Acquity uplc liquid chromatography

Acquity™ UPLC liquid chromatography (Waters, USA); Synapt™ G2-Si mass spectrometry (Waters, USA); Masslynx v4.1 workstation (Waters, USA); Ultrasonic cleaner (KQ-250 DB, Kunshan Ultrasonic Instrument Co., Ltd., China); Table centrifuge (sorvall ST 16R, Thermo Scientific, USA); Thermo Scientific 995 ultra low temperature refrigerator (Thermo Scientific, USA).

Chromatograph condition: Acquity™ UPLC liquid chromatography (Waters, USA); chromatographic column used was an ACQUITY HSS T3 chromatographic column (100 mm × 2.1 mm i.d., 1.8 μm. Waters, USA); the column temperature is set at 40°C; the sample bin temperature is set at 4°C; meanwhile mobile phase for gradient elution consist of phase A (acetonitrile containing 0.1% formic acid) and phase B (water with 0.1% formic acid). The gradient was as follows: 0–1 min, 1%A; 1–6 min, 1–30%A; 6–8 min, 30–70%A; 8–9 min, 70–99%A; 9–10 min, 99%A; 10–10.1 min, 99–1%A; 10.1–14 min, 1%A. In addition, equal volume of each sample was mixed into a quality control sample (QC) for monitoring the stability of the instrument in the experiment, and one QC solution was run every ten samples, which were crucial for optimization during metabolomic analytical method development.

In this study, the instruments used included Acquity™ UPLC liquid chromatography (Waters, United States), Synapt™ G2-Si mass spectrometry (Waters, United States), Masslynx v4.1 workstation (Waters, United States), PRO-MRRM-8 small animal energy metabolism monitoring system (Sable Systems International, United States), KQ-250 DB ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd., China), Sorvall ST 16R table centrifuge (Thermo Scientific, United States), and Thermo Scientific 995 ultra-low temperature refrigerator (Thermo Scientific, United States).