NODEMetabolomicsJian Wang4000 QTRAPhttps://www.biosino.org/node/experiment/detail/OEX000211161['seeds']Samples to be analyzed were freeze-dried with a vacuum freeze dryer and ground into powder using a grinder (30 Hz, 1.5 min). Then, 100 mg of sample was dissolved in 1.2 ml 70% MeOH, and incubated with vortexing every 30 min for a total of six rounds of vortex-ing. Afterwards, samples were placed in the refrigerator at 4 °C overnight. The next day, the samples were centrifuged at 12000 rpm for 10 min and the supernatant extracted and filtered by a micropore filter (0.22 μM pore size). The samples were then transferred into sample bottles for subsequent UPLC-MS/MS analysis. Ultra-performance liquid chromatography (SHIMADZU Nexera X2, https://www.shimadzu.com.cn/) coupled with tandem mass spectrometry (Applied Bio-systems 4500 QTRAP，http://www.appliedbiosystems.com.cn/) were used to perform the widely untargeted metabolic analysis of samples. Chromatographic separation was per-formed on an Agilent SB-C18 column (1.8 µm，2.1 mm*100 mm). The mobile phases consisted of ultrapure water with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B). The gradient elution program was set as follows: 95:5 A/B at 0 min, 5:95 A/B at 9 min, 5:95 A/B at 10 min, 95:5 A/B at 11.10 min, and 95:5 A/B at 14 min. The flow rate was 0.35 ml/min, the column temperature was 40 °C, and the injection volume was 4 μl. The linear ion trap and QQQ scanning were performed using an AB4500 Q TRAP MS / MS system. MS acquisition utilized both positive and negative ion modes. The parameters for electrospray ionization (ESI) were as follows: source temperature, 550 °C; ion spray volt-age, 5500 V (+) / - 4500 V (-). QQQ scanning employed multiple reaction monitoring.['Glycine max']NODEfalseOEX_Jian_2211281141A widely untargeted metabolic analysis was performed on ZX4 and ZX12 seeds col-lected at 20, 30, and 40 DAF, with three biological replicates.2022-11-282022-11-28OEX00021116OEP00003774