#METABOLOMICS WORKBENCH Zhou999_20161213_161728 DATATRACK_ID:798 STUDY_ID:ST000522 ANALYSIS_ID:AN000797 PROJECT_ID:PR000385 VERSION 1 CREATED_ON December 19, 2016, 10:24 am #PROJECT PR:PROJECT_TITLE Metabolomics profiling studies for snout bream fish PR:PROJECT_TYPE LC-MS analysis PR:PROJECT_SUMMARY Metabolomics profiling studies on plasma samples from three different blunt PR:PROJECT_SUMMARY snout bream groups PR:INSTITUTE Huazhong Agricultural University, Wuhan PR:DEPARTMENT College of Fisheries PR:LABORATORY Key Lab of Freshwater Animal Breeding, Ministry of Agriculture PR:LAST_NAME Zhou PR:FIRST_NAME LaiFang PR:ADDRESS Hongshan, Wuhan, 430070 Hubei, China PR:EMAIL 156851836@qq.com PR:PHONE 15171617087 #STUDY ST:STUDY_TITLE Plasma metabolomics profiling for fish maturation in blunt snout bream ST:STUDY_TYPE Metabolomics experiment ST:STUDY_SUMMARY We investigated the comprehensive metabolic profiles of plasma among immature ST:STUDY_SUMMARY females, mature females ready to spawn, as well as already spawned breeders of ST:STUDY_SUMMARY blunt snout bream (Megalobrama amblycephala). The purpose of this study was to ST:STUDY_SUMMARY screen out potential biomarkers for sexual mature female M. amblycephala ST:STUDY_SUMMARY compared to immature female individuals and already spawned breeders. The three ST:STUDY_SUMMARY groups were set up in this study, including one year old immature females, 2 ST:STUDY_SUMMARY years old sexually mature females ready to spawning and successfully spawning ST:STUDY_SUMMARY females of M. amblycephala. The plasma samples were collected to investigate the ST:STUDY_SUMMARY comprehensive metabolic profiles through UPLC-MS/MS based metabolomics analysis ST:STUDY_SUMMARY method. According to multivariate and univariate statistical analysis, plasma ST:STUDY_SUMMARY metabolite profiles of three groups were obviously separated, and the plasma ST:STUDY_SUMMARY metabolite profiles of immature female M. amblycephala were much more different ST:STUDY_SUMMARY from mature females ready to spawn as well as already spawned breeders. The ST:STUDY_SUMMARY differential plasma metabolites from three hormone related pathways including ST:STUDY_SUMMARY GnRH signaling pathway, steroid hormone biosynthesis and steroid biosynthesis, ST:STUDY_SUMMARY were further analyzed. A total of 29 metabolites were identified as differential ST:STUDY_SUMMARY biomarkers associated with the female maturation status ST:INSTITUTE Huazhong Agricultural University, Wuhan ST:DEPARTMENT College of Fisheries ST:LABORATORY Key Lab of Freshwater Animal Breeding, Ministry of Agriculture ST:LAST_NAME Zhou ST:FIRST_NAME LaiFang ST:ADDRESS Hongshan, Wuhan, 430070 Hubei, China ST:EMAIL 156851836@qq.com ST:PHONE 15171617087 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Megalobrama amblycephala SU:TAXONOMY_ID 75352 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS P1503530001 20150724_YU_2_1_1 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530002 20150724_YU_2_1_2 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530003 20150724_YU_2_1_3 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530004 20150724_YU_2_1_4 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530005 20150724_YU_2_1_5 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530006 20150724_YU_2_1_6 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530007 20150724_YU_2_1_7 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530008 20150724_YU_2_1_8 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530009 20150724_YU_2_1_9 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530010 20150724_YU_2_1_10 Phenotype:2 years old sexually mature without hormone treatment gender=female SUBJECT_SAMPLE_FACTORS P1503530011 20150724_YU_2_2_1 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530012 20150724_YU_2_2_2 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530013 20150724_YU_2_2_3 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530014 20150724_YU_2_2_4 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530015 20150724_YU_2_2_5 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530016 20150724_YU_2_2_6 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530017 20150724_YU_2_2_11 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530018 20150724_YU_2_2_8 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530019 20150724_YU_2_2_9 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530020 20150724_YU_2_2_10 Phenotype:2 years old sexually mature after successfully spawning gender=female SUBJECT_SAMPLE_FACTORS P1503530021 20150724_YU_1_1_1 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530022 20150724_YU_1_1_3 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530023 20150724_YU_1_1_5 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530024 20150724_YU_1_1_6 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530025 20150724_YU_1_1_9 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530026 20150724_YU_1_1_10 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530027 20150724_YU_1_1_11 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530028 20150724_YU_1_1_12 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530029 20150724_YU_1_1_14 Phenotype:1 year old immature gender=female SUBJECT_SAMPLE_FACTORS P1503530030 20150724_YU_1_1_22 Phenotype:1 year old immature gender=female #COLLECTION CO:COLLECTION_SUMMARY About 2 ml of blood was collected via caudal vein by syringe for each female CO:COLLECTION_SUMMARY M.amblycephala and then directly put into K3 EDTA centrifuge tube. CO:COLLECTION_PROTOCOL_FILENAME Zhou999_20161213_161728_PR_CO_collection_files.docx CO:SAMPLE_TYPE Plasma #TREATMENT TR:TREATMENT_SUMMARY The blood was centrifuged at 4000 rpm for 5 min at 4°C as soon as it was TR:TREATMENT_SUMMARY extracted from fish and about 1 ml supernatant was pipetted into another TR:TREATMENT_SUMMARY tube.The extracted plasma samples were then frozen in liquid nitrogen TR:TREATMENT_SUMMARY immediately and stored in the -80°C. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To initiate the experiment, each plasma sample was mixed with the same volume SP:SAMPLEPREP_SUMMARY and labeled as Quality control (QC) sample. Plasma sample (100 µl, including QC SP:SAMPLEPREP_SUMMARY sample) was added into new Eppendorf tube with ice-cold methanol (300 µl), SP:SAMPLEPREP_SUMMARY vortex mixing for 1 min, standing for 1 min, and make sure the solution volume SP:SAMPLEPREP_SUMMARY had no obvious observable difference. Then samples were precipitated for 20 min SP:SAMPLEPREP_SUMMARY at -20°C, and centrifuged at 14000 rpm for 20 min at 4°C to precipitate the SP:SAMPLEPREP_SUMMARY proteins. A total of 250 µl of protein free supernatant was collected and dried SP:SAMPLEPREP_SUMMARY with nitrogen at 37°C. The concentration of extractant was diluted depending on SP:SAMPLEPREP_SUMMARY the mass spectrum response signals. SP:SAMPLEPREP_PROTOCOL_FILENAME Zhou999_20161213_161728_PR_SP_sampleprep.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Chromatographic separation was performed on a ACQUITY UPLC BEH C18 analytical CH:CHROMATOGRAPHY_SUMMARY column (100×2.1 mm, 1.7 µm, Waters, Milford, USA) using a 2777C UPLC system CH:CHROMATOGRAPHY_SUMMARY (Waters). The flow rate was 0.40 mL/min and the mobile phase was composed of CH:CHROMATOGRAPHY_SUMMARY solvents A (95% H2O/5% acetonitrile + 0.1% formic acid) and B (95% CH:CHROMATOGRAPHY_SUMMARY acetonitrile/5% H2O＋0.1% formic acid).The gradient program was optimized as CH:CHROMATOGRAPHY_SUMMARY follows: 0–0.1 min, 100% A; 0.1–0.6 min, 100% A-50% A; 0.6–5 min, 50%–0% CH:CHROMATOGRAPHY_SUMMARY A; 5–8 min, 0% A; 8–10 min, 0% A-100% A. Injection volume was 10 µl. The CH:CHROMATOGRAPHY_SUMMARY eluent from the column was directly added in to the mass spectrometer without CH:CHROMATOGRAPHY_SUMMARY split. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters 2777C CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2mm, 1.7um) CH:FLOW_GRADIENT 100% acetonitrile CH:FLOW_RATE 0.40 mL/min CH:SOLVENT_A 95% H2O/5% acetonitrile + 0.1% formic acid CH:SOLVENT_B 95% acetonitrile/5% H2O＋0.1% formic acid #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Waters Synapt G2 XS QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak intensity MS_METABOLITE_DATA_START Samples 20150724_YU_2_1_1 20150724_YU_2_1_2 20150724_YU_2_1_3 20150724_YU_2_1_4 20150724_YU_2_1_5 20150724_YU_2_1_6 20150724_YU_2_1_7 20150724_YU_2_1_8 20150724_YU_2_1_9 20150724_YU_2_1_10 20150724_YU_1_1_1 20150724_YU_1_1_3 20150724_YU_1_1_5 20150724_YU_1_1_6 20150724_YU_1_1_10 20150724_YU_1_1_11 20150724_YU_1_1_12 20150724_YU_1_1_14 20150724_YU_1_1_22 20150724_YU_2_2_1 20150724_YU_2_2_2 20150724_YU_2_2_3 20150724_YU_2_2_4 20150724_YU_2_2_5 20150724_YU_2_2_6 20150724_YU_2_2_11 20150724_YU_2_2_8 20150724_YU_2_2_9 20150724_YU_2_2_10 20150724_YU_2_1_1 20150724_YU_2_1_2 20150724_YU_2_1_3 20150724_YU_2_1_4 20150724_YU_2_1_5 20150724_YU_2_1_6 20150724_YU_2_1_7 20150724_YU_2_1_8 20150724_YU_2_1_9 20150724_YU_2_1_10 20150724_YU_1_1_1 20150724_YU_1_1_3 20150724_YU_1_1_5 20150724_YU_1_1_6 20150724_YU_1_1_10 20150724_YU_1_1_11 20150724_YU_1_1_12 20150724_YU_1_1_14 20150724_YU_1_1_22 20150724_YU_2_2_1 20150724_YU_2_2_2 20150724_YU_2_2_3 20150724_YU_2_2_4 20150724_YU_2_2_5 20150724_YU_2_2_6 20150724_YU_2_2_11 20150724_YU_2_2_8 20150724_YU_2_2_9 20150724_YU_2_2_10 Factors Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:2 years old sexually mature without hormone treatment Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:1 year old immature Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Phenotype:2 years old sexually mature after successfully spawning Arachidonic acid 5747478.363 16158753.25 34671966.14 6893354.446 30953796.66 5455686.715 25273193.37 50198787.47 33544065.6 5841034 4439578.012 3607384.831 6537205.543 12849230.51 1616053.029 1633770.465 1965856.826 2005117.63 15275367.89 4815588.026 16354333.25 23295863.81 4698466.991 16706036.05 3827491.199 8512661.297 45774486.56 12522144.06 3716631.972 7523746.354 23565823.41 41911623.04 7852764.154 39294814.47 5019241.148 27726491.11 56792303.46 47075265.48 8284981.321 3371212.313 2000878.341 5965113.827 12962714.17 1455682.557 1192113.755 1355709.436 1570691.949 16996488.11 5063294.162 21302274.63 24860320.91 4196012.182 16157518.8 2964570.555 8157857.514 52504349.74 12147604.82 3683813.586 Testosterone glucuronide 47709.81105 45586.35527 36284.12679 59351.08894 49031.42559 57562.22729 47233.42279 64332.55992 32543.96136 34333.70313 78855.57847 92095.1624 93895.0992 114093.3965 98575.08778 93560.27221 96119.54275 71763.97149 55518.68727 56733.89225 46465.8663 33168.94147 60351.28282 72480.08807 92008.64618 19052.82512 34308.28824 58962.31678 56325.72072 62454.60953 66482.85184 43860.40983 67611.5101 62243.76262 52957.34797 51818.42507 72782.52025 45671.73339 48699.26952 59879.31654 51081.66287 85678.03947 115101.0627 88792.90057 68268.15017 66286.70479 56215.70055 61774.13958 59652.19281 60523.93757 35396.43501 53897.30702 70100.31477 71264.99034 18258.71219 39352.36635 57198.74489 55828.35665 17a,21-Dihydroxypreg-nenolone 1714.24499 4790.785758 4282.871478 3518.273645 3957.938291 8035.55824 2087.222418 2568.542069 1616.063476 3330.099259 7645.620739 13874.11146 6537.576278 19349.87833 11249.99091 11741.76477 4122.608711 9854.834586 8813.194834 7508.15451 5054.013251 6655.150933 7540.48892 4771.148607 5377.543225 3631.375756 6941.317295 4881.688178 8213.451785 2244.035328 6986.85161 5177.153618 4007.943212 5024.470907 7392.727382 2289.831482 2905.915222 2267.960541 4723.446252 5805.734398 7695.438782 5965.452118 19520.77533 10133.58798 8567.616807 2843.065404 7719.701385 9806.203183 7894.361954 6583.085754 7102.084269 6734.107833 4614.495209 4165.158188 3480.021695 7961.84465 4735.676139 8140.925845 3a,21-Dihydroxy-5b-pregnane-11,20-dione 1714.24499 4790.785758 4282.871478 3518.273645 3957.938291 8035.55824 2087.222418 2568.542069 1616.063476 3330.099259 7645.620739 13874.11146 6537.576278 19349.87833 11249.99091 11741.76477 4122.608711 9854.834586 8813.194834 7508.15451 5054.013251 6655.150933 7540.48892 4771.148607 5377.543225 3631.375756 6941.317295 4881.688178 8213.451785 2244.035328 6986.85161 5177.153618 4007.943212 5024.470907 7392.727382 2289.831482 2905.915222 2267.960541 4723.446252 5805.734398 7695.438782 5965.452118 19520.77533 10133.58798 8567.616807 2843.065404 7719.701385 9806.203183 7894.361954 6583.085754 7102.084269 6734.107833 4614.495209 4165.158188 3480.021695 7961.84465 4735.676139 8140.925845 Androsterone 6793.131954 2428.73884 16924.73435 23678.79514 26447.96316 13275.52343 11277.40572 6424.862649 5045.997012 3431.591373 4174.714602 6382.723704 1236.580581 9534.588037 0 0 0 0 0 13729.03188 2584.724264 10460.47039 7299.384191 20324.76186 4424.613275 17535.15683 16357.96496 2881.50095 792.1118914 8892.560972 3542.057344 20458.69229 26974.38456 33574.80883 12213.50436 12372.11637 7268.756233 7081.480575 4867.403687 3170.087164 3540.252625 1128.363468 9618.796966 0 0 0 0 0 14435.23129 3366.722767 11162.95378 6518.786883 19657.42926 3427.06947 16804.29961 18762.94805 2795.314817 785.1174315 5b-Dihydrotestosterone 6793.131954 2428.73884 16924.73435 23678.79514 26447.96316 13275.52343 11277.40572 6424.862649 5045.997012 3431.591373 4174.714602 6382.723704 1236.580581 9534.588037 0 0 0 0 0 13729.03188 2584.724264 10460.47039 7299.384191 20324.76186 4424.613275 17535.15683 16357.96496 2881.50095 792.1118914 8892.560972 3542.057344 20458.69229 26974.38456 33574.80883 12213.50436 12372.11637 7268.756233 7081.480575 4867.403687 3170.087164 3540.252625 1128.363468 9618.796966 0 0 0 0 0 14435.23129 3366.722767 11162.95378 6518.786883 19657.42926 3427.06947 16804.29961 18762.94805 2795.314817 785.1174315 Etiocholanolone 6793.131954 2428.73884 16924.73435 23678.79514 26447.96316 13275.52343 11277.40572 6424.862649 5045.997012 3431.591373 4174.714602 6382.723704 1236.580581 9534.588037 0 0 0 0 0 13729.03188 2584.724264 10460.47039 7299.384191 20324.76186 4424.613275 17535.15683 16357.96496 2881.50095 792.1118914 8892.560972 3542.057344 20458.69229 26974.38456 33574.80883 12213.50436 12372.11637 7268.756233 7081.480575 4867.403687 3170.087164 3540.252625 1128.363468 9618.796966 0 0 0 0 0 14435.23129 3366.722767 11162.95378 6518.786883 19657.42926 3427.06947 16804.29961 18762.94805 2795.314817 785.1174315 Dihydrotestosterone 6793.131954 2428.73884 16924.73435 23678.79514 26447.96316 13275.52343 11277.40572 6424.862649 5045.997012 3431.591373 4174.714602 6382.723704 1236.580581 9534.588037 0 0 0 0 0 13729.03188 2584.724264 10460.47039 7299.384191 20324.76186 4424.613275 17535.15683 16357.96496 2881.50095 792.1118914 8892.560972 3542.057344 20458.69229 26974.38456 33574.80883 12213.50436 12372.11637 7268.756233 7081.480575 4867.403687 3170.087164 3540.252625 1128.363468 9618.796966 0 0 0 0 0 14435.23129 3366.722767 11162.95378 6518.786883 19657.42926 3427.06947 16804.29961 18762.94805 2795.314817 785.1174315 4a-Methylzymosterol-4-carboxylic acid 57094.34862 40488.09785 39861.22769 24500.43246 52218.60269 67377.5484 50492.75552 55352.48786 45614.95492 57321.21895 48120.69864 95265.37066 57375.79028 76831.90759 97034.82082 71857.58964 85618.09927 58041.59851 57835.20699 61226.93429 24756.43237 17720.13666 15805.77425 43859.23659 41567.60473 21621.33509 31822.7812 27708.3404 45466.0319 74739.45445 59047.5855 48184.42491 27910.37648 66289.77785 61987.46025 55394.14495 62622.93267 64015.3802 81304.99296 36540.65574 52840.05609 52354.65179 77510.48254 87405.48338 52432.34767 59044.61786 45466.39566 64351.66833 64376.34973 32246.39691 18910.15021 14115.50223 42419.18536 32196.04975 20720.16785 36501.43474 26879.57971 45064.56042 4a-Carboxy-4b-methyl-5a-cholesta-8,24-dien-3b-ol 57094.34862 40488.09785 39861.22769 24500.43246 52218.60269 67377.5484 50492.75552 55352.48786 45614.95492 57321.21895 48120.69864 95265.37066 57375.79028 76831.90759 97034.82082 71857.58964 85618.09927 58041.59851 57835.20699 61226.93429 24756.43237 17720.13666 15805.77425 43859.23659 41567.60473 21621.33509 31822.7812 27708.3404 45466.0319 74739.45445 59047.5855 48184.42491 27910.37648 66289.77785 61987.46025 55394.14495 62622.93267 64015.3802 81304.99296 36540.65574 52840.05609 52354.65179 77510.48254 87405.48338 52432.34767 59044.61786 45466.39566 64351.66833 64376.34973 32246.39691 18910.15021 14115.50223 42419.18536 32196.04975 20720.16785 36501.43474 26879.57971 45064.56042 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Arachidonic acid 3.48 303.2325 444899 C00219 Testosterone glucuronide 1.07 463.2336 108192 C11134 17a,21-Dihydroxypreg-nenolone 1.97 385.1783 192735 C05487 3a,21-Dihydroxy-5b-pregnane-11,20-dione 1.97 385.1783 44263347 C05478 Androsterone 3.04 289.2166 12306765 C00523 5b-Dihydrotestosterone 3.04 289.2166 11302 C05293 Etiocholanolone 3.04 289.2166 5880 C04373 Dihydrotestosterone 3.04 289.2166 10635 C03917 4a-Methylzymosterol-4-carboxylic acid 4.52 441.3365 53477909 C15808 4a-Carboxy-4b-methyl-5a-cholesta-8,24-dien-3b-ol 4.52 441.3365 23724596 C15808 METABOLITES_END #END