4 °C. Some supernatants were diluted to a final concentration of 53% methanol and analyzed by UHPLC-MS/MS. Samples of equal volume were taken from each experimental sample and mixed to obtain quality control samples (QC), inserted before, during and after the samples to test the repeatability of the experiment. The chromatographic and mass spectrometric conditions were as follows: a Hypesil Gold Column C18 (100 × 2.1 mm, 1.9 μm; Thermo Fisher Scientific, CA, USA) was used, the column temperature was set to 40 °C, the flow rate was 0.2 mL/min. In the positive ion mode, mobile phase A and mobile phase B components were 0.1% formic acid-aqueous and methanol, respectively. In the negative ion mode, mobile phase A and mobile phase B components were 5 mM ammonium acetate and methanol. The chromatographic gradient elution protocol consisted of: 0–12 min, 98% A, 2% B; 12–14.1 min, 100% B; 14.1–17 min, 98% A, 2% B. Mass spectrometry scan range 100–1500 m/z; ESI source settings: Spray Voltage: 3.2 kV; Aux Gasflow rate: 10 arb; Sheath gas flow rate: 40 arb; Capillary Temp: 320 °C; Polarity: positive, negative.

Instruments: Q Exactive™ HF-X (Thermo Fisher, Waltham, MA, USA), Vanquish UHPLC (Thermo Fisher, Waltham, MA, USA); Chromatographic columns: Hypesil Gold column (C18) (Thermo Fisher, Waltham, MA, USA). The column temperature was 40 °C, and the flow rate was 0.2 mL/min. In positive mode: mobile phase A was 0.1% formic acid (Thermo Fisher, USA); mobile phase B was methanol (Thermo Fisher, Waltham, MA, USA). In negative mode: mobile phase A was pH 9.0, 5 mM ammonium acetate (Thermo Fisher, Waltham, MA, USA); mobile phase B was methanol.
The scan range was m/z 100–1500, spray voltage was 3.5 kV; sheath gas flow rate was 35 psi; Aux gas flow rate is 10 L/min; Capillary temp was 320 °C; S-lens RF level was 60; Aux gas heater temp was 350 °C. Mobile phase gradient settings: Time: 0–1.5 min, A: 98%, B: 2%; Time: 3 min, A: 15%, B: 85%; Time: 10 min, A: 0, B: 100%; Time: 10.1–12 min, A: 98%, B: 2%.