The LC-20AD UPLC system (Shimadzu Co., Kyoto, Japan) consisted of a LCMS-8030 (Shimadzu Co., Kyoto, Japan) triple quadrupole mass spectrometer equipped with LC-20AD pump (Shimadzu Co., Kyoto, Japan), CTO-20A column oven (Shimadzu Co., Kyoto, Japan), DGU-20A3 online degasser (Shimadzu Co., Kyoto, Japan), and SIL-20ACXR autosampler (Shimadzu Co., Kyoto, Japan). Furthermore, the tandem quadrupole mass spectrometer equipped with electrospray ionization (ESI) turbo ion interface was used with the following parameters: interface voltage (kV): 4.5; DL temperature (°C): 250; heat block temperature (°C): 400; nebulizing gas flow (L/min): 3; and drying gas flow (L/min): 15 and nitrogen was used in all cases.
A C18 column (ACQUITY BEH, 100 mm × 2.1 mm i.d., particle size 1.7 mm, Waters, Ireland) was used for UPLC separation. The mobile phase consisted of (A) methanol with 0.1% formic acid and (B) 5 mM NH4OAC. The gradient elution program of mobile phase was as follows: 0-1 min: 25–70% A; 1–3.5 min: 70–90% A; 3.5–7.5 min: 90-90% A; 7.5–8.5 min: 90–25% A; and 8.5–13 min: 25-25% A, v/v. The flow rate for the mobile phase was set at 0.2 mL/min and the analytical volume was 5 μL of JWXYS extract.
A C18 column (ACQUITY BEH, 100 mm × 2.1 mm i.d., particle size 1.7 mm, Waters, Ireland) was used for UPLC separation. The mobile phase consisted of (A) methanol with 0.1% formic acid and (B) 5 mM NH4OAC. The gradient elution program of mobile phase was as follows: 0-1 min: 25–70% A; 1–3.5 min: 70–90% A; 3.5–7.5 min: 90-90% A; 7.5–8.5 min: 90–25% A; and 8.5–13 min: 25-25% A, v/v. The flow rate for the mobile phase was set at 0.2 mL/min and the analytical volume was 5 μL of JWXYS extract.