Metabolomics raw data

Description

Metabolomics raw data

Steps to reproduce

1. Extraction of Liver Tissue Metabolites Liver samples (approximately 50 ± 5 mg per mouse, n=6 per group) were collected and placed individually into 2 mL centrifuge tubes. A total of 800 μL methanol-water extract was added to each tube. Samples were ground in a cryogenic tissue homogenizer at −10 °C, 50 Hz for 6 min, followed by low-temperature ultrasonic extraction at 4 °C, 40 kHz for 30 min. Samples were then incubated at −20 °C for 30 min and centrifuged at 13,000 g for 15 min at 4 °C. The supernatant was collected for subsequent analysis. All individual sample data were included in statistical comparisons between groups. 2. LC-MS/MS Analysis Liver metabolites were analyzed using a UHPLC-Triple TOF 6600 system (AB SCIEX, USA). A 10 μL aliquot of supernatant was separated on an ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm i.d., 1.8 μm; Waters, USA). The mobile phase flow rate was 0.40 mL/min, and column temperature was maintained at 45 °C. Mobile phase A consisted of 95% water and 5% acetonitrile; mobile phase B consisted of 47.5% acetonitrile, 47.5% isopropanol, and 5% water, both containing 0.1% formic acid. The gradient elution program was as follows: 0–0.2 min, 0% B; 0.2–3 min, 25% B; 3–9 min, 25% to 100% B; 9–10 min, 100% B; 10–10.1 min, 100% to 0% B; 10.1–12 min, 0% B. Ion spray voltage was 5500 V (positive) and −4500 V (negative). The mass scan range was 50–1200 m/z. Sheath gas flow was 50 arb, auxiliary heating gas flow was 13 arb, ion source temperature was 450 °C, and collision energy was set at 20, 40, and 60 V. 3. Metabolomics Data Analysis Raw data were processed using Progenesis QI software (Waters, USA). Metabolites were identified by matching MS and MS/MS spectra against HMDB, Metlin, and the Majorbio custom database. The data matrix was uploaded to the Majorbio cloud platform (cloud.majorbio.com) for preprocessing. PLS-DA and OPLS-DA were performed using the ropls package (v1.6.2) in R, with 7-fold cross-validation to validate model stability. Differential metabolites were screened with VIP > 1 and p < 0.05. KEGG annotation and pathway enrichment analysis were conducted using the Python package “scipy.stats” and Fisher’s exact test to identify key biological pathways.

Institutions

• Xinjiang Technical Institute of Physics & Chemistry

  Xinjiang, Ürümqi

Categories

Licence