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Mouse serum samples from the NFD cohort , were thawed on ice prior to lipid extraction. For extraction, 400 μL of 80% isopropanol (IPA) was added to 50 μL of each sample. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , Mouse serum samples from the CDAA cohort , were thawed on ice prior to lipid extraction. For extraction, 400 μL of 80% isopropanol (IPA) was added to 50 μL of each sample. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , Mouse serum samples from the FPC cohort , were thawed on ice prior to lipid extraction. For extraction, 400 μL of 80% isopropanol (IPA) was added to 50 μL of each sample. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , Mouse Liver tissue samples from the NFD cohort , were thawed on ice prior to lipid extraction. Tissues were weighed and homogenized at a ratio of 1 mg tissue per 3 μL solvent using a Biomasher drill. The homogenates were centrifuged for 5 min at 18,341 × g and 4 °C and the resulting 200uL supernatants were collected for extraction. For extraction, 200 μL of Water and methyl tert-butyl ether (MTBE) 600uL was added to 200 μL of each homogenates. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. For tissue lipidomics, 20 μL of tissue homogenate was mixed with 10 μL of internal standard (IS) solution, followed by the addition of 200 μL of 80% isopropanol (IPA). The mixtures were vortexed and centrifuged, and 180 μL of the supernatant was transferred to a new tube. Subsequently, liquid–liquid extraction was performed by adding 200 μL of distilled water and 600 μL of MTBE (water:IPA:MTBE = 1:1:3). Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , Mouse Liver tissue samples from the CDAA cohort , were thawed on ice prior to lipid extraction. Tissues were weighed and homogenized at a ratio of 1 mg tissue per 3 μL solvent using a Biomasher drill. The homogenates were centrifuged for 5 min at 18,341 × g and 4 °C and the resulting 200uL supernatants were collected for extraction. For extraction, 200 μL of Water and methyl tert-butyl ether (MTBE) 600uL was added to 200 μL of each homogenates. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. For tissue lipidomics, 20 μL of tissue homogenate was mixed with 10 μL of internal standard (IS) solution, followed by the addition of 200 μL of 80% isopropanol (IPA). The mixtures were vortexed and centrifuged, and 180 μL of the supernatant was transferred to a new tube. Subsequently, liquid–liquid extraction was performed by adding 200 μL of distilled water and 600 μL of MTBE (water:IPA:MTBE = 1:1:3). Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , Mouse Liver tissue samples from the FPC cohort , were thawed on ice prior to lipid extraction. Tissues were weighed and homogenized at a ratio of 1 mg tissue per 3 μL solvent using a Biomasher drill. The homogenates were centrifuged for 5 min at 18,341 × g and 4 °C and the resulting 200uL supernatants were collected for extraction. For extraction, 200 μL of Water and methyl tert-butyl ether (MTBE) 600uL was added to 200 μL of each homogenates. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. For tissue lipidomics, 20 μL of tissue homogenate was mixed with 10 μL of internal standard (IS) solution, followed by the addition of 200 μL of 80% isopropanol (IPA). The mixtures were vortexed and centrifuged, and 180 μL of the supernatant was transferred to a new tube. Subsequently, liquid–liquid extraction was performed by adding 200 μL of distilled water and 600 μL of MTBE (water:IPA:MTBE = 1:1:3). Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis. , For each cohort, extracted samples were pooled prior to lipid extraction. For extraction, 400 μL of 80% isopropanol (IPA) was added to 50 μL of each sample. The mixtures were vortexed and centrifuged for 5 min at 18,341 × g and 4 °C. Subsequently, 200 μL of the supernatant was transferred into two Eppendorf SafeLock tubes and dried under N2 gas. The dried pellets were reconstituted with 100 μL of 95% acetonitrile (ACN) containing 10 mM ammonium acetate (AmAc) for HILIC-MRM analysis.

Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 2 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 3 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 4 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 5 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 6 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 7 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 8 μL. , Chromatographic separation was achieved on an ACQUITY Premier BEH Amide column (1.7 μm, 100 × 2.1 mm I.D., Waters, Milford, MA, USA) using mobile phases A (ACN:Water = 95:5) and B (ACN:Water = 50:50), both containing 10 mM AmAc. The injector needle was washed with ACN:Water (95:5) after each injection, and the injection volume was set to 2 μL. The gradient elution program was as follows: the system was initially maintained at 0.1% B for 2 min, followed by an increase to 80% B over 3 min. The gradient was returned to the initial condition (0.1% B) within 0.1 min and equilibrated for 3 min at the starting condition. The flow rate was set to 300 μL/min, the column temperature was maintained at 40 °C, and the sample injection volume was 9 μL.

Optimized ion source parameters included a voltage of 5500 V in positive mode and −4500 V in negative mode, an ion source temperature of 400 °C, and GS1 and GS2 settings of 30 and 35 psi, respectively. Curtain gas and collision-activated dissociation gas pressures were maintained at 20 and 6 psi. A scheduled MRM (sMRM) algorithm with a retention time (tR) window of 1 min was applied.