7890b 7000d gc ms ms platform

Winter buds collected from 60 DABB and 67 DABB were sent to Metware Biotechnology (Wuhan, China) to detect endogenous hormone concentrations. The content of auxin, cytokinin, and gibberellin was measured using the AB Sciex QTRAP 6500 LC-MS/MS platform with internal standard methods, while the ACC content was determined using the Agilent 7890B-7000D GC-MS/MS platform. Three biological replications were performed.

The colon contents samples were delivered to Metware Biotech (Wuhan, China) to employ a widely targeted metabolomics assay. The BA contents were detected using the AB Sciex QTRAP 6500 LC-MS/MS platform. A total of 20 mg of sample was extracted with methanol after grinding, and then added to the internal standard for quantitation. After protein precipitation and centrifugation, the supernatant was collected. The extracts were evaporated to dryness and reconstituted in methanol, followed by liquid chromatography–mass spectrometry analysis. Analyst (Version 1.6.3) was utilized for data collection, and Multiquant (Version 3.0.3) was utilized for metabolite quantification.
The SCFA contents were detected using the Agilent 7890B-7000D GC-MS/MS platform. A total of 20 mg of sample was weighed, dissolved in phosphoric acid solution, homogenized under ultrasonication, and centrifuged to acquire a supernatant. Following being added with tert-butyl methyl ether containing internal standard, the mixture was vortexed, ultrasonicated, and centrifuged again to collect the supernatant for gas chromatography–mass spectrometry analysis.
Orthogonal partial least squares discriminant analysis (OPLS-DA) was performed to analyze the data to explore the differences in metabolic profiles between each group.