Acquity uplc 1 class xevo tq s micro system

Metabolic analysis of the PxJHE protein was investigated by UPLC‐MS/MS analysis using the ACQUITY UPLC I‐Class/Xevo TQ‐S micro System (Waters) equipped with an ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 µm particle size), and JH II standard (SciTech; 78%) were used as metabolic substrate. 0.18 µg recombinant PxJHE protein was added to 50 µL reaction volume containing 20 µg JH II. After incubation at 30 °C for 5 min, the reaction was stopped by adding 50 µL methanol. Subsequently, 100 µL of the sample was transferred to a brown loading bottle, and the product was further detected by the ACQUITY UPLC I‐Class/Xevo TQ‐S micro System (Waters). The column temperature was set to 40 °C. A methanol/water (0.1% formic acid) gradient elution was carried out at a flow rate of 0.2 mL min⁻¹ as follows: 0–2 min (70% methanol), 2–4 min (85% methanol), maintained for another 2 min, 6–8 min (70% methanol) with an injection volume of 10 µL. The entire run lasted 9 min, including a 1 min equilibration step. Data acquisition and analysis were performed using the MassLynx V4.1 software (Waters) as mentioned above.