#METABOLOMICS WORKBENCH gdpu_kjh_20231204_091004 DATATRACK_ID:4487 STUDY_ID:ST002997 ANALYSIS_ID:AN004923 PROJECT_ID:PR001867 VERSION 1 CREATED_ON December 6, 2023, 12:05 am #PROJECT PR:PROJECT_TITLE SZDP Metabolomic studies PR:PROJECT_TYPE LC_MS PR:PROJECT_SUMMARY A metabolomics study was conducted to elucidate the metabolic mechanism of PR:PROJECT_SUMMARY Shenzao dripping pill against myocardial ischemia. PR:INSTITUTE Guangdong Pharmaceutical University PR:DEPARTMENT Engineering & Technology Research Center for Chinese Materia Medica Quality of PR:DEPARTMENT the Universities of Guangdong Province PR:LABORATORY Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State PR:LABORATORY Administration of Traditional Chinese Medicine PR:LAST_NAME Kuang PR:FIRST_NAME Jiehui PR:ADDRESS Waihuan East Road, 280, Guangzhou, Guangdong, 510006, China PR:EMAIL kuang_jieh@163.com PR:PHONE (020)-222-3333 PR:FUNDING_SOURCE This research was funded by the Project of the Science and Technology Plan PR:FUNDING_SOURCE Project of Guangzhou (No. 201803010115), the National Natural Science Foundation PR:FUNDING_SOURCE of China (No. 82173972), and the National Major #STUDY ST:STUDY_TITLE Metabolomic analysis reveal mechanisms of Shenzao dripping pill against ST:STUDY_TITLE myocardial ischemia ST:STUDY_SUMMARY This study employed the advanced Ultra-high performance liquid ST:STUDY_SUMMARY chromatography-quadruple-Exactive Orbitrap mass spectrometry technology to ST:STUDY_SUMMARY detect biomarkers related to myocardial ischemia in rats. Additionally, it aimed ST:STUDY_SUMMARY to investigate the effect of the Shenzao dripping pill on rectifying metabolic ST:STUDY_SUMMARY disorders and preserving the metabolites in dynamic balance during myocardial ST:STUDY_SUMMARY ischemia, analyze potential metabolic pathways, and elucidate the mechanism of ST:STUDY_SUMMARY action of SZDP through the lens of metabolomics. ST:INSTITUTE Guangdong Pharmaceutical University ST:DEPARTMENT Engineering & Technology Research Center for Chinese Materia Medica Quality of ST:DEPARTMENT the Universities of Guangdong Province ST:LABORATORY Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State ST:LABORATORY Administration of Traditional Chinese Medicine ST:LAST_NAME Kuang ST:FIRST_NAME Jiehui ST:ADDRESS Waihuan East Road, 280, Guangzhou, Guangdong, 510006, China ST:EMAIL kuang_jieh@163.com ST:STUDY_TYPE Metabolomic analysis on serum samples of SZDP against MI ST:PHONE (020)-222-3333 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 #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 - M1 Treatment:Coronary ligation surgery RAW_FILE_NAME=M1.raw SUBJECT_SAMPLE_FACTORS - M2 Treatment:Coronary ligation surgery RAW_FILE_NAME=M2.raw SUBJECT_SAMPLE_FACTORS - M3 Treatment:Coronary ligation surgery RAW_FILE_NAME=M3.raw SUBJECT_SAMPLE_FACTORS - M4 Treatment:Coronary ligation surgery RAW_FILE_NAME=M4.raw SUBJECT_SAMPLE_FACTORS - M5 Treatment:Coronary ligation surgery RAW_FILE_NAME=M5.raw SUBJECT_SAMPLE_FACTORS - M6 Treatment:Coronary ligation surgery RAW_FILE_NAME=M6.raw SUBJECT_SAMPLE_FACTORS - M7 Treatment:Coronary ligation surgery RAW_FILE_NAME=M7.raw SUBJECT_SAMPLE_FACTORS - M8 Treatment:Coronary ligation surgery RAW_FILE_NAME=M8.raw SUBJECT_SAMPLE_FACTORS - M9 Treatment:Coronary ligation surgery RAW_FILE_NAME=M9.raw SUBJECT_SAMPLE_FACTORS - S1 Treatment:Sham RAW_FILE_NAME=S1.raw SUBJECT_SAMPLE_FACTORS - S2 Treatment:Sham RAW_FILE_NAME=S2.raw SUBJECT_SAMPLE_FACTORS - S3 Treatment:Sham RAW_FILE_NAME=S3.raw SUBJECT_SAMPLE_FACTORS - S4 Treatment:Sham RAW_FILE_NAME=S4.raw SUBJECT_SAMPLE_FACTORS - S5 Treatment:Sham RAW_FILE_NAME=S5.raw SUBJECT_SAMPLE_FACTORS - S6 Treatment:Sham RAW_FILE_NAME=S6.raw SUBJECT_SAMPLE_FACTORS - S7 Treatment:Sham RAW_FILE_NAME=S7.raw SUBJECT_SAMPLE_FACTORS - S8 Treatment:Sham RAW_FILE_NAME=S8.raw SUBJECT_SAMPLE_FACTORS - SZ1 Treatment:SZDP-H RAW_FILE_NAME=SZ1.raw SUBJECT_SAMPLE_FACTORS - SZ2 Treatment:SZDP-H RAW_FILE_NAME=SZ2.raw SUBJECT_SAMPLE_FACTORS - SZ3 Treatment:SZDP-H RAW_FILE_NAME=SZ3.raw SUBJECT_SAMPLE_FACTORS - SZ4 Treatment:SZDP-H RAW_FILE_NAME=SZ4.raw SUBJECT_SAMPLE_FACTORS - SZ5 Treatment:SZDP-H RAW_FILE_NAME=SZ5.raw SUBJECT_SAMPLE_FACTORS - SZ6 Treatment:SZDP-H RAW_FILE_NAME=SZ6.raw SUBJECT_SAMPLE_FACTORS - SZ7 Treatment:SZDP-H RAW_FILE_NAME=SZ7.raw SUBJECT_SAMPLE_FACTORS - SZ8 Treatment:SZDP-H RAW_FILE_NAME=SZ8.raw #COLLECTION CO:COLLECTION_SUMMARY Blood was collected from abdominal aorta, and the serum samples were obtained by CO:COLLECTION_SUMMARY separating the supernatant from the blood centrifugation with 3,500 rpm for 10 CO:COLLECTION_SUMMARY min after 30 min of coagulation and stored in the refrigerator at −80°C CO:COLLECTION_SUMMARY immediately. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Rats in the model and sham groups were given a solution of 0.5% CMC-Na; in the TR:TREATMENT_SUMMARY high dose of the SZDP group was administered 500 mg/kg SZDP. All rats were TR:TREATMENT_SUMMARY subjected to intragastric administration once daily. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY In each 200 μL aliquot of serum sample, 600 μL of acetonitrile was added and SP:SAMPLEPREP_SUMMARY the mixture was thoroughly mixed for a duration of 1 minute. Then, the mixture SP:SAMPLEPREP_SUMMARY was subjected to centrifugation at 13000 rpm and 4 °C for a period of 15 min. SP:SAMPLEPREP_SUMMARY The supernatant was collected and evaporated to dryness with nitrogen. Methanol SP:SAMPLEPREP_SUMMARY (100 μL) was added, and the reconstituted mixture was vortexed and centrifuged SP:SAMPLEPREP_SUMMARY at 13000 rpm and a temperature of 4 °C for 15 min. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT 0–2 minutes, 5% B; 2–5 minutes, 5%–50% B; 5–6 minutes, 50% B; 6–18 CH:FLOW_GRADIENT minutes, 50%–100% B. CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 40 °C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Mass spectrometry was set to positive and negative ion switching mode using an MS:MS_COMMENTS electrospray ionization source operated at 350 °C. The specific MS conditions MS:MS_COMMENTS were established: the sheath gas flow rate was 55 arb. Units; the auxiliary gas MS:MS_COMMENTS flow rate was 15 arb. Units with a temperature of 200 °C; the electrospray MS:MS_COMMENTS voltage was 3.5 kV in positive ion mode and -3.8 kV in negative ion mode. MS:MS_RESULTS_FILE ST002997_AN004923_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END