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databases/metabolights/studies/public/MTBLS14033 All statistical analyses were performed with SPSS software v26.0 (IBM Corporation, Chicago, IL, United States).One-way analysis of variance (ANOVA) was used to compare inter-group differences at the same experimental time point, with LSD employed for post-hoc tests. For continuous variables that did not conform to a normal distribution, the Kruskal-Wallis H test was used to compare inter-group differences at the same experimental time point.MetaboLightsPublicLiquid Chromatography MS - alternating - hilic Chromatographic separation was performed on an UltiMate 3000 system using a C18 column maintained at 35°C. The mobile phases consisted of (A) 100% acetonitrile and (B) water containing 0.1% formic acid. A gradient elution program was applied at a flow rate of 0.2 mL/min as follows: 5% A to 50% A from 0 to 7 min; 50% A to 75% A from 7 to 8 min; 75% A to 80% A from 8 to 9 min; 80% A to 90% A from 9 to 11 min; 90% A to 95% A from 11 to 15 min; finally, 95% A was held from 15 to 20 min.Cognitive Improvement and Microbiota-Gut-Brain Axis Regulation by Lycium barbarum polysaccharides and glycopeptide in Laboratory-Kenneled Poodles.South China Agricultural UniversityYan Haoranserumfecesmass spectrometry assay I. Pretreatment of Serum Samples1. Sample ExtractionFollowing thawing, samples were vortexed for 2 min to ensure thorough homogenization. A 200 µL aliquot of each serum sample was transferred to a sterile, nuclease-free EP tube, followed by the addition of 800 µL of ice-cold methanol (chromatography grade) for protein precipitation. The mixture was vortexed for 2 min and subsequently centrifuged at 14,500 rpm, 4 °C for 15 min. A 1000 µL volume of the supernatant was collected and subjected to vacuum centrifugation for 3 hours. If complete dryness was achieved via vacuum centrifugation, nitrogen stream evaporation was omitted. Otherwise, the sample was dried under a gentle nitrogen stream at room temperature. The dried residue was stored at -80 °C. For the initial analysis, residues from 120 samples were selected for reconstitution, while the remaining samples continued to be stored for subsequent batch processing.For reconstitution, the dried residue was resuspended in 200 µL of a methanol:water (1:1, v/v) solution (chromatography grade), vortexed for 2 min, and sonicated in an ice bath for 10-15 min. The mixture was then centrifuged at 14,500 rpm, 4 °C for 15 min. The resulting supernatant was transferred to an autosampler vial (equipped with an insert) and stored at -80 °C pending analysis.2. Preparation of Quality Control (QC) SamplesFor each batch of serum samples, a pooled QC sample was prepared. Briefly, a 50 µL aliquot was taken from each of 48 individual samples, resulting in a total pooled volume of 2.4 mL. The pool was mixed thoroughly and aliquoted into 2-3 tubes. These aliquots then underwent the identical extraction and processing procedure described in Section 1 to generate the QC samples. The prepared QC samples were stored at -80 °C pending analysis.II. Pretreatment of Fecal Samples1. Fecal Sample PreprocessingThe frozen fecal samples were thawed at 4 °C. Approximately 60 mg of each sample was weighed into a 2 mL round-bottom microcentrifuge tube suitable for bead homogenization. Subsequently, 600 µL of methanol (chromatography grade) was added for metabolite extraction, and the mixture was homogenized using beads. The homogenate was subjected to low-temperature ultrasonication for 10 min, followed by incubation at -20 °C for 30 min. The samples were then centrifuged at 14,500 rpm, 4 °C for 15 min. A 200 µL aliquot of the supernatant was transferred and dried using vacuum centrifugation.The dried residue was reconstituted in 200 µL of a methanol:water (1:1, v/v) solution (chromatography grade), vortexed for 2 min, and sonicated in an ice bath for 10-15 min. This was followed by centrifugation at 14,500 rpm, 4 °C for 15 min. The resulting supernatant was transferred to an autosampler vial (equipped with an insert) and stored at -80 °C pending analysis.2. Preparation of Quality Control (QC) Samples For each batch of fecal samples, a pooled QC sample was prepared. A 60 mg aliquot was taken from each of 48 individual samples, combined, and thoroughly mixed to yield a total pooled mass of approximately 3 g. This pooled mixture was then aliquoted into 2-3 tubes. These aliquots underwent the identical preprocessing procedure described in Section 1 to generate the QC samples. The prepared QC samples were stored at -80 °C pending analysis.Canis lupus familiarishttps://www.ebi.ac.uk/metabolights/MTBLS14033Zhang Miaomiao. South China Agricultural University. No.483 Wushan Road, Tianhe District, Guangzhou, China. zhangmiao7839@163.com.Haoran Yan. South China Agricultural University. No.483 Wushan Road, Tianhe District, Guangzhou, China. 13680730396@136.com.Zhang Lingna. South China Agricultural University. No.483 Wushan Road, Tianhe District, Guangzhou, China. lingna.zhang@scau.edu.cn. Data processing was performed using Compound Discoverer 3.3 SP3 software (Thermo Fisher Scientific). This software was utilized to automatically process the raw data, and metabolite identification was conducted by searching against the mzCloud and mzVault mass spectral libraries. Targeted metabolomics was applied to quantify the metabolites involved in the Trp metabolism, Standard compounds were dissolved with methanol–water (1:1) and then diluted in a gradient to a standard solution (i.e., 1000, 500, 100, 10, 1, 0.5, 0.1, 0.01, 0.001 ng/mL). The standard solution was further analyzed to build standard curves using UPLC-Orbitrap-MS/MS. Finally, the raw data were imported into the Xcalibur software (version 4.3) to quantify the concentration of metabolites according to their standard curves.TreatmentSex13680730396@163.comAfter an 8-hour fast, 5 mL of blood was collected from the forelimb vein of each dog into blood collection tubes on Day 42. The tubes were tilted and allowed to stand for 30 minutes, then centrifuged at 3500 rpm for 15 minutes at room temperature. The supernatant was aliquoted into microcentrifuge tubes and stored at -80°C for further analysis.  On Day 42, fresh feces excreted within 10 minutes were collected. Using a fecal collector, uncontaminated samples from the interior portion of the feces were obtained. The samples were aliquoted into cryovials and stored at -80°C for further analysis.MetabolomicsMetabolomeDoglycium barbarum polysaccharideLycium barbarum fruit extractMetabolomeDoglycium barbarum polysaccharideLycium barbarum fruit extract Chromatographic separation was performed on an UltiMate 3000 system using a C18 column maintained at 35°C. The mobile phases consisted of (A) 100% acetonitrile and (B) water containing 0.1% formic acid. A gradient elution program was applied at a flow rate of 0.2 mL/min as follows: 5 % A to 50 % A from 0 to 10 min; 50 % A to 75 % A from 7 to 8 min; 75 % A to 80 % A from 8 to 9 min; 80 % A to 90 % A from 9 to 11 min; 90 % A to 95 % A from 11 to 15 min; finally, 95 % A was held from 15 to 20 min.Mass spectrometric detection was carried out on a Q-Exactive Focus instrument equipped with an electrospray ionization (ESI) source. The key source parameters were set as follows: spray voltage, +3500 V (positive mode) and -4000 V (negative mode); vaporizer temperature, 300 °C; sheath gas pressure, 40 arbitrary units (arb.); auxiliary gas pressure, 10 arb.; and capillary temperature, 320 °C.Two acquisition modes were employed:(1) Full scan/dd-MS² mode: Full scans were acquired at a resolution of 35,000 with an in-source collision-induced dissociation (CID) energy of 0 eV. Data-dependent MS² scans were triggered at a resolution of 17,500 using stepped normalized collision energies (NCE) of 10, 30, and 50. (2) Full scan/confirmation mode: Full scans were performed at a resolution of 35,000 (in-source CID: 0 eV), followed by confirmation scans at a resolution of 17,500 using the same stepped NCE values (10, 30, 50).falseSerum and Fecal Metabolomics Analysis in Laboratory-Kenneled PoodlesThis study was conducted to investigate the effects of Lycium barbarum polysaccharides and glycopeptide on the serum and metabolomics of laboratory-kenneled Poodles.2026-03-122026-03-12MTBLS14033