#METABOLOMICS WORKBENCH m183827_20240523_100300 DATATRACK_ID:4859 STUDY_ID:ST003237 ANALYSIS_ID:AN005301 PROJECT_ID:PR002012 VERSION 1 CREATED_ON May 27, 2024, 2:28 pm #PROJECT PR:PROJECT_TITLE Plasm metabolomics of patients diagnosed with Alzheimer's Disease under PR:PROJECT_TITLE Acetylcholinesterase Inhibitors treatment PR:PROJECT_TYPE Untargeted Lipidomics PR:PROJECT_SUMMARY Alzheimer’s Disease (AD) is a fatal neurodegenerative disorder characterized PR:PROJECT_SUMMARY by progressive memory loss, loss of cognitive capacity, mood swings, PR:PROJECT_SUMMARY communication and rationality impairment and eventual loss of independent PR:PROJECT_SUMMARY living. There is still no cure for this disease, but some medicine can delay the PR:PROJECT_SUMMARY progression of symptoms and improve patients’ life quality by acting in the PR:PROJECT_SUMMARY reduction of symptom severity. Acetylcholinesterase inhibitors (AChI) are the PR:PROJECT_SUMMARY main class of drug employed for this. In this context, this research has a goal PR:PROJECT_SUMMARY of evaluating the metabolomic and lipidomic profile of sanguine plasm of PR:PROJECT_SUMMARY patients diagnosed with AD before and after the use of AChI to elucidate PR:PROJECT_SUMMARY metabolic alterations that can assist the diagnosis and contribute to the PR:PROJECT_SUMMARY literature. PR:INSTITUTE State University of Campinas (UNICAMP) PR:DEPARTMENT Institute of Chemistry PR:LABORATORY Laboratory of Bioanalytics and Integated Omics (LaBIOmics) PR:LAST_NAME Marques PR:FIRST_NAME Mariana PR:ADDRESS Rua Josué de Castro, s/n, Campinas, São Paulo, 13083-862, Brazil PR:EMAIL mari31marques@gmail.com PR:PHONE +55 19 35213038 PR:FUNDING_SOURCE Conselho Nacional de Desenvolvimento Científico e Tecnólogico PR:CONTRIBUTORS Alessandra Sussulini, Leda Leme Talib, Wagner Farid Gattaz #STUDY ST:STUDY_TITLE Plasm metabolomics of patients diagnosed with Alzheimer's Disease under ST:STUDY_TITLE Acetylcholinesterase Inhibitors treatment ST:STUDY_TYPE Untargeted Lipidomics Analysis ST:STUDY_SUMMARY Alzheimer’s Disease (AD) is a fatal neurodegenerative disorder characterized ST:STUDY_SUMMARY by progressive memory loss, loss of cognitive capacity, mood swings, ST:STUDY_SUMMARY communication and rationality impairment and eventual loss of independent ST:STUDY_SUMMARY living. There is still no cure for this disease, but some medicine can delay the ST:STUDY_SUMMARY progression of symptoms and improve patients’ life quality by acting in the ST:STUDY_SUMMARY reduction of symptom severity. Acetylcholinesterase inhibitors (AChI) are the ST:STUDY_SUMMARY main class of drug employed for this. In this context, this research has a goal ST:STUDY_SUMMARY of evaluating the metabolomic and lipidomic profile of sanguine plasm of ST:STUDY_SUMMARY patients diagnosed with AD before and after the use of AChI to elucidate ST:STUDY_SUMMARY metabolic alterations that can assist the diagnosis and contribute to the ST:STUDY_SUMMARY literature. The Principal Component Analysis revealed that the posterior ST:STUDY_SUMMARY analysis did not present instrumental biases. As a result, after assessing ST:STUDY_SUMMARY metabolic pathways and putatively identified, statistically significant ST:STUDY_SUMMARY metabolites it was possible to observe some groups of metabolites, such as fatty ST:STUDY_SUMMARY acids and aminoacids, were present both in our data and in literature. Some ST:STUDY_SUMMARY other pathways, such as the biosynthesis of nitrogen and the biosynthesis of ST:STUDY_SUMMARY arginine, are also of interest for research purposes. ST:INSTITUTE University of Campinas ST:DEPARTMENT Institute of Chemistry ST:LABORATORY Laboratory of Bioanalytics and Integated Omics (LaBIOmics) ST:LAST_NAME Marques ST:FIRST_NAME Mariana ST:ADDRESS Rua Josué de Castro, s/n ST:EMAIL mari31marques@gmail.com ST:PHONE +55 19 35213038 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 68-82 SU:GENDER Male and female SU:HUMAN_MEDICATIONS AcetylCholinesterase Inhibitor #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - AD-01 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-01.mzML SUBJECT_SAMPLE_FACTORS - AD-02 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-02.mzML SUBJECT_SAMPLE_FACTORS - AD-03 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-03.mzML SUBJECT_SAMPLE_FACTORS - AD-04 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-04.mzML SUBJECT_SAMPLE_FACTORS - AD-05 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-05.mzML SUBJECT_SAMPLE_FACTORS - AD-06 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-06.mzML SUBJECT_SAMPLE_FACTORS - AD-07 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-07.mzML SUBJECT_SAMPLE_FACTORS - AD-08 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-08.mzML SUBJECT_SAMPLE_FACTORS - AD-09 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-09.mzML SUBJECT_SAMPLE_FACTORS - AD-10 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-10.mzML SUBJECT_SAMPLE_FACTORS - AD-11 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-11.mzML SUBJECT_SAMPLE_FACTORS - AD-12 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-12.mzML SUBJECT_SAMPLE_FACTORS - AD-13 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-13.mzML SUBJECT_SAMPLE_FACTORS - AD-14 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-14.mzML SUBJECT_SAMPLE_FACTORS - AD-15 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-15.mzML SUBJECT_SAMPLE_FACTORS - AD-16 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-16.mzML SUBJECT_SAMPLE_FACTORS - AD-17 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-17.mzML SUBJECT_SAMPLE_FACTORS - AD-18 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-18.mzML SUBJECT_SAMPLE_FACTORS - AD-19 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-19.mzML SUBJECT_SAMPLE_FACTORS - AD-20 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-20.mzML SUBJECT_SAMPLE_FACTORS - AD-21 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-21.mzML SUBJECT_SAMPLE_FACTORS - AD-22 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-22.mzML SUBJECT_SAMPLE_FACTORS - AD-23 AChI dosage:0mg | Sample source:Plasma Group=No treatment; RAW_FILE_NAME(RAW file name)=AD-23.mzML SUBJECT_SAMPLE_FACTORS - AC1-01 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-1.mzML SUBJECT_SAMPLE_FACTORS - AC1-02 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-02.mzML SUBJECT_SAMPLE_FACTORS - AC1-03 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-03.mzML SUBJECT_SAMPLE_FACTORS - AC1-04 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-04.mzML SUBJECT_SAMPLE_FACTORS - AC1-05 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-05.mzML SUBJECT_SAMPLE_FACTORS - AC1-06 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-06.mzML SUBJECT_SAMPLE_FACTORS - AC1-07 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-07.mzML SUBJECT_SAMPLE_FACTORS - AC1-08 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-08.mzML SUBJECT_SAMPLE_FACTORS - AC1-09 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-09.mzML SUBJECT_SAMPLE_FACTORS - AC1-10 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-10.mzML SUBJECT_SAMPLE_FACTORS - AC1-11 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-11.mzML SUBJECT_SAMPLE_FACTORS - AC1-12 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-12.mzML SUBJECT_SAMPLE_FACTORS - AC1-13 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-13.mzML SUBJECT_SAMPLE_FACTORS - AC1-14 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-14.mzML SUBJECT_SAMPLE_FACTORS - AC1-15 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-15.mzML SUBJECT_SAMPLE_FACTORS - AC1-16 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-16.mzML SUBJECT_SAMPLE_FACTORS - AC1-17 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-17.mzML SUBJECT_SAMPLE_FACTORS - AC1-18 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-18.mzML SUBJECT_SAMPLE_FACTORS - AC1-19 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-19.mzML SUBJECT_SAMPLE_FACTORS - AC1-20 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-20.mzML SUBJECT_SAMPLE_FACTORS - AC1-21 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-21.mzML SUBJECT_SAMPLE_FACTORS - AC1-22 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-22.mzML SUBJECT_SAMPLE_FACTORS - AC1-23 AChI dosage:5mg | Sample source:Plasma Group=3 months treatment; RAW_FILE_NAME(RAW file name)=AC1-23.mzML SUBJECT_SAMPLE_FACTORS - AC2-01 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-01.mzML SUBJECT_SAMPLE_FACTORS - AC2-02 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-02.mzML SUBJECT_SAMPLE_FACTORS - AC2-03 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-03.mzML SUBJECT_SAMPLE_FACTORS - AC2-04 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-04.mzML SUBJECT_SAMPLE_FACTORS - AC2-05 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-05.mzML SUBJECT_SAMPLE_FACTORS - AC2-06 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-06.mzML SUBJECT_SAMPLE_FACTORS - AC2-07 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-07.mzML SUBJECT_SAMPLE_FACTORS - AC2-08 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-08.mzML SUBJECT_SAMPLE_FACTORS - AC2-09 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-09.mzML SUBJECT_SAMPLE_FACTORS - AC2-10 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-10.mzML SUBJECT_SAMPLE_FACTORS - AC2-11 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-11.mzML SUBJECT_SAMPLE_FACTORS - AC2-12 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-12.mzML SUBJECT_SAMPLE_FACTORS - AC2-13 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-13.mzML SUBJECT_SAMPLE_FACTORS - AC2-14 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-14.mzML SUBJECT_SAMPLE_FACTORS - AC2-15 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-15.mzML SUBJECT_SAMPLE_FACTORS - AC2-16 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-16.mzML SUBJECT_SAMPLE_FACTORS - AC2-17 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-17.mzML SUBJECT_SAMPLE_FACTORS - AC2-18 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-18.mzML SUBJECT_SAMPLE_FACTORS - AC2-19 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-19.mzML SUBJECT_SAMPLE_FACTORS - AC2-20 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-20.mzML SUBJECT_SAMPLE_FACTORS - AC2-21 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-21.mzML SUBJECT_SAMPLE_FACTORS - AC2-22 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-22.mzML SUBJECT_SAMPLE_FACTORS - AC2-23 AChI dosage:10mg | Sample source:Plasma Group=6 months treatment; RAW_FILE_NAME(RAW file name)=AC2-23.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL01 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL01.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL02 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL02.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL03 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL03.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL04 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL04.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL05 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL05.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL06 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL06.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL07 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL07.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL08 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL08.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL09 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL09.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL10 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL10.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL11 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL11.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL12 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL12.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL13 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL13.mzML SUBJECT_SAMPLE_FACTORS - QCTOTAL14 AChI dosage:Non applicable | Sample source:Plasma(Pooled) Group=Quality Control; RAW_FILE_NAME(RAW file name)=QCTOTAL14.mzML #COLLECTION CO:COLLECTION_SUMMARY The patients for this study were recruited in the ambulatory of Geriatric CO:COLLECTION_SUMMARY research of the Laboratory of Neurosciences in the Faculty of Medical Sciences CO:COLLECTION_SUMMARY of the University of São Paulo (USP). 40 mL of blood were collected in four 10 CO:COLLECTION_SUMMARY mL tubes containing sodium citrate 0.106 mol/L as anticlotting agent. To each CO:COLLECTION_SUMMARY tube 1 mL of ACD-NH was added. The tubes were homogenized and centrifuged for 15 CO:COLLECTION_SUMMARY minutes at 1600 rpm, 20 ºC. Following this, the supernatant was transferred to CO:COLLECTION_SUMMARY a 50 mL Falcon type tube and the pH was adjusted using ACD-NH to 6.5. The plasma CO:COLLECTION_SUMMARY was transferred to 4 polystyrene tubes and centrifuged during 10 minutes at 2400 CO:COLLECTION_SUMMARY rpm, 20ºC. The supernatant was removed by inversion. Plasma aliquots were CO:COLLECTION_SUMMARY storaged at 80 ºC until further analysis. CO:SAMPLE_TYPE Blood (plasma) CO:COLLECTION_FREQUENCY Every 3 months CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS 10 mL containing sodium citrate 0.106 mol/L CO:STORAGE_VIALS polystyrene tubes CO:ADDITIVES Sodium citrate, ACD-NH #TREATMENT TR:TREATMENT_SUMMARY The patients diagnosed with Alzheimer's Disease were subjected to TR:TREATMENT_SUMMARY Acetylcholinesterase Inihibitors treatment with a starting dose of 5mg for 3 TR:TREATMENT_SUMMARY months and 10mg for three more months #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For this study the extraction method used was Simplex. Briefly, 40 µL of plasma SP:SAMPLEPREP_SUMMARY samples were incubated with 300 µL of cold methanol and 1 mL of MTBE (Methyl SP:SAMPLEPREP_SUMMARY tert-butyl ether), followed by the addition of 250 µL of a 0.1% (m/v) ammonium SP:SAMPLEPREP_SUMMARY acetate solution to induce phase separation. Bothe the apolar phase containing SP:SAMPLEPREP_SUMMARY lipids and the polar phase were separated and dried in a vacuum concentrator SP:SAMPLEPREP_SUMMARY (SpeedVac) and stored at -80 °C until further analysis. SP:SAMPLEPREP_PROTOCOL_FILENAME Sample_Preparation.pdf SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD MTBE Simplex (Coman et al, 2016) SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Supelco Titan C18 (1.9 μm, 2.1 x 100 mm) CH:SOLVENT_A 40% acetonitrile/60% water; 10 mM ammonium acetate CH:SOLVENT_B 10% acetonitrile/90% isopropanol;10 mM ammonium acetate CH:FLOW_GRADIENT The elution gradient started with 40% B from minutes 0-2, 50% B from minutes CH:FLOW_GRADIENT 3-6, 70% Bfrom minutes 6.1-8, 100% B from minutes 9-11, and from minutes 12-14 CH:FLOW_GRADIENT the column was stabilized forthe following run CH:FLOW_RATE 0.250 mL/min CH:COLUMN_TEMPERATURE 45 CH:METHODS_FILENAME LC-MS_lipidomics.pdf CH:INJECTION_TEMPERATURE 10 CH:SAMPLE_INJECTION 5 uL CH:ANALYTICAL_TIME 12 minutes CH:CAPILLARY_VOLTAGE 3.5 V #ANALYSIS AN:ANALYSIS_TYPE MS AN:ACQUISITION_DATE 10/31/2023 AN:DATA_FORMAT .raw #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Untargeted lipidomics analysis was performed using an Ultimate 3000 (Thermo) MS:MS_COMMENTS liquid chromatograph coupled to an Orbitrap QExactive (Thermo) mass spectrometer MS:MS_COMMENTS equipped with a heated electrospray ionization (HESI) source operating both in MS:MS_COMMENTS positive (ESI (+)) and negative (ESI (-)) modes using the full-scan mode MS:MS_COMMENTS followed by MS/MS analysis in the data dependent acquisition (DDA) method of the MS:MS_COMMENTS 5 most abundant peaks. The capillary voltage was established at 3.5 V, the MS:MS_COMMENTS drying gas flow at 10 L min-1 and the gas temperature at 300 ºC for the RPLC MS:MS_COMMENTS analysis and 350 ºC for the HILIC analysis. A full scan was performed with a MS:MS_COMMENTS resolution of 70000 and m/z range of 100 to 1500, 1 spectrum was acquired by MS:MS_COMMENTS minute for MS/MS with collision energy of 40 eV. The sampler temperature was set MS:MS_COMMENTS to 10 °C and the column oven temperature was set to 45 °C. MS:CAPILLARY_VOLTAGE 3.5 V MS:COLLISION_ENERGY 40 eV MS:MS_RESULTS_FILE ST003237_AN005301_Results.txt UNITS:Peak Area Has m/z:Yes Has RT:Yes RT units:Minutes #END