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UPLC-MS/MS
The analytical conditions were as follows, UPLC: column, C18 (1.8 μm, 2.1 mm × 100 mm); the mobile phase was consisted of solvent A, pure water with 0.1% formic acid, and solvent B, acetonitrile. Sample measurements were performed with a gradient program that employed the starting conditions of 95% A, 5% B. Within 9 min, a linear gradient to 5% A, 95% B was programmed, and a composition of 5% A, 95% B was kept for 1min. Subsequently, a composition of 95% A, 5.0% B was adjusted within 1.10 min and kept for 2.9 min. The column oven was set to 40 ℃. The injection volume was 4 μL. The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 550 ℃; ion spray voltage (IS) 5500 V (positive ion mode)/-4500 V (negative ion mode); ion source gas I (GSI), gas II(GSII), curtain gas (CUR) were set at 50, 60, and 30.0 psi, respectively; the collision gas (CAD) was high. Instrument tuning and mass calibration were performed with 10 and 100 μM polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to 5 psi. DP and CE for individual MRM transitions were done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period.
References：
[1] Xiao Y , Li K , Zhang H ,et al.The profile of buckwheat tannins based on widely targeted metabolome analysis and pharmacokinetic study of ellagitannin metabolite urolithin A[J].LWT, 2022, 156:113069-.DOI:10.1016/j.lwt.2022.113069.
