#METABOLOMICS WORKBENCH b72tipam_20231218_085820 DATATRACK_ID:4535 STUDY_ID:ST003016 ANALYSIS_ID:AN004949 PROJECT_ID:PR001877 VERSION 1 CREATED_ON December 19, 2023, 1:43 am #PROJECT PR:PROJECT_TITLE Metabolomics analysis of Cormus domestica (L.) fruits and the valorisation of an PR:PROJECT_TITLE ethnobotanical heritage of culinary and medicinal uses in Italy PR:PROJECT_TYPE LC-MS analysis PR:PROJECT_SUMMARY Cormus domestica (L.) is a monophyletic wild fruit tree belonging to the PR:PROJECT_SUMMARY Rosaceae family, with well-documented use in the Mediterranean region. PR:PROJECT_SUMMARY Traditionally, these fruits are harvested and stored for at least 2 weeks before PR:PROJECT_SUMMARY consumption. During this period, the fruit reaches its well-known and peculiar PR:PROJECT_SUMMARY organoleptic and texture characteristics. However, the spread of more PR:PROJECT_SUMMARY profitable fruit tree species, resulted in its progressive erosion. In this work PR:PROJECT_SUMMARY we performed proteomic and metabolomic fruit analyses at three times after PR:PROJECT_SUMMARY harvesting to provide data on its chemical composition and nutritional and PR:PROJECT_SUMMARY nutraceutical properties. PR:INSTITUTE Università degli Studi del Sannio (Benevento, Italy) PR:DEPARTMENT University of Sannio, Department of Science and Technology, via de Sanctis, PR:DEPARTMENT Benevento 82100 Italy AND University of Córdoba, Department of Biochemistry PR:DEPARTMENT and Molecular Biology, Spain PR:LABORATORY Integrated Laboratory of Animal and Plant Biology for Environment and Health PR:LABORATORY (Italy) AND Agroforestry and Plant Biochemistry, Proteomics and Systems Biology PR:LABORATORY (Spain) PR:LAST_NAME Tienda PR:FIRST_NAME Marta PR:ADDRESS Pz/ Escultor Ruiz Olmos 2, 5-3 PR:EMAIL b72tipam@uco.es PR:PHONE 634925272 PR:FUNDING_SOURCE This research was funded by Progetto DICOVALE (CUP B24I19000440009), Programma PR:FUNDING_SOURCE di Sviluppo Rurale 2014-2020. SOTTOMISURA 10.2.1 “Conservazione delle risorse PR:FUNDING_SOURCE generiche autoctone a tutela della biodiversità – RGV Risorse PR:FUNDING_SOURCE Genetiche Vegetali” #STUDY ST:STUDY_TITLE Metabolomics analysis of Cormus domestica (L.) fruits and the valorisation of an ST:STUDY_TITLE ethnobotanical heritage of culinary and medicinal uses in Italy ST:STUDY_TYPE Untargeted MS-based metabolomics ST:STUDY_SUMMARY Cormus domestica (L.) is a monophyletic wild fruit tree belonging to the ST:STUDY_SUMMARY Rosaceae family, with well-documented use in the Mediterranean region. ST:STUDY_SUMMARY Traditionally, these fruits are harvested and stored for at least 2 weeks before ST:STUDY_SUMMARY consumption. During this period, the fruit reaches its well-known and peculiar ST:STUDY_SUMMARY organoleptic and texture characteristics. However, the spread of more ST:STUDY_SUMMARY profitable fruit tree species, resulted in its progressive erosion. In this work ST:STUDY_SUMMARY we performed proteomic and metabolomic fruit analyses at three times after ST:STUDY_SUMMARY harvesting to provide data on its chemical composition and nutritional and ST:STUDY_SUMMARY nutraceutical properties. ST:INSTITUTE Università degli Studi del Sannio (Benevento, Italy) and University of Córdoba ST:INSTITUTE (Spain) ST:DEPARTMENT University of Sannio, Department of Science and Technology, via de Sanctis, ST:DEPARTMENT Benevento 82100 Italy AND University of Córdoba, Department of Biochemistry ST:DEPARTMENT and Molecular Biology, Spain ST:LABORATORY Integrated Laboratory of Animal and Plant Biology for Environment and Health ST:LABORATORY (Italy) AND Agroforestry and Plant Biochemistry, Proteomics and Systems Biology ST:LABORATORY (Spain) ST:LAST_NAME Tienda ST:FIRST_NAME Marta ST:ADDRESS Campus de Rabanales, Córdoba, Córdoba, 14014, Spain ST:EMAIL b72tipam@uco.es ST:PHONE 634925272 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Cormus domestica (L.) SU:TAXONOMY_ID 36606 #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 - 1_FS Factor:T1 RAW_FILE_NAME=220923_1_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 2_FS Factor:T1 RAW_FILE_NAME=220923_2_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 3_FS Factor:T1 RAW_FILE_NAME=220923_3_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 4_FS Factor:T2 RAW_FILE_NAME=220923_4_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 5_FS Factor:T2 RAW_FILE_NAME=220923_5_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 6_FS Factor:T2 RAW_FILE_NAME=220923_6_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 7_FS Factor:T3 RAW_FILE_NAME=220923_7_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 8_FS Factor:T3 RAW_FILE_NAME=220923_8_FS_pos.mzXML SUBJECT_SAMPLE_FACTORS - 9_FS Factor:T3 RAW_FILE_NAME=220923_9_FS_pos.mzXML #COLLECTION CO:COLLECTION_SUMMARY The fruits of Cormus domestica (L.) were collected in Campania, Italy in CO:COLLECTION_SUMMARY September 2021 from three different trees located in the same area with slightly CO:COLLECTION_SUMMARY alkaline, sandy and well drained soils (Figure 5). The climate in this area is CO:COLLECTION_SUMMARY typically Mediterranean, with average minimum winter temperatures of 8°C and CO:COLLECTION_SUMMARY 24°C in summer and average annual rainfall of around 900 mm. The harvested CO:COLLECTION_SUMMARY fruits (T0) were transported to the laboratory where the traditional CO:COLLECTION_SUMMARY post-harvest ripening procedure for these fruits was applied, which involves CO:COLLECTION_SUMMARY staying in a ventilated, cool and dry place in the dark for a period of about 2 CO:COLLECTION_SUMMARY weeks (T1). Ripening continued for a further 2 weeks until the fruit had CO:COLLECTION_SUMMARY completely turned brown (T2). CO:SAMPLE_TYPE Plant CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY The harvested fruits (T1) were transported to the laboratory where the TR:TREATMENT_SUMMARY traditional post-harvest ripening procedure for these fruits was applied, which TR:TREATMENT_SUMMARY involves staying in a ventilated, cool and dry place in the dark for a period of TR:TREATMENT_SUMMARY about 2 weeks (T2). Ripening continued for a further 2 weeks until the fruit had TR:TREATMENT_SUMMARY completely turned brown (T3). TR:PLANT_GROWTH_LOCATION Campania, Italy in September 2021 from three different trees located in the same TR:PLANT_GROWTH_LOCATION area with slightly alkaline, sandy and well drained soils TR:PLANT_METAB_QUENCH_METHOD Liquid N2 TR:PLANT_STORAGE Freeze-dried #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For metabolite extraction, three replicates were taken for each ripening stage SP:SAMPLEPREP_SUMMARY considered (T1-T2-T3). Fruits were washed in deionised water, freeze-dried and SP:SAMPLEPREP_SUMMARY pulverised in liquid nitrogen. Metabolites were then extracted from 30 SP:SAMPLEPREP_SUMMARY milligrams of sample using 600 microlitres of cold ethanol:water (4:1) SP:SAMPLEPREP_SUMMARY extraction solution while stirring for 10 s. The mixture was sonicated (40 kHZ SP:SAMPLEPREP_SUMMARY for 10 min) and centrifuged (20,000 g, 4°C, 4 min), the supernatant thus SP:SAMPLEPREP_SUMMARY obtained was transferred to a new tube and the extraction was repeated on the SP:SAMPLEPREP_SUMMARY pellet with a further 600 microlitres of ethanol:water (4:1). SP:PROCESSING_METHOD Ethanol:Water SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Ethanol:Water SP:EXTRACT_CLEANUP Centrifugation SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION Methanol SP:SAMPLE_DERIVATIZATION NO SP:SAMPLE_SPIKING NO #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Chromatographic separations were performed using an Acquity UPLC BEH C18 column CH:CHROMATOGRAPHY_SUMMARY (2.1 x 100 mm, 1.7 µm) (Waters). The column was maintained at 40 °C and eluted CH:CHROMATOGRAPHY_SUMMARY under the following conditions: 5% B for 1 min, linear gradient from 5% to 100% CH:CHROMATOGRAPHY_SUMMARY in solvent B for 9 min, isocratic at 100% B for 2 min, and return to initial CH:CHROMATOGRAPHY_SUMMARY conditions, 5% B for 3 min. A flow rate of 0.5 mL/min was used. Eluent A was CH:CHROMATOGRAPHY_SUMMARY 0.1% formic acid in water and eluent B was 0.1% formic acid in methanol. CH:CHROMATOGRAPHY_SUMMARY Injection volume was 5 µl. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) CH:SOLVENT_A 0.1 % formic acid in water CH:SOLVENT_B 0.1 % formic acid in methanol CH:FLOW_GRADIENT 5 % - 100 % CH:FLOW_RATE 0.5 mL/min CH:COLUMN_TEMPERATURE 40ºC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE Other MS:ION_MODE POSITIVE MS:MS_COMMENTS POS Instrument control and data acquisition was carried out using Xcalibur v.4.3 MS:MS_COMMENTS software. Data treatment, alignment (with a maximum shift of 0.1 min and a mass MS:MS_COMMENTS tolerance of 5 ppm), peak selection, deconvolution, normalization and annotation MS:MS_COMMENTS were performed using Compound Discoverer v3.1 (Thermo Fisher Scientific, Bremen, MS:MS_COMMENTS Germany). MS:MS_RESULTS_FILE ST003016_AN004949_Results.txt UNITS:Peak Area Has m/z:Neutral masses Has RT:Yes RT units:Minutes #END