G6465b triple quadrupole uplc ms ms

This followed our previous method with suitable updates [19 (link)]. Freeze-dried powdered pepper fruit (20 mg) was homogenized in 1 mL extracting solution (ethanol: water = 1:1, V: V). The homogenate was shaken for 5 min before ultrasonication for 60 min and centrifugation at 10 000 g for 5 min at 4 °C. The supernatant was filtered through a 0.22-µm nylon filter before analysis using an Agilent G6465B triple quadrupole UPLC-MS/MS coupled with an HPLC reverse phase C18 column (Eclipse Plus C18 2.1 × 50 mm, 1.8 μm). The flow rate was 0.4 mL/min. The mobile phases A and B were acetonitrile and 0.1% formic acid in the water, respectively. The gradient elution was 5% A for 0 min, 100% A for 4 min, 5% A for 4.1 min, and 5% A for 5.3 min. The MS was performed using multiple reaction monitoring modes (MRM) and positive electrospray ionization. Additional file 1: Table S6 lists the specific instrumental parameters.

S. grosvenorii fruits (100 mg) were extracted with 1 mL of 60% ethanol in water [12 (link)]. The homogenate was subjected to ultrasound for 30 min and centrifuged at 12,000 rpm for 5 min. All supernatants were collected by repeating the operating method twice. The mixture was dried by blowing with nitrogen and maintained at a constant volume of 100 mL with 60% ethanol. The solutions were purged with 100 mg C18 and filtrated using a 0.22 μm nylon syringe. The Agilent G6465B triple quadrupole UPLC-MS/MS was equipped with an HPLC reverse phase C18 column (Eclipse Plus C18 2.1 × 50 mm, 1.8 μm). The flow rate was 0.4 mL/min and 5 µL injection volume. The mobile phases A and B consisted of acetonitrile and 0.1% formic ultrapure water, respectively. The instrument parameters of flavonoids and phenolic acid compounds are provided in Tables S2 and S3.