#METABOLOMICS WORKBENCH FernandezGarcia_M_20221004_151220 DATATRACK_ID:3490 STUDY_ID:ST003970 ANALYSIS_ID:AN006536 PROJECT_ID:PR002486 VERSION 1 CREATED_ON June 13, 2025, 5:09 am #PROJECT PR:PROJECT_TITLE Evaluation of Babesia divergens metabolism PR:PROJECT_SUMMARY The project aims to evaluate the metabolism of the piroplasmid apicomplexa PR:PROJECT_SUMMARY Babesia divergens through metabolomics and complementary functional assays PR:PROJECT_SUMMARY involving metabolites. PR:INSTITUTE Centro Nacional de Microbiología CNM-ISCIII PR:LAST_NAME Estrella PR:FIRST_NAME Montero PR:ADDRESS Ctra. de Pozuelo, 28, 28222 Majadahonda, Madrid, España PR:EMAIL estrella.montero@isciii.es PR:PHONE +34 918 22 36 57 #STUDY ST:STUDY_TITLE Comprehensive multiplatform mass spectrometry-based metabolomics unveils ST:STUDY_TITLE insights into Babesia divergens metabolism. ST:STUDY_SUMMARY Babesia divergens is a zoonotic parasite belonging to the apicomplexa phyla ST:STUDY_SUMMARY causing human babesiosis. However, its metabolism remains poorly understood as ST:STUDY_SUMMARY scarce biochemical studies have been reported, and no global metabolic network ST:STUDY_SUMMARY reconstructions have been reported up to date. Comprehensive understanding of ST:STUDY_SUMMARY the B. divergens metabolism could reveal novel therapeutic targets. We performed ST:STUDY_SUMMARY a multiplatform analysis combining targeted and non-targeted mass spectrometry ST:STUDY_SUMMARY based metabolomics of samples (pellets, supernatants) obtained from Babesia ST:STUDY_SUMMARY divergens-infected erythrocyte cultures and non-infected culture controls, ST:STUDY_SUMMARY acquired with CE-ESI(+)/MS, GC-CI(+)/MS, LC-ESI(+)/MS, LC-ESI(-)/MS (all ST:STUDY_SUMMARY non-targeted analyses) and LC-ESI(-)/MS (targeted analysis). Profound metabolite ST:STUDY_SUMMARY level changes occur in both the B. divergens-RBC system and its supernatant ST:STUDY_SUMMARY compared to controls, including those related with central carbon metabolism, ST:STUDY_SUMMARY pyrimidine nucleotide biosynthesis, redox metabolism, glycerophospholipid and ST:STUDY_SUMMARY sphingolipid metabolism, amino acid metabolism and nucleotide metabolism. This ST:STUDY_SUMMARY resource serves as a comprehensive dataset describing how the metabolic ST:STUDY_SUMMARY alterations caused by the intraerythrocytic stage of Babesia divergens are ST:STUDY_SUMMARY reflected in different metabolite profiles across multiple ST:STUDY_SUMMARY mass-spectrometry-based platforms. ST:INSTITUTE CEMBIO ST:LAST_NAME Fernández-García ST:FIRST_NAME Miguel ST:ADDRESS Universidad San Pablo-CEU, Urbanización Montepríncipe, 28925, Alcorcón, ST:ADDRESS Madrid, Spain ST:EMAIL mig.fernandez.ce@ceindo.ceu.es ST:PHONE 690090778 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #FACTORS #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 iRBC1 iRBC1_CE-TOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iRBC1_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC2 iRBC2_CE-TOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iRBC2_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_CE-TOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iRBC3_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_CE-TOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iRBC4_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_CE-TOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iRBC5_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC1 uRBC1_CE-TOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uRBC1_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC2 uRBC2_CE-TOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uRBC2_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_CE-TOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uRBC3_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_CE-TOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uRBC4_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_CE-TOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uRBC5_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS1 iS1_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iS1_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS2 iS2_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iS2_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS3 iS3_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iS3_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS4 iS4_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iS4_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS5 iS5_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=iS5_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS1 uS1_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uS1_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS2 uS2_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uS2_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS3 uS3_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uS3_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS4 uS4_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uS4_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS5 uS5_CE-TOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=CE-TOF-MS; RAW_FILE_NAME=uS5_CE-TOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC1 iRBC1_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iRBC1_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC2 iRBC2_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iRBC2_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iRBC3_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iRBC4_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iRBC5_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC1 uRBC1_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uRBC1_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC2 uRBC2_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uRBC2_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uRBC3_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uRBC4_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uRBC5_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS1 iS1_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iS1_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS2 iS2_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iS2_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS3 iS3_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iS3_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS4 iS4_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iS4_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iS5 iS5_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=iS5_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS2 uS2_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uS2_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS3 uS3_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uS3_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS4 uS4_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uS4_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS uS5 uS5_GC-QTOF-MS Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=GC-QTOF-MS; RAW_FILE_NAME=uS5_GC-QTOF-MS.mzML SUBJECT_SAMPLE_FACTORS iRBC1 iRBC1_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iRBC1_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iRBC2 iRBC2_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iRBC2_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iRBC3_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iRBC4_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iRBC5_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uRBC1 uRBC1_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uRBC1_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uRBC2 uRBC2_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uRBC2_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uRBC3_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uRBC4_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uRBC5_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iS1 iS1_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iS1_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iS2 iS2_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iS2_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iS3 iS3_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iS3_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iS4 iS4_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iS4_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iS5 iS5_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=iS5_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uS1 uS1_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uS1_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uS2 uS2_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uS2_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uS3 uS3_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uS3_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uS4 uS4_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uS4_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS uS5 uS5_LC-QTOF-MSpos Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QTOF-MSpos; RAW_FILE_NAME=uS5_LC-QTOF-MSpos.mzML SUBJECT_SAMPLE_FACTORS iRBC1 iRBC1_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=iRBC1_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS iRBC2 iRBC2_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=iRBC2_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=iRBC3_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=iRBC4_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=iRBC5_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS uRBC1 uRBC1_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=uRBC1_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=uRBC3_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=uRBC4_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_LC-QTOF-MSneg Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QTOF-MSneg; RAW_FILE_NAME=uRBC5_LC-QTOF-MSneg.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iRBC3_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iRBC4_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iRBC5_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uRBC2 uRBC2_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uRBC2_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uRBC3_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uRBC4_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uRBC5_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iS1 iS1_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iS1_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iS2 iS2_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iS2_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iS3 iS3_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iS3_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iS4 iS4_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iS4_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iS5 iS5_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=iS5_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uS1 uS1_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uS1_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uS2 uS2_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uS2_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uS3 uS3_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uS3_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uS4 uS4_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uS4_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS uS5 uS5_LC-QqQ-MS A Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS A; RAW_FILE_NAME=uS5_LC-QqQ-MS A.mzML SUBJECT_SAMPLE_FACTORS iRBC1 iRBC1_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iRBC1_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iRBC3 iRBC3_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iRBC3_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iRBC4 iRBC4_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iRBC4_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iRBC5 iRBC5_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iRBC5_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uRBC2 uRBC2_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uRBC2_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uRBC3 uRBC3_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uRBC3_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uRBC4 uRBC4_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uRBC4_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uRBC5 uRBC5_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uRBC5_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iS1 iS1_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iS1_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iS2 iS2_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iS2_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iS3 iS3_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iS3_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iS4 iS4_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iS4_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS iS5 iS5_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=iS5_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uS1 uS1_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uS1_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uS2 uS2_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uS2_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uS3 uS3_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uS3_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uS4 uS4_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uS4_LC-QqQ-MS B.mzML SUBJECT_SAMPLE_FACTORS uS5 uS5_LC-QqQ-MS B Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Analytical platform=LC-QqQ-MS B; RAW_FILE_NAME=uS5_LC-QqQ-MS B.mzML #COLLECTION CO:COLLECTION_SUMMARY Human A+ erythrocytes were obtained from healthy volunteer donors at the Blood CO:COLLECTION_SUMMARY Transfusion Center in Madrid, Spain. Donors provided written informed consent CO:COLLECTION_SUMMARY for the use of their blood in research. All procedures were approved by the CO:COLLECTION_SUMMARY center and carried out in accordance with institutional and regulatory CO:COLLECTION_SUMMARY guidelines. Asynchronous cultures of Babesia divergens (Bd Rouen 1986 strain) CO:COLLECTION_SUMMARY were maintained using human A+ erythrocytes at 5% hematocrit in RPMI 1640 medium CO:COLLECTION_SUMMARY (Gibco, Grand Island, NY, USA), supplemented with 10% human serum (The CO:COLLECTION_SUMMARY Interstate Companies, Memphis, TN, USA), 7.5% (w/v) sodium bicarbonate (Lonza CO:COLLECTION_SUMMARY Group Ltd., Basel, Switzerland), and 50 μg/mL hypoxanthine (Sigma-Aldrich, CO:COLLECTION_SUMMARY St. Louis, MO, USA). Control flasks containing uninfected erythrocytes were CO:COLLECTION_SUMMARY incubated under the same conditions using identical medium. All cultures were CO:COLLECTION_SUMMARY maintained at 37 °C in a humidified atmosphere with 5% CO₂. Samples were CO:COLLECTION_SUMMARY collected from independent asynchronous cultures of B. divergens at CO:COLLECTION_SUMMARY approximately 40% parasitemia. Infected cultures were centrifuged at CO:COLLECTION_SUMMARY 600 × g for 5 minutes at 4 °C using fixed-angle rotors to pellet the CO:COLLECTION_SUMMARY cells (iRBC). The pellets, containing a mixture of infected and uninfected CO:COLLECTION_SUMMARY erythrocytes, were washed and centrifuged three times (2,000 × g, 5 CO:COLLECTION_SUMMARY minutes, 4 °C), resuspended in RPMI, and kept on ice. Supernatants (iS) were CO:COLLECTION_SUMMARY obtained from infected cultures by sequential centrifugation at 2,000 × g CO:COLLECTION_SUMMARY and 8,000 × g (5 minutes each, 4 °C), and stored at –80 °C. Control CO:COLLECTION_SUMMARY samples (uRBC and uS) from uninfected cultures were processed in parallel using CO:COLLECTION_SUMMARY the same protocol. Detailed description of sample collection can be found at CO:COLLECTION_SUMMARY DOI: 10.1007/978-1-0716-1681-9_13 CO:SAMPLE_TYPE Red blood cells and culture media #TREATMENT TR:TREATMENT_SUMMARY Since the study was aimed to describe changes ocurring upon an in vitro model of TR:TREATMENT_SUMMARY a natural disease, no treatment was performed. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolite extraction was carried out using a multi-step procedure designed to SP:SAMPLEPREP_SUMMARY rapidly quench metabolism, promote cell lysis, and enhance metabolite recovery. SP:SAMPLEPREP_SUMMARY For intracellular extraction, 800 μL of cold (–20 °C) methanol (Thermo SP:SAMPLEPREP_SUMMARY Fisher Scientific, Loughborough, UK) was added to 200 μL of either infected SP:SAMPLEPREP_SUMMARY (iRBC) or uninfected (uRBC) red blood cells. Samples underwent two freeze–thaw SP:SAMPLEPREP_SUMMARY cycles by immersion in liquid nitrogen for 10 minutes, followed by thawing on SP:SAMPLEPREP_SUMMARY ice and light vortexing. After centrifugation at 5,725 × g for 5 minutes at SP:SAMPLEPREP_SUMMARY 4 °C, the supernatants were collected. The residual pellets were re-extracted SP:SAMPLEPREP_SUMMARY twice with 400 μL of cold methanol per step, using the same freeze–thaw and SP:SAMPLEPREP_SUMMARY centrifugation conditions. All supernatants from the same sample were pooled and SP:SAMPLEPREP_SUMMARY stored at –80 °C until analysis. Extracellular samples (iS and uS) were SP:SAMPLEPREP_SUMMARY processed based on the analytical platform. For untargeted CE-TOF/MS, SP:SAMPLEPREP_SUMMARY GC-QTOF/MS, and LC-QTOF/MS analyses, cold acetonitrile (Thermo Fisher SP:SAMPLEPREP_SUMMARY Scientific, Loughborough, UK) was used in a 1:4 (v/v) sample-to-solvent ratio. SP:SAMPLEPREP_SUMMARY For targeted LC-QqQ/MS profiling, cold methanol was used in a 1:3 (v/v) ratio. SP:SAMPLEPREP_SUMMARY Following quenching and extraction, supernatants were filtered through SP:SAMPLEPREP_SUMMARY 0.22 μm MS® Nylon syringe filters (Membrane Solutions, Plano, USA) and SP:SAMPLEPREP_SUMMARY stored at –80 °C. For CE-TOF/MS analysis, 250 μL of each extract (iRBC, SP:SAMPLEPREP_SUMMARY uRBC, iS, and uS) was dried under vacuum and reconstituted in 50 μL of CE/MS SP:SAMPLEPREP_SUMMARY sample solution prepared by dissolving methionine sulfone (internal standard; SP:SAMPLEPREP_SUMMARY Sigma-Aldrich, Steinheim, Germany) at 0.2 mM in Milli-Q water with 0.1 M SP:SAMPLEPREP_SUMMARY formic acid (Sigma-Aldrich). After vortexing for 1 minute and centrifugation at SP:SAMPLEPREP_SUMMARY 12,600 × g for 15 minutes at 4 °C, the supernatants were used for analysis. SP:SAMPLEPREP_SUMMARY GC-QTOF/MS samples were prepared by adding 10 μL of ethoxymation solution and SP:SAMPLEPREP_SUMMARY 12 μL of internal standard to 240 μL of each sample. The ethoxymation SP:SAMPLEPREP_SUMMARY reagent was anhydrous pyridine containing 19 mg·mL⁻¹ O-ethoxyamine SP:SAMPLEPREP_SUMMARY (Sigma-Aldrich, Steinheim, Germany), and the internal standard solution SP:SAMPLEPREP_SUMMARY contained 10 mg·L⁻¹ meso-erythritol in Milli-Q water. After vacuum drying, SP:SAMPLEPREP_SUMMARY derivatization was performed automatically using an MPS autosampler. SP:SAMPLEPREP_SUMMARY O-ethyloxime derivatives were formed by incubating the extracts for 90 minutes SP:SAMPLEPREP_SUMMARY at 40 °C, followed by silylation of acidic compounds using 42 μL of SP:SAMPLEPREP_SUMMARY N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) with 1% SP:SAMPLEPREP_SUMMARY trimethylchlorosilane (TCMS; Sigma-Aldrich, Germany) for 50 minutes at 40 °C. SP:SAMPLEPREP_SUMMARY For LC-QTOF/MS analysis, 100 μL of each processed extract (iRBC, uRBC, iS, SP:SAMPLEPREP_SUMMARY and uS) was transferred to LC vials after brief vortexing. For LC-QqQ/MS SP:SAMPLEPREP_SUMMARY analysis of polar metabolites, 12 μL of internal standard solution SP:SAMPLEPREP_SUMMARY (50.09 µM p-chlorophenylalanine in Milli-Q water; Sigma-Aldrich, Steinheim, SP:SAMPLEPREP_SUMMARY Germany) were added to 300 μL of each extract — methanol for iRBC/uRBC and SP:SAMPLEPREP_SUMMARY methanol:water (3:1, v/v) for iS/uS. Samples were then evaporated under vacuum, SP:SAMPLEPREP_SUMMARY reconstituted in 60 μL of Milli-Q water with the aid of a 5-minute sonication SP:SAMPLEPREP_SUMMARY bath, and centrifuged at 3,000 × g for 10 minutes at 4 °C before injection. SP:SAMPLEPREP_SUMMARY Detailed protocols of the conditions followed sample preparation can be found at SP:SAMPLEPREP_SUMMARY DOI: 10.1007/978-1-0716-1681-9_13 #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY targeted, LC-MS method for the determination of polar compounds. Ion pair with CH:CHROMATOGRAPHY_SUMMARY tributylamine CH:CHROMATOGRAPHY_TYPE Ion pair CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Agilent RRHD Zorbax Extend C18 column (150 x 2.1mm, 1.8 um) CH:SOLVENT_A 97% water/3% methanol; 15 mM acetic acid; 10 mM tributylamine CH:SOLVENT_B 100% methanol; 15 mM acetic acid; 10 mM tributylamine CH:FLOW_GRADIENT 0%B at 0-2.5 min; increased up to 20%B at 7.5 min; increased up to 45%B at 13 CH:FLOW_GRADIENT min; increased up to 99%B at 20 min; held at 24 min; washing step with CH:FLOW_GRADIENT quaternary pump until 59 min; then 0%B at 59 min CH:FLOW_RATE 0.25 mL/min (until 24 min); 0.2 mL/min at 59 min, gradually increased up to 0.25 CH:FLOW_RATE mL/min at 60 min CH:COLUMN_TEMPERATURE 35 ºC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6460 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS MS method associated with LC-ESI(-)-QqQ-MS Drying gas flow rate = 13 L/min MS:MS_COMMENTS Drying gas temperature = 150 ºC Sheath gas flow rate = 12 L/min Sheath gas MS:MS_COMMENTS temperature = 350 ºC Capillary voltage = 2000 V Nozzle voltage = 500 V MS:MS_COMMENTS Acquisition mode = dynamic MRM cycle time = 650 ms #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Internal standard normalized abundances MS_METABOLITE_DATA_START Samples iRBC1_LC-QqQ-MS B iRBC3_LC-QqQ-MS B iRBC4_LC-QqQ-MS B iRBC5_LC-QqQ-MS B uRBC2_LC-QqQ-MS B uRBC3_LC-QqQ-MS B uRBC4_LC-QqQ-MS B uRBC5_LC-QqQ-MS B iS1_LC-QqQ-MS B iS2_LC-QqQ-MS B iS3_LC-QqQ-MS B iS4_LC-QqQ-MS B iS5_LC-QqQ-MS B uS1_LC-QqQ-MS B uS2_LC-QqQ-MS B uS3_LC-QqQ-MS B uS4_LC-QqQ-MS B uS5_LC-QqQ-MS B Factors Sample source:erythrocyte culture | Sample type:infected RBC Sample source:erythrocyte culture | Sample type:infected RBC Sample source:erythrocyte culture | Sample type:infected RBC Sample source:erythrocyte culture | Sample type:infected RBC Sample source:erythrocyte culture | Sample type:uninfected RBC Sample source:erythrocyte culture | Sample type:uninfected RBC Sample source:erythrocyte culture | Sample type:uninfected RBC Sample source:erythrocyte culture | Sample type:uninfected RBC Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Sample source:erythrocyte culture | Sample type:supernatant from infected RBC Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Sample source:erythrocyte culture | Sample type:supernatant from uninfected RBC Adenosine-5'-diphosphate 1.295639495 1.475642094 0.810869167 8.123518972 6.60222828 0.80565945 0.270266786 0.841886459 1.554189018 4.786601283 1.390273378 1.836552254 2.379621195 NaN NaN NaN NaN NaN Adenosine-5'-monophosphate 916.0601623 1836.929908 1770.760266 4753.675181 70.08916002 35.45754974 31.74724456 35.2664753 707.9736933 896.3743136 678.64687 587.9329609 639.2190349 3.779238483 5.128088422 3.011835874 3.891661603 3.00488041 alpha-Ketoglutaric acid 1893.910954 2893.385887 3686.093286 5559.21689 3703.115592 1902.308874 1072.708661 1939.93056 15556.80299 13442.74486 12802.08146 12561.22627 11610.92035 2112.852183 2148.076687 2134.548538 2201.739964 2056.99972 Asparagine 220.5313815 148.5014221 149.2104342 151.0629274 145.1555111 131.4859204 464.315866 113.1730266 106.3328029 118.1516487 130.0724835 125.0330051 121.5125343 103.5925192 98.49263176 104.0841217 100.2622426 107.5446225 Citric acid // Isocitric acid 63.08737568 55.44391656 68.06711273 67.8612874 341.6961651 93.35240558 84.49357165 80.76702633 38988.90311 57275.4263 49152.15042 45477.56908 49978.82308 57516.08996 79865.61965 76307.58151 77054.16234 41772.18376 Dihydroxyacetone phosphate 70.97325574 48.09123045 42.80281501 139.9331626 1554.226185 296.6109635 362.6291761 225.4865957 449.7149546 943.4060318 972.3181808 800.2311693 707.4073219 19.55081051 36.74624307 50.68855695 25.88979087 53.16016618 Fructose-6-phosphate 22.32361847 16.14441095 9.811866421 7.638338156 236.6563151 385.8062322 1589.1418 336.7166011 15.86532427 16.34487824 18.94405925 17.74810649 16.92343704 0.824816657 0.797584349 1.417635984 1.316520108 0.04931453 Fumaric acid 90.69274749 149.772557 156.5872905 211.8740887 345.2047179 121.2902645 81.1609104 117.1274762 318.9284261 246.1684531 275.2404134 261.852612 250.15678 885.5311675 839.8775295 842.8057281 853.8273634 886.4024748 Glucose-6-phosphate 11.50993617 8.881036881 4.361517369 13.29879338 249.4774057 278.6097186 339.2972818 264.3375087 47.04617702 57.26201054 32.55414993 44.73147907 43.13317597 2.582121188 3.063731831 3.582217149 3.787508667 1.240733167 Glutamic acid 4791.874481 5488.033971 5904.35943 9557.15741 19978.53491 10813.46237 15571.1174 10385.49907 22026.02413 17734.8498 23999.59414 27604.61593 26170.74388 5889.220632 6327.690348 5400.914249 4959.30165 9234.48517 Glutamine 195.8341331 130.6491042 133.0082462 144.6022361 371.2565235 326.5214166 1135.206469 285.1825051 111.3293252 122.9881887 121.6796854 118.8905103 109.6641164 318.0037237 333.2787348 318.5430326 351.6196535 305.7648163 Histidine 57.31204082 47.19564232 48.9423888 52.71657115 54.21612156 46.16039951 147.7419445 42.52766305 38.61247485 46.08236623 37.61191252 42.24872692 38.17560296 37.98308057 33.39566408 36.72261332 40.34598569 43.19462168 Isoleucine 11.60578438 12.02920608 17.21948683 14.2766207 17.15504515 13.51776515 30.31747306 11.37201381 48.51484126 34.25893121 52.41322887 53.1544872 53.43457282 41.48967786 44.56804931 36.3326326 38.47885343 48.30563129 Itaconic acid 265.8911733 2234.792896 2270.946543 17842.69187 35.52698069 15.66228208 48.91327999 38.68291718 221.7829014 315.5668784 418.5470879 367.6676577 326.9596878 131.237128 129.7378873 124.9963247 140.3603613 128.3014019 Lactic acid 9012.856186 8523.440886 9178.518861 10584.15973 5237.816535 6064.140766 16793.02204 5821.374929 12147.71603 8291.407329 9314.435454 12412.0539 10317.87163 4720.449239 6252.712414 3356.1923 2969.295917 6931.066423 Leucine 71.38629385 77.20651273 86.08103848 130.9001918 155.6663312 88.01044499 120.3989625 86.98941228 375.9343746 170.5604066 345.180326 444.5245097 432.3853289 357.2496719 460.6495699 293.307005 240.5138626 477.2131247 Malic acid 8173.913113 11375.76716 12261.291 22851.26178 25723.63983 14090.75236 8057.992732 13670.76188 183567.5422 198132.6485 180399.9129 180992.8357 179449.5594 108356.7952 101598.8139 104395.5914 115617.2818 98770.61891 Methionine 97.07001107 132.4033324 169.0576032 257.7765165 248.6144548 149.9247408 244.2282893 147.4238506 644.1951859 651.7804791 546.2396 594.3846266 561.7473895 398.8053741 470.8884027 445.167696 416.0696218 466.4830502 NAD+ 1229.408338 710.2402003 928.1921899 1121.649358 2549.236978 2291.08749 5278.504709 1871.997422 NaN NaN NaN NaN NaN NaN NaN NaN NaN NaN NADH 0.861733492 5.566238813 6.548553137 25.68299989 0.89343733 0.221914623 NaN 0.569553207 NaN NaN NaN NaN NaN NaN NaN NaN NaN NaN Phenylalanine 1353.858107 1712.723931 1988.608097 3156.732815 4944.640079 2088.788789 2177.61999 2175.265314 3590.502477 487.8857334 3974.053867 6181.77489 5153.904274 4513.879309 5430.649222 3385.641081 2709.958002 13176.88229 Phosphoenolpyruvic acid 3.500215021 3.546713484 2.449043718 16.99003579 25.16001877 5.178522758 NaN 3.552504366 13268.37217 18572.58302 14931.84019 15769.68599 15175.89409 117.662541 50.82572844 121.3000878 130.0088074 83.18880329 Proline 6.572857328 7.090382991 7.293079764 7.806997311 8.311240437 7.635090966 14.84508601 5.747774412 8.18199513 8.289563566 5.750153046 7.306572985 7.48642085 6.268337015 8.078602 6.003236647 7.647121883 5.28882215 Pyruvic acid 9029.763321 8104.441595 9264.208986 9978.048558 767.5736348 526.7027256 944.1003994 538.0447396 23051.54382 19225.92588 18472.57413 17617.42943 16505.91397 1493.872709 1524.001195 1679.347569 1461.622124 1803.679483 Ribose-5-phosphate 6.737838808 4.944616879 0.720632737 3.484567776 24.44130481 26.88539856 80.8600189 20.5485129 16.689974 14.71737202 15.9037017 20.1039647 9.253675611 3.612722264 3.521769181 4.290098418 1.63822383 4.269456891 Sedoheptulose-7-phosphate 15.37496097 8.403065726 2.921743883 6.044257195 20.22155915 36.38822244 132.2609302 29.29119292 5.129538971 5.182965069 4.821252737 7.376818036 4.541093065 0.465351128 0.911630766 0.617541351 0.81303186 NaN Serine 136.9981058 103.0250121 112.2310093 118.3272265 81.37450579 90.86921225 311.3189632 84.18491184 71.82230455 65.05815889 48.44413712 46.61295669 34.13521414 60.07224959 53.83541258 44.31436214 56.21720326 34.95236331 Succinic acid 4865.547221 6871.811624 8266.951382 12726.30499 3860.295451 2301.094712 6217.726086 1998.38561 41235.21003 47309.87183 39497.63169 41326.6943 40257.24911 9856.907943 11268.5153 9034.850182 10474.26145 9966.541775 Threonine 118.7647639 102.8828043 95.78106528 117.8527016 101.4497896 102.8634686 257.5766541 88.34509879 128.2627025 138.1481775 127.6310349 117.3769652 120.8612869 108.1911535 128.628077 110.0115748 127.7166329 115.3503449 Tyrosine 293.6234538 336.583072 375.4324629 613.3350635 816.1268284 398.161272 520.9664329 392.3920627 2066.604675 1076.585983 2130.63295 2359.519304 2211.559423 2296.758509 2614.225956 2082.226907 1671.666753 2787.454042 Xylulose-5-phosphate // Ribulose-5-phosphate 77.46767993 66.97676822 49.98714448 105.1610934 630.0629317 287.4542596 322.9040423 260.2015845 38.15806407 24.16389625 33.74439444 27.4751988 24.08205958 4.647080867 5.269973908 12.74053474 4.501620342 10.87230976 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG COMPOUND ID Molecular feature (transition(s)@RT) transition(s) (first quantifier, second qualifier) RT ΔRT error (%) Adenosine-5'-diphosphate C00008 (426.0 -> 328.0), (426.0 -> 159.0) @14.571 (426.0 -> 328.0), (426.0 -> 159.0) 14.571 5.9 Adenosine-5'-monophosphate C00020 (346.0 -> 79.0), (346.0 -> 97.0) @11.336 (346.0 -> 79.0), (346.0 -> 97.0) 11.336 5.9 alpha-Ketoglutaric acid C00026 (145.0 -> 57.2), (145.0 -> 101.0) @13.463 (145.0 -> 57.2), (145.0 -> 101.0) 13.463 -0.35 Asparagine C00152 (131.0 -> 113.0), (131.0 -> 70.0) @1.372 (131.0 -> 113.0), (131.0 -> 70.0) 1.372 -0.09 Citric acid // Isocitric acid C00158 // C00311 (191.0 -> 111.0), (191.0 -> 87.0) @14.1 (191.0 -> 111.0), (191.0 -> 87.0) 14.1 -1.15 Dihydroxyacetone phosphate C00111 (169.0 -> 97.0), (169.0 -> 79.0) @9.728 (169.0 -> 97.0), (169.0 -> 79.0) 9.728 -0.43 Fructose-6-phosphate C00085 (259.0 -> 97.0), (259.0 -> 79.0) @7.402 (259.0 -> 97.0), (259.0 -> 79.0) 7.402 -0.44 Fumaric acid C00122 (115.0 -> 71.0) @13.845 (115.0 -> 71.0) 13.845 0.52 Glucose-6-phosphate C00668 (259.0 -> 97.0), (259.0 -> 79.0) @6.947 (259.0 -> 97.0), (259.0 -> 79.0) 6.947 -0.14 Glutamic acid C00025 (146.0 -> 102.0), (146.0 -> 128.0) @3.53 (146.0 -> 102.0), (146.0 -> 128.0) 3.53 2.2 Glutamine C00064 (145.1 -> 127.0), (145.1 -> 109.0) @1.381 (145.1 -> 127.0), (145.1 -> 109.0) 1.381 2.4 Histidine C00135 (154.1 -> 93.1), (154.1 -> 137.0) @1.325 (154.1 -> 93.1), (154.1 -> 137.0) 1.325 -2.1 Isoleucine C00407 (130.0 -> 45.0) @2.228 (130.0 -> 45.0) 2.228 -0.67 Itaconic acid C00490 (129.0 -> 85.1), (129.0 -> 41.3) @12.828 (129.0 -> 85.1), (129.0 -> 41.3) 12.828 -5.4 Lactic acid C00186 (89.0 -> 43.3), (89.0 -> 45.3) @6.986 (89.0 -> 43.3), (89.0 -> 45.3) 6.986 0.25 Leucine C00123 (130.0 -> 45.0) @2.388 (130.0 -> 45.0) 2.388 1.3 Malic acid C00149 (133.0 -> 115.0), (133.0 -> 71.1) @12.885 (133.0 -> 115.0), (133.0 -> 71.1) 12.885 1.4 Methionine C00073 (148.0 -> 47.2), (148.0 -> 100.0) @1.854 (148.0 -> 47.2), (148.0 -> 100.0) 1.854 -0.14 NAD+ C00003 (662.1 -> 540.0), (662.1 -> 327.9) @8.886 (662.1 -> 540.0), (662.1 -> 327.9) 8.886 124.4086128 NADH C00004 (664.4 -> 78.9), (664.4 -> 407.9) @14.509 (664.4 -> 78.9), (664.4 -> 407.9) 14.509 NaN Phenylalanine C00079 (164.1 -> 147.0), (164.1 -> 103.0) @4.788 (164.1 -> 147.0), (164.1 -> 103.0) 4.788 114.4054385 Phosphoenolpyruvic acid C00074 (167.0 -> 79.0) @14.503 (167.0 -> 79.0) 14.503 3 Proline C00148 (114.0 -> 68.1), (114.0 -> 45.2) @1.446 (114.0 -> 68.1), (114.0 -> 45.2) 1.446 0.9181718 Pyruvic acid C00022 (87.0 -> 43.2) @8.988 (87.0 -> 43.2) 8.988 2175.206634 Ribose-5-phosphate C00117 (229.0 -> 97.0), (229.0 -> 79.0) @7.295 (229.0 -> 97.0), (229.0 -> 79.0) 7.295 0.05 Sedoheptulose-7-phosphate C05382 (289.0 -> 96.9), (289.0 -> 79.0) @7.35 (289.0 -> 96.9), (289.0 -> 79.0) 7.35 -2.7 Serine C00065 (104.0 -> 74.1) @1.376 (104.0 -> 74.1) 1.376 -0.17 Succinic acid C00042 (117.0 -> 73.1), (117.0 -> 99.0) @12.148 (117.0 -> 73.1), (117.0 -> 99.0) 12.148 -1.64 Threonine C00188 (118.0 -> 74.1) @1.403 (118.0 -> 74.1) 1.403 0.27 Tyrosine C00082 (180.0 -> 119.0), (180.0 -> 93.0) @2.259 (180.0 -> 119.0), (180.0 -> 93.0) 2.259 -0.78 Xylulose-5-phosphate // Ribulose-5-phosphate C00231 // C01101 (229.0 -> 79.0, 229.0 -> 97.0) @8.22 (229.0 -> 79.0, 229.0 -> 97.0) 8.22 ≈3.80 METABOLITES_END #END