MetaboLights

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profiling was performed on a Nexera LC-30A UHPLC system (Shimadzu, Kyoto, Japan). Chromatographic separation was achieved on a Waters ACQUITY UPLC CSH C18 column (Waters Corporation, Milford, MA, USA) maintained at 45 °C with a flow rate of 0.3 mL/min. The mobile phases consisted of acetonitrile/water (6:4, v/v, A) and acetonitrile/isopropanol (1:9, v/v, B). The gradient elution program was set as follows: 0-3.5 min, 40% B; 3.5-13 min, 40-75% B; 13-19 min, 75-99% B; 19-24 min, 40% B.Serum samples were slowly thawed at 4 °C. A 100 μL aliquot of each serum sample was mixed with 400 μL of pre-cooled methanol/acetonitrile (1:1, v/v) and vortexed thoroughly. The mixture was incubated at −20 °C for 30 min to precipitate proteins and then centrifuged at 14,000 × g for 20 min at 4 °C. The supernatant was collected and dried under vacuum. Prior to LC-MS/MS analysis, the dried extract was reconstituted in 100 μL of acetonitrile/water (1:1, v/v), vortexed, and centrifuged at 14,000 × g for 15 min at 4 °C. The resulting supernatant was collected for instrumental analysis. Quality control (QC) samples were prepared by pooling equal volumes of the individual study samples.ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15363Metabolite identification was performed by comparing the experimentally acquired accurate mass-to-charge ratios (m/z) and MS/MS spectra with an in-house metabolite database established using available authentic standards. Metabolite annotations were assigned based on high-resolution accurate mass matching with a mass error of <10 ppm together with MS/MS spectral matching.Data were processed using LipidSearch software (Thermo Fisher Scientific) for peak alignment, extraction and identification. Main parameters were set as follows: precursor tolerance = 5 ppm, product tolerance = 5 ppm, and product ion threshold = 5%. Quantitative analysis was performed using an internal standard approach, and the absolute concentration of each lipid species was calculated from the response ratio (peak area ratio) of analyte to the internal standard and its known concentration.Data were processed using LipidSearch software (Thermo Fisher Scientific) for peak alignment, extraction and identification. Main parameters were set as follows: precursor tolerance = 5 ppm, product tolerance = 5 ppm, and product ion threshold = 5%. Quantitative analysis was performed using an internal standard approach, and the absolute concentration of each lipid species was calculated from the response ratio (peak area ratio) of analyte to the internal standard and its known concentration.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - positive - reverse-phaseLiquid Chromatography MS - negative - hilicMass spectrometric analysis was performed using a Q-Exactive Plus mass spectrometer (Thermo Fisher Scientific, USA) equipped with an electrospray ionization (ESI) source. Data were acquired separately in positive and negative ionization modes. The ESI source parameters were set as follows: heater temperature, 300 °C; capillary temperature, 350 °C; spray voltage, 3.0 kV; sheath gas flow rate, 45 arbitrary units (arb); auxiliary gas flow rate, 15 arb; sweep gas flow rate, 1 arb; and S-Lens RF level, 50%. The MS1 scan range was m/z 200–1800. Following each full MS1 scan, 10 MS2 fragment spectra were acquired using higher-energy collisional dissociation (HCD). The resolutions of MS1 and MS2 were set to 70,000 and 17,500 at m/z 200, respectively.Mass spectrometry was performed on a Q Exactive™ Plus mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA), with data acquired in positive (ESI+) and negative (ESI-) ionization modes. The MS scan range was set to 200-1800 m/z, and ten MS² (HCD) fragment spectra were acquired following each full MS¹ scan. Resolutions for MS¹ and MS² were 70,000 and 17,500, respectively.Untargeted metabolomic analysis was performed using an Agilent 1290 Infinity LC UHPLC system (Agilent Technologies, USA) with hydrophilic interaction liquid chromatography (HILIC) separation on an ACQUITY UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm; Waters Corporation). The column temperature was maintained at 25 °C, the flow rate was 0.5 mL/min, and the injection volume was 2 μL. Mobile phase A consisted of water containing 25 mM ammonium acetate and 25 mM ammonium hydroxide, while mobile phase B was acetonitrile. The gradient was as follows: 0–0.5 min, 95% B; 0.5–5.0 min, 95–65% B; 5.0–5.5 min, 65–40% B; 5.5–6.0 min, 40–95% B; and 6.0–8.0 min, 95% B. Samples were maintained in the autosampler at 4 °C and analyzed in randomized order, with QC samples interspersed throughout the analytical sequence to monitor system stability.walnut protein hydrolysate alleviates neuroinflammation-associated memory impairment in naturally aged mice through Akkermansia-lysophosphatidylcholine metabolism-NLRP3 inflammasome pathway.yao tangKunming University of Science and Technologyblood serummass spectrometry assayAn appropriate amount of serum sample was mixed with 200 μL of water, followed by the addition of 800 μL MTBE and 240 μL of pre-cooled methanol. The mixture was vortexed, sonicated in a low-temperature water bath for 20 min, and allowed to stand at room temperature for 30 min. After centrifugation at 14,000 × g for 15 min at 10 °C, the upper organic phase was collected and dried under nitrogen. The dried extract was reconstituted in 200 μL of 90% isopropanol/acetonitrile solution. Subsequently, 90 μL of the reconstituted solution was centrifuged at 14,000 × g for 15 min at 10 °C, and the supernatant was used for LC-MS/MS analysis.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS15363yao tang. Kunming University of Science and Technology. tangyao3874@163.com.Raw LC-MS/MS data were processed using LipidSearch software (Thermo Fisher Scientific). Lipid features and internal standard lipid features were subjected to peak alignment, peak identification, and peak extraction to generate a data matrix containing mass-to-charge ratio (m/z), retention time, and peak intensity. Peak intensities were batch-normalized to the total spectral intensity to facilitate comparison among samples.TreatmentGenotypeAgetangyao3874@163.comSerum samples were collected from mice included in the natural ageing, dietary intervention, fecal microbiota transplantation (FMT), and LPC 18:2 supplementation experiments. For the untargeted metabolomics analysis, serum samples were obtained from 2-month-old, 10-month-old, and 18-month-old C57BL/6J mice, including 18-month-old control mice and mice treated with walnut protein (WP) or walnut protein hydrolysate (WPH). For the lipidomics analysis, serum samples were obtained from 18-month-old C57BL/6J mice in the control, FMT, and LPC 18:2 supplementation groups, as well as from 4-month-old APP/PS1 mice in the control, FMT, and LPC 18:2 supplementation groups. The collected serum samples were subsequently subjected to untargeted metabolomic or lipidomic analysis according to the corresponding experimental design.Mass spectrometric analysis was performed using an AB Sciex TripleTOF 6600 mass spectrometer equipped with an electrospray ionization (ESI) source. Data were acquired in both positive and negative ionization modes. The ESI source parameters were set as follows: ion source gas 1 (Gas1), 60; ion source gas 2 (Gas2), 60; curtain gas (CUR), 30 psi; source temperature, 600 °C; and ion spray voltage floating (ISVF), +5500 V in positive mode and −5500 V in negative mode. The MS1 mass range was m/z 60–1000, with an accumulation time of 0.20 s/spectrum. MS/MS spectra were acquired over m/z 25–1000 with an accumulation time of 0.05 s/spectrum using information-dependent acquisition (IDA). The declustering potential was set to ±60 V, and the collision energy was 35 ± 15 eV. Isotope ions within 4 Da were dynamically excluded, and up to 10 fragment spectra were acquired per scan.Total lipids from serum were extracted using the methyl tert-butyl ether (MTBE) method. Approximately 30 μL of serum was mixed with 200 μL of water and vortexed for 5 s. Subsequently, 240 μL of pre-cooled methanol (MeOH) was added and vortexed for 30 s. Next, 800 μL of MTBE was added, and the mixture was sonicated at 4 °C for 20 min, followed by standing at room temperature for 30 min. The mixture was centrifuged at 14,000 × g for 15 min at 10 °C, and the upper organic phase was collected and evaporated to dryness under a gentle nitrogen (N2) stream. The dried lipid residue was reconstituted in 200 μL of 90% isopropanol/acetonitrile (IPA/ACN), vortexed thoroughly and centrifuged again at 14,000 × g for 15 min at 10 °C. The supernatant was collected for LC-MS/MS analysis.Lipid species were identified using LipidSearch software based on accurate mass (m/z) and MS/MS fragment ion matching.MetabolomicsRaw LC-MS/MS data were converted to an open mass spectrometry data format using ProteoWizard MSConvert and subsequently processed using XCMS for peak detection, retention time correction, peak alignment, and peak area extraction. For peak picking, the centWave algorithm was used with an m/z tolerance of 10 ppm, peak width of 10–60 s, and prefilter = c(10, 100). For peak grouping, the parameters were set to bw = 5, mzwid = 0.025, and minfrac = 0.5. Features with more than 50% missing values were removed, remaining missing values were imputed using the K-nearest neighbors (KNN) method, and features with a relative standard deviation (RSD) greater than 50% were excluded prior to subsequent statistical analysis.AB Sciex TripleTOF 6600 mass spectrometerpooled quality control sampleMetabolomicsMus musculusuntargeted analysisAgilent 1290 Infinity LC UHPLC systemLipidomicsLipid MetabolismNexera LC-30A UHPLC system (Shimadzu, Kyoto, Japan)experimental sampleMouseblood serumAgingQ Exactive Plus mass spectrometer (Thermo Fisher Scientific)AB Sciex TripleTOF 6600 mass spectrometerpooled quality control sampleMetabolomicsMus musculusuntargeted analysisAgilent 1290 Infinity LC UHPLC systemLipidomicsLipid MetabolismNexera LC-30A UHPLC system (Shimadzu, Kyoto, Japan)experimental sampleMouseblood serumAgingQ Exactive Plus mass spectrometer (Thermo Fisher Scientific)Mass spectrometric analysis was performed using a Q Exactive Plus mass spectrometer (Thermo Fisher Scientific, USA) equipped with an electrospray ionization (ESI) source. Data were acquired separately in positive (ESI+) and negative (ESI-) ionization modes. The ESI source parameters were set as follows: heater temperature, 300 degrees C; capillary temperature, 350 degrees C; spray voltage, 3.0 kV; sheath gas flow rate, 45 arbitrary units (arb); auxiliary gas flow rate, 15 arb; sweep gas flow rate, 1 arb; and S-Lens RF level, 50%. The MS1 scan range was m/z 200-1800. Following each full MS1 scan, 10 MS2 fragment spectra were acquired using higher-energy collisional dissociation (HCD). The resolutions of MS1 and MS2 were set to 70,000 and 17,500 at m/z 200, respectively.TG(58:11)+NH4LPC(17:0)+HLPC(22:6)+HPC(42:11)+HLPC(18:3)+HPC(36:7)+HLPC(24:1)+HPC(46:6)+HLPC(20:0)+HTG(50:6)+NH4PC(34:4)+HPC(37:2)+HTG(49:5)+NaDG(36:0)+NH4LPC(18:0)+NaPC(44:6)+HDG(21:0)+NaDG(41:0)+NaPI(40:3)+NaPC(37:2)+NaPC(38:8)+HPC(34:2)+NaPC(31:1)+HPC(40:8)+HPC(34:1)+HPC(38:0)+HPC(35:4)+HPE(46:5)+NaDG(36:4)+HPC(37:5)+HLPC(20:3)+HPC(36:1)+HLPC(22:4)+HLPE(20:3)+NaLPC(14:0)+HPC(38:6)+NaMG(20:2)+HCL(80:10)+HPC(38:7)+HPC(35:5)+HMG(22:2)+NH4PG(38:6)+NH4PC(31:0)+HPC(42:10)+HLPC(15:0)+HPC(34:6)+HTG(58:9)+NH4PC(44:12)+HDG(42:6)+NH4LPE(20:1)+NaTG(54:12)+NaLPC(24:0)+HLPC(20:1)+HDG(34:2)+NH4PG(36:1)+HPC(38:2)+HLPC(22:0)+NaPA(39:0)+NH4PC(34:3)+HPE(22:1)+HPC(42:7)+HTG(59:7)+NH4PC(40:6)+HDG(38:7)+HPI(40:4)+NH4PC(39:1)+HLPC(22:2)+HPC(38:9)+HPC(40:6)+NaPC(36:3)+HPC(20:0)+HPI(40:3)+NH4DG(40:6)+NaLPE(22:6)+HPC(35:3)+HPC(40:9)+HLPI(18:1)+NaPC(37:6)+HLPC(20:4)+HPC(33:2)+HLPC(16:0)+NaLPC(22:5)+HLPC(18:2)+HDG(34:3)+HPC(36:6)+HLPC(22:6)+NaLPE(20:4)+NaPC(39:6)+HPC(42:9)+HPC(34:5)+HPC(32:0)+HPI(36:3)+NaPC(33:3)+HPC(44:11)+HLPC(22:0)+HLPC(18:1)+HLPC(20:0)+NaPC(36:5)+HPC(36:4)+NaDG(48:0)+NH4PC(38:3)+HPC(46:7)+HPI(38:3)+NH4PI(40:5)+NH4PC(40:7)+HPC(42:6)+HPC(32:3)+HPC(37:4)+HPC(33:0)+HTG(49:3)+NH4TG(56:6)+NH4TG(56:3)+NH4PC(36:8)+HMG(18:1)+NaPC(42:3)+HPE(34:1)+NaDG(36:4)+NaTG(53:7)+NH4PC(40:2)+HPA(33:0)+NH4LPC(20:5)+HfalseWalnut protein hydrolysate alleviates neuroinflammation and memory impairment in naturally aged mice via a gut microbiota-metabolite-neuroinflammation axisThis study investigated age-associated alterations in the serum metabolome and the metabolic effects of walnut protein hydrolysate (WPH) supplementation in naturally aged mice. Male C57BL/6J mice at different ages (2-month, 10-month and 18-month) were used to characterize metabolic changes associated with natural ageing. Eighteen-month-old mice were further administered walnut protein (WP) or walnut protein hydrolysate (WPH) to evaluate their effects on ageing-associated metabolic alterations. Serum samples were collected and subjected to untargeted metabolomic analysis to characterize global metabolic profiles and identify metabolites associated with ageing and WPH intervention. Differential metabolite analysis and metabolic pathway enrichment analysis were performed to characterize metabolic pathways associated with ageing and WPH treatment, with particular attention to lipid metabolism and phosphatidylcholine/lysophosphatidylcholine metabolism. The identified metabolic alterations were further investigated in relation to gut microbiota remodeling and memory-related phenotypes. Additional validation experiments involving fecal microbiota transplantation and LPC 18:2 supplementation were performed to investigate the potential role of gut microbiota-associated LPC 18:2 metabolism in the memory-improving effects of WPH. This dataset provides serum metabolomic data supporting the investigation of the gut microbiota–metabolite–brain axis underlying the effects of WPH on ageing-associated memory impairment.2026-09-042026-08-16MTBLS15363HMDB0010379HMDB0240262HMDB0010383HMDB0012108HMDB0011149HMDB0061701HMDB0061700HMDB0010388HMDB0010390HMDB0010391HMDB0010392HMDB0010394HMDB0061699HMDB0010397HMDB0010398HMDB0010399HMDB0010401HMDB0010404HMDB0010405HMDB0010406HMDB0010403HMDB0010381HMDB0008502HMDB0008744HMDB0013422HMDB0013443HMDB0008738HMDB0008292HMDB0034202HMDB0001124HMDB0245844HMDB0002520HMDB0304597HMDB0003073HMDB0005783HMDB0011152HMDB0011394HMDB0060631HMDB0251497HMDB0010395HMDB0011507HMDB0007859HMDB0061695HMDB0243892HMDB0001563HMDB0003099HMDB0007869HMDB0114884HMDB0240606HMDB0013426HMDB0008102HMDB0007982HMDB0011503HMDB0011130HMDB0007861HMDB0008039HMDB0008994HMDB0008031HMDB0008038HMDB0002327HMDB0002107HMDB0060497HMDB0007865HMDB0010565HMDB0244815HMDB0244212HMDB0244228HMDB0247218HMDB0244269HMDB0244347HMDB0304547HMDB0003871HMDB0006294HMDB0000374HMDB0034517HMDB0010214HMDB0002302HMDB0250341HMDB0011747HMDB0062766HMDB0304072HMDB0000402HMDB0001864HMDB0000354HMDB0005862HMDB0255446HMDB0000225HMDB0000397HMDB0014706HMDB0004811HMDB0240715HMDB0302338HMDB0013676HMDB0029673HMDB0000905HMDB0001245HMDB0001202HMDB0001409HMDB0004326HMDB0060727HMDB0255802HMDB0005998HMDB0247339HMDB0245790HMDB0245953HMDB0253211HMDB0304121HMDB0012710HMDB0000425HMDB0000732HMDB0061656HMDB0001368HMDB0000019HMDB0245967HMDB0000026HMDB0000423HMDB0001866HMDB0034004HMDB0034069HMDB0246682HMDB0012454HMDB0002094HMDB0246399HMDB0304180HMDB0245269HMDB0060390HMDB0246466HMDB0248042HMDB0240465HMDB0002916HMDB0041950HMDB0060272HMDB0000543HMDB0000017HMDB0246588HMDB0240711HMDB0004816HMDB0245221HMDB0246634HMDB0030090HMDB0041905HMDB0003355HMDB0060396HMDB0060542HMDB0030637HMDB0002339HMDB0011725HMDB0036620HMDB0255448HMDB0001983HMDB0005082HMDB0041812HMDB0247037HMDB0004081HMDB0062800HMDB0001316HMDB0000876HMDB0304330HMDB0242135HMDB0240687HMDB0011107HMDB0001991HMDB0000501HMDB0002275HMDB0247297HMDB0006218HMDB0004704HMDB0014797HMDB0015324HMDB0247898HMDB0243752HMDB0035687HMDB0001494HMDB0011756HMDB0014925HMDB0000538HMDB0248010HMDB0000448HMDB0036561HMDB0251509HMDB0013209HMDB0060561HMDB0000037HMDB0260413HMDB0037324HMDB0006254HMDB0000462HMDB0035219HMDB0039353HMDB0000975HMDB0248258HMDB0248260HMDB0248277HMDB0240707HMDB0015440HMDB0243588HMDB0033896HMDB0000053HMDB0004484HMDB0248432HMDB0303733HMDB0132454HMDB0002212HMDB0248571HMDB0304777HMDB0028707HMDB0028713HMDB0028717HMDB0240267HMDB0028731HMDB0028739HMDB0028741HMDB0028743HMDB0028744HMDB0028748HMDB0028759HMDB0028764HMDB0001894HMDB0006483HMDB0303169HMDB0005006HMDB0041830HMDB0036562HMDB0000784HMDB0041832HMDB0041833HMDB0248960HMDB0248961HMDB0244688HMDB0304270HMDB0242697HMDB0014702HMDB0038419HMDB0003409HMDB0000056HMDB0015465HMDB0015260HMDB0001008HMDB0030833HMDB0002338HMDB0000817HMDB0002114HMDB0032133HMDB0249276HMDB0249277HMDB0035074HMDB0248832HMDB0249353HMDB0060928HMDB0014382HMDB0033826HMDB0003501HMDB0013680HMDB0000900HMDB0249566HMDB0014934HMDB0249587HMDB0249590HMDB0000511HMDB0014704HMDB0242751HMDB0249633HMDB0014959HMDB0002780HMDB0249707HMDB0014971HMDB0015422HMDB0014675HMDB0002385HMDB0000055HMDB0014599HMDB0249817HMDB0249836HMDB0249899HMDB0033938HMDB0014436HMDB0014589HMDB0002394HMDB0000653HMDB0012199HMDB0036619HMDB0014513HMDB0247784HMDB0004078HMDB0250268HMDB0250276HMDB0003229HMDB0000072HMDB0006331HMDB0000749HMDB0000094HMDB0041857HMDB0034844HMDB0014904HMDB0250329HMDB0015466HMDB0011680HMDB0001261HMDB0250451HMDB0031457HMDB0250485HMDB0002269HMDB0031340HMDB0030093HMDB0036423HMDB0028773HMDB0000731HMDB0248553HMDB0000089HMDB0000095HMDB0000143HMDB0003391HMDB0001151HMDB0000539HMDB0029942HMDB0001851HMDB0001321HMDB0000660HMDB0304311HMDB0000639HMDB0062783HMDB0000663HMDB0303730HMDB0000131HMDB0006355HMDB0003402HMDB0000169HMDB0304312HMDB0001498HMDB0000845HMDB0003374HMDB0003411HMDB0250793HMDB0247051HMDB0001548HMDB0244230HMDB0003418HMDB0004136HMDB0000098HMDB0240698HMDB0033991HMDB0015350HMDB0014395HMDB0060984HMDB0255779HMDB0250946HMDB0033801HMDB0000626HMDB0250725HMDB0001342HMDB0251222HMDB0014884HMDB0037479HMDB0003581HMDB0251200HMDB0251202HMDB0014628HMDB0094660HMDB0251254HMDB0039721HMDB0000038HMDB0303757HMDB0251309HMDB0030847HMDB0240498HMDB0015203HMDB0000079HMDB0001882HMDB0002050HMDB0029676HMDB0006692HMDB0251505HMDB0060475HMDB0014979HMDB0000933HMDB0251621HMDB0011632HMDB0251686HMDB0251690HMDB0033022HMDB0041397HMDB0029861HMDB0005056HMDB0003153HMDB0251854HMDB0015014HMDB0243678HMDB0003045HMDB0005811HMDB0005810HMDB0036785HMDB0002068HMDB0002994HMDB0004483HMDB0001425HMDB0010325HMDB0033788HMDB0303556HMDB0252085HMDB0015210HMDB0000961HMDB0252185HMDB0252201HMDB0014938HMDB0252265HMDB0252267HMDB0003075HMDB0015326HMDB0252322HMDB0252328HMDB0252332HMDB0252340HMDB0252414HMDB0014642HMDB0252429HMDB0005808HMDB0015399HMDB0252457HMDB0242559HMDB0014635HMDB0252473HMDB0001076HMDB0002741HMDB0242599HMDB0000134HMDB0034702HMDB0001933HMDB0252558HMDB0003464HMDB0005826HMDB0000565HMDB0005807HMDB0252605HMDB0034095HMDB0001049HMDB0011737HMDB0034931HMDB0035988HMDB0252641HMDB0030700HMDB0252648HMDB0015178HMDB0014584HMDB0252679HMDB0003217HMDB0059999HMDB0004486HMDB0003559HMDB0303447HMDB0303451HMDB00357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