#METABOLOMICS WORKBENCH dagaul_20171214_102338 DATATRACK_ID:1284 STUDY_ID:ST000922 ANALYSIS_ID:AN001512 PROJECT_ID:PR000638 VERSION 1 CREATED_ON January 22, 2018, 5:17 pm #PROJECT PR:PROJECT_TITLE Metabolites associated with risk perception and predator detection PR:PROJECT_SUMMARY Blue Crab fed either mud crab or oyster diet to investigator behavior changes of PR:PROJECT_SUMMARY mud crabs due to compounds released in blue crab urine. PR:INSTITUTE Georgia Institute of Technology PR:DEPARTMENT School of Chemistry and Biochemistry PR:LABORATORY Systems Mass Spectrometry Core (SyMSC) PR:LAST_NAME Gaul PR:FIRST_NAME David PR:ADDRESS 311 Ferst Drive, Atlanta, GA, 30332, USA PR:EMAIL david.gaul@chemistry.gatech.edu PR:PHONE 4043850377 PR:FUNDING_SOURCE NSF #STUDY ST:STUDY_TITLE Crab Urine Study ST:STUDY_SUMMARY Metabolites from blue crab urine associated with different diets ST:INSTITUTE Georgia Institute of Technology ST:DEPARTMENT School of Chemistry and Biochemistry ST:LABORATORY SyMSC ST:LAST_NAME Gaul ST:FIRST_NAME David ST:ADDRESS 311 Ferst Drive, Atlanta, GA, 30332, USA ST:EMAIL david.gaul@chemistry.gatech.edu ST:PHONE 4043850377 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 8 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Callinectes sapidus SU:TAXONOMY_ID 6763 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - MA Diet:mud crab diet SUBJECT_SAMPLE_FACTORS - MB Diet:mud crab diet SUBJECT_SAMPLE_FACTORS - MC Diet:mud crab diet SUBJECT_SAMPLE_FACTORS - MD Diet:mud crab diet SUBJECT_SAMPLE_FACTORS - OA Diet:oyster diet SUBJECT_SAMPLE_FACTORS - OB Diet:oyster diet SUBJECT_SAMPLE_FACTORS - OC Diet:oyster diet SUBJECT_SAMPLE_FACTORS - OD Diet:oyster diet #COLLECTION CO:COLLECTION_SUMMARY Urine was collected from Blue Crabs CO:SAMPLE_TYPE Urine CO:COLLECTION_METHOD 22-gauge needle with gentle suction CO:COLLECTION_LOCATION Skidway Institute of Oceanography CO:COLLECTION_FREQUENCY every other day CO:STORAGE_CONDITIONS -20C #TREATMENT TR:TREATMENT_SUMMARY Blue crabs were fed mud crabs or oysters #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Dilute and shoot, urine SP:PROCESSING_METHOD mixed 1 (sample): 5 (MeOH), centrifuged, supernatent analyzed SP:PROCESSING_STORAGE_CONDITIONS on ice #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Basic pH HILIC method CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters Acquity BEH Amide (75 x 2.1mm, 1.7um) CH:FLOW_GRADIENT (time, %A): 0, 5; 3, 63; 7, 63; 7.1, 5; 7.2, 0.5; 9.5, 5; 9.9,5. CH:FLOW_RATE (time, rate (ml/min)): 0, 0.3; 3, 0.3; 7, 0.3; 7.1, 0.3; 7.2, 0.5; 9.5, 0.5; CH:FLOW_RATE 9.9, 0.3. CH:COLUMN_TEMPERATURE 50 CH:SOLVENT_A 10mM NaOAc, 0,05% NH4OH CH:SOLVENT_B ACN, 0.05% NH4OH CH:SAMPLE_INJECTION 2 ul #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME SyMSC AN:OPERATOR_NAME David Gaul AN:SOFTWARE_VERSION Xcalibur 4.0 AN:DATA_FORMAT profile #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:CAPILLARY_TEMPERATURE 275 C MS:COLLISION_ENERGY 10, 30, 50 NCE stepped MS:COLLISION_GAS nitrogen MS:FRAGMENTATION_METHOD HCD MS:SPRAY_VOLTAGE 2.80 kV MS:AUTOMATIC_GAIN_CONTROL 3.00E+6 MS:RESOLUTION_SETTING 240,000 MS:SCANNING_RANGE 70-1050 MS:MS_RESULTS_FILE ST000922_AN001512_Results.txt UNITS:peak area #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples MA MB MC MD OA OB OC OD Factors Diet:mud crab diet Diet:mud crab diet Diet:mud crab diet Diet:mud crab diet Diet:oyster diet Diet:oyster diet Diet:oyster diet Diet:oyster diet [FAamino_oxo(6:0/2:0)]2-amino-3-oxo-hexanedioicacid 155294.9104 101852.1661 37573.02566 70063.91251 63877.3774 51145.089 2,5-dichlorobenzoquinone 199413.1634 291568.1953 223530.8738 390404.1067 257133.6184 225966.3968 2_4-Dinitrophenylhydrazine 782572.1943 1613751.713 2653518.488 559727.5388 3052887.239 633949.9287 2-Amino-4-hydroxypteridine-6-carbaldehyde 1291688.601 1132064.077 876000.1663 333631.8828 47228.31361 707446.7889 2-oxo-1_2-dihydroquinoline-4-carboxylate_R1 43223.56595 232429.4753 100191.6418 9252.006364 145574.8905 34828.59418 2-oxo-1_2-dihydroquinoline-4-carboxylate_R2 153926.1496 465972.7059 95100.82563 21586.76734 23057.2155 68306.57352 3-hydroxy-1H-quinolin-4-one_R1 184532.3415 513170.8832 247246.5148 5697.313331 170126.6395 112315.2245 3-hydroxy-1H-quinolin-4-one_R2 425479.3121 615520.1715 124822.584 5556.232053 248583.7459 82404.29554 3-Sulfamoylalanine 9125.910689 37266.43551 47718.22273 63454.84254 417787.9628 61296.36043 4-Formyl-2-methoxyphenyl hydrogen sulfate 122059.4395 321751.2792 207867.4791 24073.08537 146588.8041 25772.6567 4-Hydroxy-8-(beta-D-talopyranosyloxy)-2-quinolinecarboxylic acid 62142.23404 159335.8058 124632.5694 71252.0261 734349.6232 374238.5634 4-Methylene-L-glutamine 250114.8865 596449.7315 462523.1981 78559.26652 499983.2164 359826.7596 5-Hydroxyisourate 3983.27283 8063.397648 216396.4434 14094.16144 479040.6748 11132.75414 5-L-Glutamyl-taurine 150864.2231 238452.4131 83268.84221 33881.49069 42206.18296 149112.305 6-Acetyl-2-amino-1,7,8,9-tetrahydro-4H-pyrimido[4,5-b][1,4]diazepin-4-one 323779.5796 612337.3332 174560.9462 14926.0673 5354.744651 235590.8656 6-Aminopenicillanate_R1 1530.790814 2206.343759 4400.793143 195012.4977 20492.08798 6609.752004 6-Aminopenicillanate_R2 33549.68816 34149.52492 10589.99866 598857.9716 43256.44245 33059.44832 Acamprosate 162283.172 275930.2837 140863.9246 18715.0875 148536.4907 85802.57547 Asulam 1489718.392 2363835.994 979789.0945 7676130.102 1445651.937 1129821.738 beta-D-glucuronamide 55189.12103 397093.3012 68384.77635 12387.34003 2012.508806 28224.56577 capuride 9042.776067 73544.5142 1430.349831 4645.768207 3737.441286 5277.203688 Cholinesulfate_R1 160093.0406 390902.3618 282640.1383 120998.9801 173137.8428 216736.9415 Cholinesulfate_R2 138273.5356 277168.3921 237616.2076 34853.8537 316267.5482 185430.2783 diethylmaleate_R1 238909.7443 393426.8457 364022.0162 261324.9273 214235.3809 370228.9684 diethylmaleate_R2 124493.567 180255.1525 364022.0162 261324.9273 214235.3809 370228.9684 DL-Homolanthionine sulfone 9426.646426 42878.68207 1537.021658 4840.374545 1633.45204 1625.383939 Ethyl N-acetyl-N-butyl-beta-alaninate 11250.78798 83060.74046 26869.44251 7896.206756 8485.675909 159044.3836 Hydroxyanthraquinone 428008.476 353859.6655 462275.8075 226188.2031 387867.7625 368731.5959 L-Homocysteicacid 14148.91512 133986.9071 194677.1915 8187.302577 181298.4695 264296.852 L-Threonicacid 1079536.845 816943.0715 991866.5381 629127.0328 1653364.827 386957.3102 nitecapone 21570.96089 83449.92295 23498.44003 1021.168957 116674.7258 6242.304709 N-Undecanoylglycine 524984.9916 1185502.093 167685.0028 186104.5464 209357.1127 2178345.683 Pantothenicacid(VitaminB5) 32169.98894 64316.43133 83247.98942 1825.355237 61024.26376 39257.0717 Sulfanilic acid 818230.0597 1241623.474 665840.2435 76857.22926 342213.5494 440692.2335 Sulfate 13306514.9 13533007.22 13423879.12 16474908.78 16855740.09 14445215.7 Sulfosalicylic Acid 32598.50339 642227.5028 43700.22947 105323.7214 126484.184 145336.589 Tauropine 178504.6739 311471.754 322700.0377 134463.8875 67825.12811 445055.0982 tyramine sulfate_R1 8990874.581 1674736.212 1082817.514 3286072.047 808637.9562 2254718.525 tyramine sulfate_R2 23539821.99 3192382.863 1978220.388 5636629.159 1237317.273 3771575.984 Xanthopterin-B2 1041351.733 1333485.572 566666.9363 46912.99577 6536.121573 373943.0997 Xanthurenicacid_R1 852268.4635 2290804.766 1271929.192 34560.25568 4050076.616 534427.4107 Xanthurenicacid_R2 274090.0101 1463174.234 189886.4949 7442.432519 968814.867 70806.03776 Xanthurenicacid_R3 1981920.765 3322177.379 628722.0012 35525.12005 1376521.221 420848.677 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Formula [FAamino_oxo(6:0/2:0)]2-amino-3-oxo-hexanedioicacid 3.621 174.04022 C6 H9 N O5 2,5-dichlorobenzoquinone 3.565 174.93674 C6 H2 Cl2 O2 2_4-Dinitrophenylhydrazine 0.906 197.03116 C6 H6 N4 O4 2-Amino-4-hydroxypteridine-6-carbaldehyde 1.304 190.03654 C7 H5 N5 O2 2-oxo-1_2-dihydroquinoline-4-carboxylate_R1 0.921 188.03479 C10 H7 N O3 2-oxo-1_2-dihydroquinoline-4-carboxylate_R2 1.381 188.03477 C10 H7 N O3 3-hydroxy-1H-quinolin-4-one_R1 0.922 160.03963 C9 H7 N O2 3-hydroxy-1H-quinolin-4-one_R2 1.754 160.03947 C9 H7 N O2 3-Sulfamoylalanine 1.184 167.01247 C3 H8 N2 O4 S 4-Formyl-2-methoxyphenyl hydrogen sulfate 0.919 230.99686 C8 H8 O6 S 4-Hydroxy-8-(beta-D-talopyranosyloxy)-2-quinolinecarboxylic acid 1.896 366.08374 C16 H17 N O9 4-Methylene-L-glutamine 3.899 157.06114 C6 H10 N2 O3 5-Hydroxyisourate 0.922 183.01541 C5 H4 N4 O4 5-L-Glutamyl-taurine 1.202 253.05017 C7 H14 N2 O6 S 6-Acetyl-2-amino-1,7,8,9-tetrahydro-4H-pyrimido[4,5-b][1,4]diazepin-4-one 1.646 220.08382 C9 H11 N5 O2 6-Aminopenicillanate_R1 0.926 215.04942 C8 H12 N2 O3 S 6-Aminopenicillanate_R2 1.747 215.04929 C8 H12 N2 O3 S Acamprosate 2.756 180.03304 C5 H11 N O4 S Asulam 0.917 229.0287 C8 H10 N2 O4 S beta-D-glucuronamide 1.501 192.05215 C6 H11 N O6 capuride 3.298 185.12909 C9 H18 N2 O2 Cholinesulfate_R1 1.926 182.0486 C5 H13 N O4 S Cholinesulfate_R2 2.467 182.04857 C5 H13 N O4 S diethylmaleate_R1 2.534 171.06563 C8 H12 O4 diethylmaleate_R2 2.626 171.06567 C8 H12 O4 DL-Homolanthionine sulfone 1.012 267.06604 C8 H16 N2 O6 S Ethyl N-acetyl-N-butyl-beta-alaninate 1.832 214.14465 C11 H21 N O3 Hydroxyanthraquinone 0.921 223.03937 C14 H8 O3 L-Homocysteicacid 0.917 182.0125 C4 H9 N O5 S L-Threonicacid 1.726 135.0302 C4 H8 O5 nitecapone 0.912 264.05182 C12 H11 N O6 N-Undecanoylglycine 1.505 242.17615 C13 H25 N O3 Pantothenicacid(VitaminB5) 3.772 218.10333 C9 H17 N O5 Sulfanilic acid 1.863 172.0067 C6 H7 N O3 S Sulfate 3.483 194.92683 H2 O4 S Sulfosalicylic Acid 0.916 216.98108 C7 H6 O6 S Tauropine 0.911 196.02814 C5 H11 N O5 S tyramine sulfate_R1 1.153 216.0337 C8 H11 N O4 S tyramine sulfate_R2 1.217 216.03317 C8 H11 N O4 S Xanthopterin-B2 1.118 237.06296 C9 H10 N4 O4 Xanthurenicacid_R1 0.927 204.02986 C10 H7 N O4 Xanthurenicacid_R2 1.021 204.02986 C10 H7 N O4 Xanthurenicacid_R3 1.755 204.02974 C10 H7 N O4 METABOLITES_END #END