#METABOLOMICS WORKBENCH houyuanlong2020_20201209_194452 DATATRACK_ID:2347 STUDY_ID:ST001623 ANALYSIS_ID:AN002658 PROJECT_ID:000000 VERSION 1 CREATED_ON December 11, 2020, 10:21 am #PROJECT PR:PROJECT_TITLE Raffinose underlies diet-dependent epithelial response to stress PR:PROJECT_TYPE Targeted metabolomics analysis with raffinose-rich diet in mouse ileum PR:PROJECT_SUMMARY The raffinose-rich diet was supplemented with raffinose, a significant increase PR:PROJECT_SUMMARY in ileum fructose in stressed mice could be observed. PR:INSTITUTE China Pharmaceutical University PR:DEPARTMENT School of Medicine PR:LABORATORY Metabonomics PR:LAST_NAME Hou PR:FIRST_NAME Yuanlong PR:ADDRESS tongjiaxiang, nanjing, jiangsu, 210000, China PR:EMAIL jian2103@163.com PR:PHONE 18851105337 PR:CONTRIBUTORS Xiao Zheng Haiping Hao #STUDY ST:STUDY_TITLE Targeted metabolomics analysis with raffinose-rich diet in mouse ileum ST:STUDY_TYPE Analysis of fructose level in ileum of stressed mouse fed with raffinose-rich ST:STUDY_TYPE diet ST:STUDY_SUMMARY Purified diet (AIN-93G) supplemented with raffinose was prepared. The mice were ST:STUDY_SUMMARY maintained on the separate diet for at least 1 week before the initiation of ST:STUDY_SUMMARY experiment.Chronic restraint stress (RS) in mice was performed 14 days.After ST:STUDY_SUMMARY sacrifice, the ileum of mice were used to analysis. ST:INSTITUTE China Pharmaceutical University ST:DEPARTMENT School of Medicine ST:LABORATORY Metabonomics ST:LAST_NAME Hou ST:FIRST_NAME Yuanlong ST:ADDRESS tongjiaxiang, nanjing, jiangsu, 210000, China ST:EMAIL jian2103@163.com ST:PHONE 18851105337 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Female #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 - S1 Treatment:Stress RAW_FILE_NAME=20191205HYL-INESTINAL-S1 SUBJECT_SAMPLE_FACTORS - S2 Treatment:Stress RAW_FILE_NAME=20191205HYL-INESTINAL-S2 SUBJECT_SAMPLE_FACTORS - S3 Treatment:Stress RAW_FILE_NAME=20191205HYL-INESTINAL-S3 SUBJECT_SAMPLE_FACTORS - S4 Treatment:Stress RAW_FILE_NAME=20191205HYL-INESTINAL-S4 SUBJECT_SAMPLE_FACTORS - C1 Treatment:Con RAW_FILE_NAME=20191205HYL-INESTINAL-C1 SUBJECT_SAMPLE_FACTORS - C2 Treatment:Con RAW_FILE_NAME=20191205HYL-INESTINAL-C2 SUBJECT_SAMPLE_FACTORS - C3 Treatment:Con RAW_FILE_NAME=20191205HYL-INESTINAL-C3 SUBJECT_SAMPLE_FACTORS - C4 Treatment:Con RAW_FILE_NAME=20191205HYL-INESTINAL-C4 SUBJECT_SAMPLE_FACTORS - R1 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R1 SUBJECT_SAMPLE_FACTORS - R3 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R3 SUBJECT_SAMPLE_FACTORS - R4 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R4 SUBJECT_SAMPLE_FACTORS - R5 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R5 SUBJECT_SAMPLE_FACTORS - R6 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R6 SUBJECT_SAMPLE_FACTORS - R7 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R7 SUBJECT_SAMPLE_FACTORS - R8 Treatment:Raffinose RAW_FILE_NAME=20191205HYL-INTESTINAL-R8 #COLLECTION CO:COLLECTION_SUMMARY jejunum tissue (30 mg) was transferred to homogenizer (pre-cooled at -20 ℃) CO:COLLECTION_SUMMARY and 100 µL of H2O (pre-cooled at 4℃) was added for homogenate for 1 min. 1 CO:COLLECTION_SUMMARY μg/mL 4-chloro-phenylalanine and 800 µL methanol solution were added for CO:COLLECTION_SUMMARY protein precipitation and vortexed for 10 min. The sample lysates were CO:COLLECTION_SUMMARY centrifuged at 30000g for 10 min at 4 °C, and dried under vacuum. It was CO:COLLECTION_SUMMARY reconstituted with 100 μL of methanol: water solution (50:50, v/v) before CO:COLLECTION_SUMMARY analysis. CO:SAMPLE_TYPE Intestine #TREATMENT TR:TREATMENT_SUMMARY PD was supplemented with raffinose, a significant increase in intestinal TR:TREATMENT_SUMMARY fructose in stressed mice could be observed. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Briefly, tissue (30 mg) was transferred to homogenizer (pre-cooled at -20 ℃) SP:SAMPLEPREP_SUMMARY and 100 µL of H2O (pre-cooled at 4℃) was added for homogenate for 1 min. 1 SP:SAMPLEPREP_SUMMARY μg/mL 4-chloro-phenylalanine and 800 µL methanol solution were added for SP:SAMPLEPREP_SUMMARY protein precipitation and vortexed for 10 min. The sample lysates were SP:SAMPLEPREP_SUMMARY centrifuged at 30000g for 10 min at 4 °C, and dried under vacuum. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity H-Class CH:COLUMN_NAME Waters Acquity BEH Amide (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Xevo G2-XS, Waters MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS MS MassLynx v4.1 Quanlynx v4.1 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS relative peak area MS_METABOLITE_DATA_START Samples C1 C2 C3 C4 S1 S2 S3 S4 R1 R3 R4 R5 R6 R7 R8 Factors Treatment:Con Treatment:Con Treatment:Con Treatment:Con Treatment:Stress Treatment:Stress Treatment:Stress Treatment:Stress Treatment:Raffinose Treatment:Raffinose Treatment:Raffinose Treatment:Raffinose Treatment:Raffinose Treatment:Raffinose Treatment:Raffinose Fructose 0.0347609 0.040259 0.04000952 0.0364934 0.025058 0.02226 0.013987 0.01046851 0.07282454 0.04740064 0.05582034 0.0831141 0.07902627 0.07224646 0.05783501 Glucose 0.0119652 0.0275589 0.01848641 0.0119649 0.04606187 0.03815228 0.0118872 0.0306539 0.02514915 0.01567248 0.04050696 0.01958676 0.03802045 0.0201069 0.02894439 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Fructose Glucose METABOLITES_END #END