#METABOLOMICS WORKBENCH FernandezGarcia_M_20221004_151220 DATATRACK_ID:3490 STUDY_ID:ST003970 ANALYSIS_ID:AN006531 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 Cromatography summary. Non-targeted GC-MS method adapted for the determination CH:CHROMATOGRAPHY_SUMMARY of metabolites with acidic hydrogens through two-step derivatization CH:CHROMATOGRAPHY_SUMMARY (ethoxymation and trimethylsilylation). CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Macherey Nagel Optima 726 (60 m x 0.25 mm, 0.25 um) CH:SOLVENT_A - CH:SOLVENT_B - CH:FLOW_GRADIENT - CH:FLOW_RATE 1.3 mL/min CH:COLUMN_TEMPERATURE 80º C for 1 min. Then, increase at 20 ºC/min until 200 ºC. Then, increase at CH:COLUMN_TEMPERATURE 5 ºC/min until 235 ºC. Then, increase at 3 ºC/min until 270 ºC. Finally, CH:COLUMN_TEMPERATURE increase at 20 ºC/min until 320 ºC, which was held for 1 min. #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 7200 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS MS method associated with GC-CI-MS Chemical ionization Gas = methane Gas MS:MS_COMMENTS pressure = 20% Source temperature = 150 ºC Source polarity = positive MS:MS_COMMENTS Acquisition method = full-MS at Extended Dynamic Range (2 GH) Mass range = 70-90 MS:MS_COMMENTS m/z Acquisition rate = 3.33 spectra/s #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS normalized (total useful signal) abundances MS_METABOLITE_DATA_START Samples iRBC1_GC-QTOF-MS iRBC2_GC-QTOF-MS iRBC3_GC-QTOF-MS iRBC4_GC-QTOF-MS iRBC5_GC-QTOF-MS uRBC1_GC-QTOF-MS uRBC2_GC-QTOF-MS uRBC3_GC-QTOF-MS uRBC4_GC-QTOF-MS uRBC5_GC-QTOF-MS iS1_GC-QTOF-MS iS2_GC-QTOF-MS iS3_GC-QTOF-MS iS4_GC-QTOF-MS iS5_GC-QTOF-MS uS2_GC-QTOF-MS uS3_GC-QTOF-MS uS4_GC-QTOF-MS uS5_GC-QTOF-MS 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: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: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 1-Methylhydantoin // Dihidrouracil 0.000130 0.0001154 0.0001164 0.0001175 0.0001210 0.0000786 0.0000720 0.0000725 0.0000746 0.0000778 1.32578E-05 4.04164E-05 2.27271E-05 2.32159E-05 2.54989E-05 9.70908E-06 1.07767E-05 3.84858E-05 6.5564E-06 2-Hydroxybutyric acid 0.000046 0.0000515 0.0000517 0.0000547 0.0000562 0.0000580 0.0000571 0.0000644 0.0000577 0.0000511 0.000347993 0.000263442 0.000343527 0.000325018 0.000307555 0.000542098 0.000465112 0.000449692 0.000452303 2-Hydroxyglutaric acid 0.000447 0.0004326 0.0004035 0.0003989 0.0004088 0.0004847 0.0004960 0.0004985 0.0005364 0.0003984 NaN 5.28963E-06 6.93795E-06 NaN 1.01184E-05 NaN NaN NaN NaN Alanine 0.008342 0.0091101 0.0092204 0.0099838 0.0087197 0.0075992 0.0077756 0.0071895 0.0070350 0.0063572 0.015086746 0.012578989 0.013646168 0.007589391 0.013562488 0.009994675 0.010164996 0.010244336 0.009339818 Aminomalonic acid 0.000163 0.0001315 0.0001143 0.0001290 0.0001460 0.0001321 0.0001305 0.0001616 0.0001525 0.0001658 3.25079E-05 3.68743E-05 6.26377E-05 3.1257E-05 5.52165E-05 2.39982E-05 4.40349E-05 3.67306E-05 3.23079E-05 Arabitol // Ribitol 0.000045 0.0000539 0.0000456 0.0000500 0.0000485 0.0000763 0.0000789 0.0000613 0.0000596 0.0000757 3.23806E-05 3.50809E-05 5.13025E-05 4.57175E-05 4.60373E-05 7.49997E-05 7.75209E-05 6.52573E-05 5.85363E-05 beta-Glycerophosphate 0.000191 0.0003069 0.0002934 0.0003030 0.0002378 0.0002241 0.0002490 0.0002102 0.0002109 0.0001432 4.20889E-05 5.24674E-05 4.42215E-05 4.16911E-05 6.07821E-05 NaN 3.3155E-06 1.58678E-06 2.69433E-06 C14:0 Myristic acid 0.000799 0.0008896 0.0008749 0.0007986 0.0008811 0.0009150 0.0007151 0.0007718 0.0008189 0.0009143 0.000251199 0.000365166 0.000271118 0.000250893 0.000364659 0.000146089 0.000266665 0.000385008 0.000251624 C15:0 Pentadecanoic acid 0.000530 0.0005910 0.0005731 0.0005830 0.0006463 0.0007371 0.0005437 0.0005832 0.0006009 0.0006670 0.000161652 0.000222621 0.000156801 0.000126836 0.000210958 0.00012845 0.000133952 0.00020485 0.000125923 C16:0 Palmitic acid 0.005358 0.0053779 0.0061470 0.0053097 0.0060347 0.0052760 0.0050159 0.0049306 0.0049889 0.0060344 0.002538354 0.003070414 0.003273197 0.002946034 0.003277811 0.001909507 0.002285865 0.002501848 0.001953368 C16:1 Palmitoleic acid 0.000041 0.0000622 0.0000398 0.0000521 0.0000395 0.0002100 0.0000752 0.0000572 0.0000543 0.0000447 6.81552E-05 0.000118109 8.13935E-05 9.92876E-05 9.64342E-05 8.76544E-05 9.60611E-05 0.000161769 8.22434E-05 C18:0 Stearic acid 0.007763 0.0087601 0.0098427 0.0083750 0.0097313 0.0096396 0.0089922 0.0091462 0.0090859 0.0114289 0.002946823 0.004116771 0.004835065 0.002714596 0.005504087 0.001211167 0.0018482 0.002472937 0.001754805 C18:1 Oleic acid 0.000990 0.0013580 0.0014745 0.0011874 0.0011753 0.0011291 0.0010816 0.0009032 0.0008759 0.0008354 0.001443836 0.002104352 0.002305743 0.001180631 0.002709462 0.00160119 0.002195088 0.002442533 0.002026323 C18:1 Vaccenic acid 0.000055 0.0000764 0.0000852 0.0000730 0.0000662 0.0000758 0.0000788 0.0000738 0.0000651 0.0000684 0.002108434 0.001772571 0.000336856 0.002251178 0.000386239 0.002611665 0.00314313 0.003769476 0.003117907 C18:2n-6 Linoleic acid 0.002104 0.0029148 0.0032520 0.0025534 0.0024719 0.0017675 0.0018880 0.0015190 0.0014448 0.0013550 0.001972418 0.00267421 0.002549921 0.00183258 0.002829984 0.001892408 0.002485484 0.002600439 0.002149047 C18:3 Linolenic acid 0.000989 0.0013561 0.0014707 0.0011804 0.0011722 0.0011428 0.0010691 0.0009064 0.0008614 0.0008257 0.000596123 0.000922014 0.000912282 0.000542379 0.001002392 0.000686754 0.000969541 0.001070388 0.000858384 C20:0 Arachidic acid 0.000042 0.0000449 0.0000379 0.0000288 0.0000472 0.0000288 0.0000339 0.0000340 0.0000475 0.0000553 NaN NaN NaN NaN NaN NaN NaN NaN NaN C20:1 Eicosenoic acid 0.000033 0.0000466 0.0000504 0.0000370 0.0000370 0.0000466 0.0000443 0.0000411 0.0000380 0.0000329 NaN NaN NaN NaN NaN NaN NaN NaN NaN C20:2 Eicosadienoic acid 0.000041 0.0000628 0.0000664 0.0000587 0.0000468 0.0000612 0.0000606 0.0000525 0.0000439 0.0000399 NaN NaN NaN NaN NaN NaN NaN NaN NaN C20:3 Dihomo-gamma-linolenic acid 0.000094 0.0001512 0.0001748 0.0001370 0.0001197 0.0001142 0.0001441 0.0001045 0.0000944 0.0000880 NaN NaN NaN NaN NaN NaN NaN NaN NaN C20:4n-6 Arachidonic acid 0.000370 0.0005546 0.0005970 0.0004690 0.0004552 0.0004547 0.0004891 0.0004055 0.0003558 0.0003076 9.83858E-05 0.000136977 0.000149478 9.43003E-05 0.000160195 0.000108133 0.000141513 0.000158948 0.000124987 C20:5n-6 Eicosapentanoic acid 0.000115 0.0001648 0.0001658 0.0001425 0.0001411 0.0001232 0.0001498 0.0001795 0.0001133 0.0001037 NaN NaN NaN NaN NaN NaN NaN NaN NaN C22:4 Adrenic acid 0.000076 0.0001202 0.0001221 0.0001123 0.0000991 0.0001022 0.0000994 0.0001019 0.0000900 0.0000776 NaN NaN NaN NaN NaN NaN NaN NaN NaN C22:5n-3 Docosapentanoic acid 0.000034 0.0000489 0.0000420 0.0000415 0.0000435 0.0000438 0.0000391 0.0000335 0.0000357 0.0000335 NaN NaN NaN NaN NaN NaN NaN NaN NaN C22:6 Docosahexaenoic acid 0.000056 0.0000868 0.0000804 0.0000679 0.0000704 0.0000590 0.0000663 0.0000504 0.0000553 0.0000528 NaN NaN NaN NaN NaN NaN NaN NaN NaN Cetyl alcohol 0.000048 0.0000497 0.0000573 0.0000540 0.0000580 0.0000631 0.0000643 0.0000600 0.0000599 0.0000716 NaN NaN NaN NaN NaN NaN NaN NaN NaN Cholesterol 0.144644 0.1217084 0.1254525 0.1188675 0.1644811 0.1311000 0.1203765 0.1392494 0.1498437 0.1839247 0.003317515 0.003378343 0.002857745 0.002536684 0.003009148 0.002072899 0.002474171 0.002178079 0.002264023 Citramalic acid 0.003386 0.0064561 0.0067097 0.0069535 0.0049329 0.0060847 0.0054428 0.0059941 0.0052555 0.0050560 0.007335345 0.008933417 0.008180756 0.008681317 0.007648709 0.006692897 0.007036948 0.0065924 0.007063715 Cysteine 0.000375 0.0000989 0.0000703 0.0000509 0.0001829 0.0000765 0.0000902 0.0000799 0.0002605 0.0001268 NaN NaN NaN NaN NaN NaN NaN NaN NaN Eryhtronic acid 0.000081 0.0000732 0.0000677 0.0000727 0.0000699 0.0002665 0.0002600 0.0003055 0.0002887 0.0001976 0.000168106 0.000213437 0.00018537 0.000196571 0.000186747 0.000304754 0.000340849 0.000373315 0.000306001 Ethanolamine 0.001459 0.0011318 0.0010640 0.0011700 0.0013366 0.0009047 0.0010524 0.0010966 0.0011278 0.0012053 0.002497884 0.002390672 0.002541148 0.005037611 0.002262721 0.003297381 0.002367701 0.002420762 0.002522511 Gluconic acid // Galactonic acid 0.000260 0.0002532 0.0002802 0.0002855 0.0002406 0.0005205 0.0005174 0.0005667 0.0005720 0.0004336 6.84554E-05 5.89526E-05 6.33049E-05 4.41417E-05 7.97669E-05 2.72471E-05 3.92142E-05 5.43769E-05 4.08526E-05 Glucose 0.267913 0.2922891 0.2687874 0.3430944 0.3174291 0.3868876 0.3944621 0.3910647 0.3475057 0.3438477 0.112748231 0.101799607 0.194233663 0.257002898 0.153812779 0.38003306 0.417957219 0.498582595 0.325010562 Glutaric acid 0.000169 0.0001555 0.0001466 0.0001773 0.0002015 0.0001553 0.0001540 0.0001648 0.0001678 0.0002376 6.1133E-05 7.86283E-05 6.31011E-05 4.92604E-05 4.8041E-05 5.20429E-05 4.25726E-05 3.70728E-05 5.04839E-05 Glyceric acid 0.000563 0.0005334 0.0005217 0.0005407 0.0005397 0.0003990 0.0004084 0.0004380 0.0004659 0.0004745 0.001949303 0.001973472 0.001895257 0.002038953 0.001984429 0.001128811 0.001089996 0.001275057 0.0011401 Glycerol 0.001818 0.0016209 0.0015793 0.0021099 0.0024493 0.0023274 0.0030155 0.0030803 0.0025781 0.0046185 0.003601601 0.004361327 0.003426591 0.004621806 0.003335315 0.009067165 0.00808062 0.008858499 0.007324436 Glycerol-1-phosphate 0.000120 0.0001493 0.0001307 0.0001395 0.0001251 0.0001106 0.0001074 0.0000986 0.0001106 0.0000807 0.000525056 0.000645257 0.000575732 0.000403929 0.000766 7.87186E-06 1.86711E-05 2.32166E-05 1.75472E-05 Hexose (Fructose) 1 0.001091 0.0010562 0.0008903 0.0010732 0.0010801 0.0013765 0.0013917 0.0014496 0.0014067 0.0006180 0.001457658 0.001325305 0.003081043 0.004108227 0.002422254 0.002434028 0.003073875 0.003798514 0.001993126 Hexose (Fructose) 2 0.000789 0.0007570 0.0006597 0.0008482 0.0008575 0.0011139 0.0011581 0.0011508 0.0010709 0.0005018 0.002257656 0.002041317 0.004833446 0.00570947 0.003914712 0.003696045 0.00438306 0.005544856 0.002919028 Hexose (Galactose) 0.000052 0.0000498 0.0000421 0.0000535 0.0000611 0.0000530 0.0000350 0.0000510 0.0000612 0.0003802 0.001026083 0.000939891 0.001465114 0.001771432 0.001261002 0.000408785 0.000422851 0.000489142 0.000304464 Hexose (L-Fucose // 6-Deoxyglucose) 1 0.000022 0.0000210 0.0000182 0.0000196 0.0000197 0.0000180 0.0000178 0.0000154 0.0000165 0.0000060 9.37791E-05 8.5029E-05 0.000103428 9.54399E-05 0.000107162 6.51302E-05 6.99919E-05 7.00729E-05 6.82787E-05 Hexose (L-Fucose // 6-Deoxyglucose) 2 0.000030 0.0000276 0.0000267 0.0000277 0.0000235 0.0000261 0.0000235 0.0000194 0.0000226 0.0000090 0.000116234 0.000101106 0.000136865 0.00012877 0.000141751 8.74245E-05 9.754E-05 9.63209E-05 8.69016E-05 Hexose 1 0.000313 0.0003353 0.0003124 0.0003016 0.0003676 0.0002596 0.0002640 0.0003271 0.0003318 0.0003286 5.53959E-05 5.91661E-05 0.000104499 8.22397E-05 9.73238E-05 0.000145348 0.000235581 0.000253721 0.000172205 Hexose 2 0.000567 0.0007135 0.0006550 0.0006048 0.0007589 0.0015560 0.0012935 0.0011224 0.0008152 0.0011058 0.001007343 0.000848636 0.001493642 0.001407937 0.001182431 0.002576355 0.004789106 0.005732792 0.002885184 Oxalic acid 0.000209 0.0002000 0.0002511 0.0002108 0.0002308 0.0002489 0.0002376 0.0002450 0.0002221 0.0002449 0.00018615 0.000328573 0.000412153 2.57647E-05 0.000290793 0.000164693 0.000334369 0.000289804 0.000235843 Phosphodimethylethanolamine 0.023855 0.0230529 0.0229863 0.0203746 0.0228681 0.0147607 0.0146970 0.0156161 0.0199755 0.0162965 0.001003276 0.001110456 0.001081791 0.000731976 0.00145599 0.000178586 0.000485188 0.001136531 0.000582463 Phosphoric acid 0.026783 0.0269462 0.0269404 0.0292497 0.0225800 0.0264049 0.0218966 0.0211291 0.0228247 0.0153033 0.002765063 0.002416827 0.002102263 0.002510288 0.002502161 0.001428265 0.002021481 0.002570833 0.001948881 Picolinic acid 0.000286 0.0003853 0.0004349 0.0004939 0.0004225 0.0005559 0.0005147 0.0005883 0.0004025 0.0005026 0.000689945 0.000635804 0.000697713 0.000616953 0.000607457 0.00083984 0.000684611 0.000494181 0.000523648 Sorbitol 0.000100 0.0000701 0.0000447 0.0000814 0.0000858 0.0005682 0.0005557 0.0006004 0.0005874 0.0007777 0.000107881 0.000126834 0.000118571 0.000135048 0.0001979 0.000181624 0.000163706 0.000208994 0.000143939 Threonic acid 0.000037 0.0000342 0.0000328 0.0000309 0.0000341 0.0002149 0.0002139 0.0002183 0.0002179 0.0001512 0.00011424 0.000126508 0.000111321 0.00011309 0.000123559 0.000100984 0.000115347 0.000131108 0.000125667 Uracil 0.000245 0.0002220 0.0002294 0.0002313 0.0002512 0.0002019 0.0001795 0.0002057 0.0001947 0.0001885 0.000848038 0.00089797 0.001012892 0.000694605 0.001052533 0.000126366 7.70662E-05 7.34833E-05 5.8648E-05 Urea 0.000164 0.0000341 0.0000141 0.0000760 0.0000820 0.0000953 0.0001491 0.0000984 0.0000795 0.0001128 0.036795337 0.040799422 0.058610448 0.030901472 0.05791094 0.036540595 0.045662768 0.039609109 0.038616601 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG COMPOUND ID feature (m/z@RT) calculated M RT ppm error ΔRT error 1-Methylhydantoin // Dihidrouracil C02565 // C00429 259.1294@10.018 259.1294 10.018 -5.4 -3.47 2-Hydroxybutyric acid C05984 249.1337@7.825 249.1337 7.825 -2.01 -2.79 2-Hydroxyglutaric acid C02630 349.1311@11.603 349.1311 11.603 -6.3 - Alanine C00041 234.1273@7.663 234.1273 7.663 1.28 -0.56 Aminomalonic acid C00872 320.1216@10.657 320.1216 10.657 -6.25 - Arabitol // Ribitol C00532 // C00474 243.1171@13.696 243.1171 13.696 0 -2.54 beta-Glycerophosphate C02979 445.1474@13.394 445.1474 13.394 -4.34 -0.74 C14:0 Myristic acid C06424 301.2542@14.853 301.2542 14.853 4.98 -1.14 C15:0 Pentadecanoic acid C16537 315.2684@16.257 315.2684 16.257 -6.66 - C16:0 Palmitic acid C00249 313.2462@17.817 313.2462 17.817 1.21 -1.68 C16:1 Palmitoleic acid C08362 325.2619@17.424 325.2619 17.424 -5.81 -1.04 C18:0 Stearic acid C01530 357.3182@21.279 357.3182 21.279 -1.4 -1.38 C18:1 Oleic acid C00712 356.2953@20.791 356.2953 20.791 -2.53 -1.1 C18:1 Vaccenic acid C08367 356.3055@20.911 356.3055 20.911 5.07 - C18:2n-6 Linoleic acid C01595 353.2858@20.665 353.2858 20.665 -3.4 -1.32 C18:3 Linolenic acid C06427 356.2953@20.776 356.2953 20.776 -3.7 - C20:0 Arachidic acid C06425 385.3496@25.159 385.3496 25.159 -8.04 -2 C20:1 Eicosenoic acid C16526 383.3340@24.630 383.334 24.63 -1.57 - C20:2 Eicosadienoic acid C16525 381.3183@24.514 381.3183 24.514 -4.72 - C20:3 Dihomo-gamma-linolenic acid C03242 379.3079@24.096 379.3079 24.096 -2.37 - C20:4n-6 Arachidonic acid C00219 378.2888@23.674 378.2888 23.674 -5.57 - C20:5n-6 Eicosapentanoic acid C06428 375.2714@23.785 375.2714 23.785 -4 - C22:4 Adrenic acid C16527 406.3127@27.291 406.3127 27.291 -0.99 - C22:5n-3 Docosapentanoic acid C16513 403.3027@27.367 403.3027 27.367 10.91 - C22:6 Docosahexaenoic acid C06429 401.2855@27.125 401.2855 27.125 -3.74 - Cetyl alcohol C00823 313.2935@16.549 313.2935 16.549 -10.47 -1.87 Cholesterol C00187 369.3423@26.245 369.3423 26.245 -6.53 15.07 Citramalic acid C02614 276.1345@10.712 276.1345 10.712 7.39 -9.22 Cysteine C00097 Cysteine, 3TMS Cysteine, 3TMS Cysteine, 3TMS -6.51 0.67 Eryhtronic acid NA 409.1727@11.467 409.1727 11.467 0.24 - Ethanolamine C00189 262.1413@9.042 262.1413 9.042 8.27 - Gluconic acid // Galactonic acid C00257 // C00880 467.2119@15.638 467.2119 15.638 -1.93 -2.3 Glucose C00031 568.267@17.072 + 568.267@17.545 568.267 17.545 -0.51 - Glutaric acid C00489 277.1281@11.034 277.1281 11.034 0.72 - Glyceric acid C00258 307.12@9.414 307.12 9.414 -4.02 -0.32 Glycerol C00116 293.1402@8.976 293.1402 8.976 -11 -1.3 Glycerol-1-phosphate C00623 448.137@13.837 448.137 13.837 0 -0.81 Hexose (Fructose) 1 C00095 568.267@16.403 568.267 16.403 -8.21 - Hexose (Fructose) 2 C00095 568.267@16.453 568.267 16.453 -8.73 - Hexose (Galactose) C00095 568.267@16.771 568.267 16.771 -17.29 - Hexose (L-Fucose // 6-Deoxyglucose) 1 C01019 // NA 480.2417@14.854 480.2417 14.854 -0.6 - Hexose (L-Fucose // 6-Deoxyglucose) 2 C01019 // NA 480.2417@15.07 480.2417 15.07 -9.87 - Hexose 1 C00738 188.1045@16.121 188.1045 16.121 1.56 - Hexose 2 C00738 226.2157@16.131 226.2157 16.131 1.56 - Oxalic acid C00209 235.0816@7.699 235.0816 7.699 0 0.58 Phosphodimethylethanolamine C13482 314.1374@10.541 314.1374 10.541 2.23 - Phosphoric acid C00009 613.1518@8.951 613.1518 8.951 5.08 -0.64 Picolinic acid C10164 196.0788@9.219 196.0788 9.219 -4.59 1.28 Sorbitol C00794 345.1708@17.138 345.1708 17.138 -8.45 -4.72 Threonic acid C01620 409.1727@11.588 409.1727 11.588 -0.71 - Uracil C00106 257.1062@9.434 257.1062 9.434 -19.06 -0.22 Urea C00086 205.1187@8.51 205.1187 8.51 -2.93 -3.47 METABOLITES_END #END