#METABOLOMICS WORKBENCH b72tipam_20251015_000549 DATATRACK_ID:6564 STUDY_ID:ST004299 ANALYSIS_ID:AN007150 PROJECT_ID:PR002715 VERSION 1 CREATED_ON October 17, 2025, 2:58 pm #PROJECT PR:PROJECT_TITLE Mediterranean Quercus acorns as functional foods: Phytochemical profile, PR:PROJECT_TITLE metabolomic insights, and nutritional potential PR:PROJECT_TYPE Untargeted Metabolomics Analysis PR:PROJECT_SUMMARY The present study focuses on the comprehensive phytochemical and metabolomic PR:PROJECT_SUMMARY profiling of acorn flours obtained from eight European Quercus species (Q. ilex PR:PROJECT_SUMMARY subsp. ballota, Q. ilex subsp. ilex, Q. faginea, Q. suber, Q. coccifera, Q. PR:PROJECT_SUMMARY robur, Q. petraea, and Q. pubescens). The dual objective is to support species PR:PROJECT_SUMMARY traceability and to evaluate the nutritional and nutraceutical potential of PR:PROJECT_SUMMARY acorns. PR:INSTITUTE University of Cordoba PR:DEPARTMENT Department of Biochemistry and Molecular Biology PR:LABORATORY AGR-164 PR:LAST_NAME Tienda-Parrilla PR:FIRST_NAME Marta PR:ADDRESS Campus de Rabanales, Edificio C6 Severo Ochoa, Ctra. Madrid, Km 396. 14071 PR:ADDRESS Córdoba, Spain PR:EMAIL b72tipam@uco.es PR:PHONE +34 634925272 #STUDY ST:STUDY_TITLE Mediterranean Quercus acorns as functional foods: Phytochemical profile, ST:STUDY_TITLE metabolomic insights, and nutritional potential ST:STUDY_TYPE Untargeted metabolomics ST:STUDY_SUMMARY The genus Quercus has attracted increasing interest due to its potential for the ST:STUDY_SUMMARY valorization of underutilized natural resources. Most metabolomic studies to ST:STUDY_SUMMARY date have focused on Quercus ilex because of its ecological and economic ST:STUDY_SUMMARY importance in the Mediterranean basin; however, other Mediterranean Quercus ST:STUDY_SUMMARY species may also represent valuable sources of bioactive compounds. In this ST:STUDY_SUMMARY study, we collected acorns from eight Quercus species and analyzed their ST:STUDY_SUMMARY nutritional and phytochemical profiles. Classical biochemical assays based on ST:STUDY_SUMMARY colorimetric reactions were performed, together with near-infrared spectroscopy ST:STUDY_SUMMARY (NIRS) for rapid compositional analysis. We also determined the amino acid ST:STUDY_SUMMARY profiles and conducted untargeted metabolomic profiling using UHPLC-MS/MS after ST:STUDY_SUMMARY metabolite extraction. Our results revealed clear nutritional and metabolomic ST:STUDY_SUMMARY differences associated with leaf habit: evergreen and deciduous species ST:STUDY_SUMMARY displayed distinct metabolic signatures in their acorns. Despite these ST:STUDY_SUMMARY differences, the set of annotated metabolites showed a substantial number of ST:STUDY_SUMMARY compounds common to all species, highlighting Quercus acorns as a rich source of ST:STUDY_SUMMARY nutraceutical molecules, including several flavonoids, hydrolyzable tannins, ST:STUDY_SUMMARY cinnamic acids, coumarins, and terpenoids. ST:INSTITUTE University of Cordoba ST:DEPARTMENT Department of Biochemistry and Molecular Biology ST:LABORATORY AGR-164 ST:LAST_NAME Tienda-Parrilla ST:FIRST_NAME Marta ST:ADDRESS Campus de Rabanales, Edificio C6 Severo Ochoa, Ctra. Madrid, Km 396. 14071 ST:ADDRESS Córdoba, Spain ST:EMAIL b72tipam@uco.es ST:PHONE +34 634925272 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Quercus ilex subsp. ballota, Quercus ilex subsp. ilex, Quercus faginea, Quercus SU:SUBJECT_SPECIES suber, Quercus coccifera, Quercus robur, Quercus petraea, Quercus pubescens SU:TAXONOMY_ID 3110926, 1211604, 501392, 58331, 58335, 38942, 38865, 39471 #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 - Quercus_robur_1_pos Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_1_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_1_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_2_pos Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_2_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_2_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_3_pos Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_3_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240528_3_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_1_pos Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_4_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_4_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_2_pos Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_5_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_5_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_3_pos Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_6_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240528_6_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_1_pos Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_7_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_7_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_2_pos Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_8_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_8_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_3_pos Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_9_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240528_9_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_1_pos Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_10_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_10_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_2_pos Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_11_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_11_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_3_pos Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_12_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240528_12_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_1_pos Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_13_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_13_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_2_pos Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_14_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_14_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_3_pos Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_15_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240528_15_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_1_pos Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_16_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_16_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_2_pos Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_17_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_17_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_3_pos Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_18_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240528_18_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_1_pos Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_19_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_19_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_2_pos Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_20_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_20_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_3_pos Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_21_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240528_21_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_1_pos Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_22_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_22_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_2_pos Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_23_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_23_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_3_pos Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_24_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240528_24_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_1_pos Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_25_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_1_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_25_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_2_pos Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_26_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_2_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_26_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_3_pos Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_27_FS_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_3_pos_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240528_27_TOP5_pos.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_1_neg Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_1_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_1_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_2_neg Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_2_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_2_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_3_neg Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_3_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_robur_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_robur RAW_FILE_NAME(Raw data file)=240531_3_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_1_neg Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_4_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_4_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_2_neg Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_5_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_5_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_3_neg Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_6_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_petraea_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_petraea RAW_FILE_NAME(Raw data file)=240531_6_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_1_neg Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_7_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_7_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_2_neg Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_8_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_8_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_3_neg Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_9_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_pubescens_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_pubescens RAW_FILE_NAME(Raw data file)=240531_9_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_1_neg Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_10_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_10_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_2_neg Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_11_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_11_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_3_neg Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_12_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_faginea_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_faginea RAW_FILE_NAME(Raw data file)=240531_12_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_1_neg Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_13_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_13_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_2_neg Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_14_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_14_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_3_neg Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_15_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_rubra_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_rubra RAW_FILE_NAME(Raw data file)=240531_15_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_1_neg Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_16_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_16_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_2_neg Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_17_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_17_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_3_neg Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_18_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ballota_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ballota RAW_FILE_NAME(Raw data file)=240531_18_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_1_neg Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_19_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_19_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_2_neg Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_20_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_20_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_3_neg Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_21_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_coccifera_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_coccifera RAW_FILE_NAME(Raw data file)=240531_21_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_1_neg Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_22_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_22_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_2_neg Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_23_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_23_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_3_neg Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_24_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_suber_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_suber RAW_FILE_NAME(Raw data file)=240531_24_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_1_neg Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_25_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_1_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_25_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_2_neg Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_26_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_2_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_26_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_3_neg Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_27_FS_neg.mzML SUBJECT_SAMPLE_FACTORS - Quercus_ilex_3_neg_TOP5 Sample source:Fruit_flour | Species:Quercus_ilex RAW_FILE_NAME(Raw data file)=240531_27_TOP5_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_1_neg Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_FS_1_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_2_neg Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_FS_2_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_3_neg Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_FS_3_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_4_neg Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_FS_4_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_1_neg_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_TOP5_1_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_2_neg_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_TOP5_2_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_3_neg_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_TOP5_3_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_4_neg_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240531_QC_MIX_TOP5_4_neg.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_1_pos Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_FS_1_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_2_pos Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_FS_2_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_3_pos Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_FS_3_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_4_pos Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_FS_4_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_1_pos_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_TOP5_1_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_2_pos_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_TOP5_2_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_3_pos_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_TOP5_3_pos.mzML SUBJECT_SAMPLE_FACTORS - QC_MIX_4_pos_TOP5 Sample source:Fruit_flour | Species:QC_MIX RAW_FILE_NAME(Raw data file)=240528_QC_MIX_TOP5_4_pos.mzML #COLLECTION CO:COLLECTION_SUMMARY Acorns from eight European Quercus species (Q. ilex subsp. ballota, Q. ilex CO:COLLECTION_SUMMARY subsp. ilex, Q. faginea, Q. suber, Q. coccifera, Q. robur, Q. petraea, and Q. CO:COLLECTION_SUMMARY pubescens) were kindly provided by the National Centre for Forest Genetic CO:COLLECTION_SUMMARY Resources “El Serranillo” of the Ministry for Ecological Transition and the CO:COLLECTION_SUMMARY Demographic Challenge (Spain). Acorns were harvested at maturity during the CO:COLLECTION_SUMMARY 2023–2024 season. Healthy acorns were surface-sterilized in 10% sodium CO:COLLECTION_SUMMARY hypochlorite for 10 min, rinsed, and stored at 4 °C in darkness. For each CO:COLLECTION_SUMMARY group, ten acorns were measured for weight, volume, length, and width. Acorns CO:COLLECTION_SUMMARY were scarified, seed coats removed, and tissues ground in liquid nitrogen to CO:COLLECTION_SUMMARY obtain flour, later lyophilized and stored in a desiccator at 4 °C until CO:COLLECTION_SUMMARY analysis. Phytochemical assays were performed in triplicate using three CO:COLLECTION_SUMMARY biological replicates, each consisting of a pooled sample of ten acorns. NOTE: CO:COLLECTION_SUMMARY Quercus rubra was included in our experiment as an internal negative control, CO:COLLECTION_SUMMARY since it is a North American species, while all the other species studied are CO:COLLECTION_SUMMARY Mediterranean. It was only used to confirm the geographic origin and distinct CO:COLLECTION_SUMMARY metabolic profile of the other samples. CO:SAMPLE_TYPE Fruit #TREATMENT TR:TREATMENT_SUMMARY No special treatment was employed. Fruits were processed into flour by grinding TR:TREATMENT_SUMMARY in liquid nitrogen. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were extracted from freeze-dried leaf powder. Briefly, a buffer SP:SAMPLEPREP_SUMMARY containing 1200 μL of cold ethanol: water (50:50) was added to 30 mg of flour, SP:SAMPLEPREP_SUMMARY tissue disruption was driven by maceration with pistil, vortexed (10 s) and SP:SAMPLEPREP_SUMMARY sonicated (ultrasonic bath, 40 kHZ for 10 min). After centrifugation (16,000×g, SP:SAMPLEPREP_SUMMARY 4°C, 6 min) the supernatant was vacuum dried at 30 °C (Speedvac, Eppendorf SP:SAMPLEPREP_SUMMARY Vacuum Concentrator Plus/5301, Eppendorf, Leicestershire, UK). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Chromatographic separation was carried out using an Acquity UPLC BEH C18 column CH:CHROMATOGRAPHY_SUMMARY (2.1 x 100 mm, 1.7 µm) (Waters, Manchester, U.K.) maintaining it at 40 °C. The CH:CHROMATOGRAPHY_SUMMARY injection volume was 5 µl and the flow rate was set at 0.5 mL/min. Mobile CH:CHROMATOGRAPHY_SUMMARY phases consisted of 0.1% formic acid in water (Eluent A) and 0.1% formic acid in CH:CHROMATOGRAPHY_SUMMARY methanol (Eluen B). Elution conditions were as follows: 5% B for 1 min, linear CH:CHROMATOGRAPHY_SUMMARY gradient from 5% to 100% in solvent B for 9 min, isocratic at 100% B for 2 min, CH:CHROMATOGRAPHY_SUMMARY and return to initial conditions, 5% B for 3 min. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% methanol; 0.1% formic acid CH:FLOW_GRADIENT 5% B for 1 min, linear gradient from 5% to 100% in solvent B for 9 min, CH:FLOW_GRADIENT isocratic at 100% B for 2 min, and return to initial conditions, 5% B for 3 min. CH:FLOW_RATE 0.5 mL/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS MS detection was performed with the Q Exactive Orbitrap mass spectrometer MS:MS_COMMENTS operating in positive and negative polarities. HESI source parameters in MS:MS_COMMENTS positive mode were spray voltage, 3.5 kV; S-lens RF level, 50; capillary MS:MS_COMMENTS temperature, 320 °C; sheath and auxiliary gas flow, 60 and 25, respectively MS:MS_COMMENTS (arbitrary units); and probe heater temperature, 400 ºC. For negative ion mode, MS:MS_COMMENTS all parameters remained the same except that the spray voltage was set to -3.0 MS:MS_COMMENTS kV. Xcalibur v.4.3 software was used for instrument control and data MS:MS_COMMENTS acquisition. A Full Scan MS method was acquired at a resolution of 70,000 (full MS:MS_COMMENTS width half maximum, FWHM at m/z 200) and a data dependent acquisition MS2 method MS:MS_COMMENTS was acquired at resolution 70,000 and 17,500 (FWHM at m/z 200) for Full Scan and MS:MS_COMMENTS Product Ion Scan, respectively, fragmenting the five most abundant precursor MS:MS_COMMENTS ions per MS scan (Top5). Full Scan MS and data dependent acquisition MS2 methods MS:MS_COMMENTS were acquired in positive and negative modes, and mass range used for both MS:MS_COMMENTS experiments was 70 – 1,050 m/z. Additionally, for continuous quality assurance MS:MS_COMMENTS and to promote confidence in the data, quality control (QC) mix was prepared MS:MS_COMMENTS using equal volumes of all samples and was injected after every six samples MS:MS_COMMENTS during the batch processing along with methanol as a blank run to correct for a MS:MS_COMMENTS drift of the raw signal intensity during the analysis. Moreover, the QC samples MS:MS_COMMENTS were analysed in a data-dependent (dd-MS2/dd-SIM) manner for feature annotation. MS:MS_COMMENTS All acquired data were exported by Xcalibur software to be analysed by the MS:MS_COMMENTS Compound Discoverer v3.2 software (Thermo Fisher Scientific, Bremen, Germany) MS:MS_RESULTS_FILE ST004299_AN007150_Results.txt UNITS:Peak Area Has m/z:Yes Has RT:Yes RT units:Minutes #END