#METABOLOMICS WORKBENCH mlafrano_20140711_9912781_mwtab.txt DATATRACK_ID:141 STUDY_ID:ST000095 ANALYSIS_ID:AN000151 PROJECT_ID:PR000087 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Dysfunctional lipid metabolism underlies the effect of perinatal DDT exposure PR:PROJECT_TITLE the development of metabolic syndrome PR:PROJECT_TYPE Animal chemical exposure and feeding study PR:PROJECT_SUMMARY This study evaluated the effect of perinatal DDT exposure on metabolic syndrome PR:PROJECT_SUMMARY mice exposed to doses that mimicked human exposure. The samples tested were PR:PROJECT_SUMMARY 2 groups: 1) a group with perinatal exposure to DDT consuming a high fat diet PR:PROJECT_SUMMARY 2) a control group consuming a high fat diet (CHF). Results indicated the DDHF PR:PROJECT_SUMMARY had comparatively elevated plasma concentrations of glucose, insulin, PR:PROJECT_SUMMARY and triglycerides, CYP7A1 gene expression, homeostasis model assessment- PR:PROJECT_SUMMARY resistance (HOMA-IR), and fat mass (%), as well as reduced thermogenesis. These PR:PROJECT_SUMMARY suggest perinatal DDT exposure may cause the co-occurrence of conditions PR:PROJECT_SUMMARY with metabolic syndrome, including glucose intolerance, hyperinsulinemia, and PR:PROJECT_SUMMARY impaired thermogenesis, and obesity. PR:INSTITUTE University of California, Davis PR:DEPARTMENT Environmental Toxicology PR:LABORATORY La Merrill PR:LAST_NAME La Merrill PR:FIRST_NAME Michele PR:ADDRESS 1 Shields Ave, Davis CA PR:EMAIL mlamerrill@ucdavis.edu PR:PHONE +1-530-754-7254 PR:FUNDING_SOURCE National Institute of Health (R00 ES019919, R03 DK082724, U24 DK092993, U24 PR:FUNDING_SOURCE T32 ES007059, and P60 DK020541), the American Diabetes Association, and PR:FUNDING_SOURCE intramural Project 5306-51530-019-00D #STUDY ST:STUDY_TITLE Dysfunctional lipid metabolism underlies the effect of perinatal DDT exposure ST:STUDY_TITLE the development of metabolic syndrome ST:STUDY_TYPE Chemical dosage and feeding study ST:STUDY_SUMMARY Targeted metabolomic analysis of bile acids was performed on 15 mouse liver ST:STUDY_SUMMARY collected from mice euthanized at 9 months following consumption of a high fat ST:STUDY_SUMMARY w/o perinatal DDT exposure. Funded by the National Institute of Health (R00 ST:STUDY_SUMMARY R03 DK082724, U24 DK092993, U24 DK097154, T32 ES007059, and P60 DK020541), the ST:STUDY_SUMMARY Diabetes Association, and USDA-ARS intramural Project 5306-51530-019-00D. ST:STUDY_SUMMARY were analyzed by UPLC-MS/MS using a Waters Acquity UPLC and detected on an API ST:STUDY_SUMMARY QTrap (AB Sciex, Framingham, MA, USA) by multiple reaction monitoring (MRM) ST:STUDY_SUMMARY negative mode electrospray ionization. ST:INSTITUTE University of California, Davis ST:DEPARTMENT U.S.D.A. Western Human Nutrition Research Center ST:LABORATORY Newman ST:LAST_NAME John ST:FIRST_NAME Newman ST:ADDRESS 430 W. Health Sciences Dr., Davis, CA 95616 ST:EMAIL john.newman@ars.usda.gov ST:PHONE +1-530-752-1009 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 15 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:AGE_OR_AGE_RANGE 9 months SU:GENDER Female SU:ANIMAL_HOUSING Singly housed SU:ANIMAL_FEED High fat diet (HFD, 4.73 kcal/g, 20% protein, 35% carbohydrate, and 45% fat per SU:ANIMAL_FEED D12451, Research Diets, New Brunswick, NJ) SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - T12-4 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T18-3 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T26-6 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T30-2 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T31-6 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T36-4 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T43-1 Treatment:DDT SUBJECT_SAMPLE_FACTORS - T52-6 Treatment:DDT SUBJECT_SAMPLE_FACTORS - V32-6 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V38-9 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V44-2 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V47-6 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V69-3 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V77-5 Treatment:OIL SUBJECT_SAMPLE_FACTORS - V78-6 Treatment:OIL #COLLECTION CO:COLLECTION_SUMMARY - CO:COLLECTION_PROTOCOL_FILENAME Newman_Lab_Bile_Acids_Extraction_&_Analysis_Protocol_La_Merrill_WCMC_P&F.pdf CO:SAMPLE_TYPE Liver tissue CO:COLLECTION_METHOD Dissection CO:STORAGE_CONDITIONS - 80°C CO:STORAGE_VIALS Eppendorf tubes #TREATMENT TR:TREATMENT_SUMMARY 1.7 mg DDT/kg body weight was administered to dams daily on gestational day 11 TR:TREATMENT_SUMMARY postnatal day 5 and litters were culled to 6 mice at postnatal day 6. Mice were TR:TREATMENT_SUMMARY screened from 2-6 months when they consumed a high fat diet. Mice were TR:TREATMENT_SUMMARY at 9 months of age. TR:TREATMENT_PROTOCOL_FILENAME Newman_Lab_Bile_Acid_Protocols_La_Merrill_WCMC_P&F.pdf TR:TREATMENT Intervention TR:TREATMENT_COMPOUND p,p`-DDT, o,p`-DDT TR:TREATMENT_ROUTE Oral TR:TREATMENT_DOSE 77.2% p,p`-DDT (98.5% purity neat, AccuStandard, New Haven, CT) and 22.8% TR:TREATMENT_DOSE (100% purity neat, AccuStandard) were dissolved at a final concentration of TR:TREATMENT_DOSE g DDT mixture/L organic olive oil TR:TREATMENT_DOSEDURATION From 11.5 days post coitus to postnatal day 5 TR:TREATMENT_VEHICLE Olive oil TR:ANIMAL_ENDP_EUTHANASIA 9 months TR:ANIMAL_ENDP_TISSUE_COLL_LIST Liver #SAMPLEPREP SP:SAMPLEPREP_SUMMARY A 15 mg liver sample (n=1 female/litter in 7 VEH and 8 DDT litters) was SP:SAMPLEPREP_SUMMARY on dry ice, enriched with deuterated bile acid surrogates, butylated SP:SAMPLEPREP_SUMMARY and ethylinediaminetetraacetic acid, and extracted twice with 500 uL methanol. SP:SAMPLEPREP_SUMMARY combined extract was dried, reconstituted in 100uL 50:50 methanol:acetonitrile SP:SAMPLEPREP_SUMMARY internal standards 1-phenyl-3-hexanoic acid urea and 1-cyclohexyl-3-dodecanoic SP:SAMPLEPREP_SUMMARY urea (Sigma-Aldrich, St. Louis, MO) and filtered at 0.1 µm. SP:SAMPLEPREP_PROTOCOL_FILENAME Newman_Lab_Bile_Acid_Protocols_La_Merrill_WCMC_P&F.pdf SP:PROCESSING_METHOD Pulverization on dry ice SP:EXTRACTION_METHOD Methanol SP:EXTRACT_STORAGE Placed in 10 °C autosampler and immediately analyzed SP:SAMPLE_SPIKING Spiked with deuterated bile acid surrogates. SP:ORGAN Liver #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Targeted UPLC-MS/MS method CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity UPLC CH:INSTRUMENT_NAME detector CH:COLUMN_NAME Waters Acquity BEH C18 (100 x 2mm, 1.7um) CH:METHODS_FILENAME Newman_Lab_Bile_Acids_Extraction_&_Analysis_Protocol_La_Merrill_WCMC_P&F.pdf CH:COLUMN_TEMPERATURE 60 C CH:FLOW_GRADIENT Yes CH:FLOW_RATE 0.4mL/min CH:INTERNAL_STANDARD See protocol/methods file CH:SAMPLE_INJECTION 5 L CH:SOLVENT_A 0.1% formic acid CH:SOLVENT_B 0.1% formic acid in acetonitrile CH:ANALYTICAL_TIME 16 min CH:WEAK_WASH_SOLVENT_NAME 20% methanol, 10% isopropanol CH:WEAK_WASH_VOLUME 600 L CH:STRONG_WASH_SOLVENT_NAME 50:50 Acetonitrile:Methanol CH:STRONG_WASH_VOLUME 600 L CH:SAMPLE_LOOP_SIZE 17 L #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Newman AN:ACQUISITION_DATE 41703 AN:SOFTWARE_VERSION AB Sciex Analyst version 1.6.2 AN:OPERATOR_NAME Michael La Frano AN:DETECTOR_TYPE API 4000 QTrap (AB Sciex, Framingham, MA, USA) AN:DATA_FORMAT .mzML #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME ABI Sciex API 6500 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:ION_SOURCE_TEMPERATURE See protocol/methods file MS:ION_SPRAY_VOLTAGE See protocol/methods file #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS % abundance MS_METABOLITE_DATA_START Samples T12-4 T12-4 T18-3 T18-3 T26-6 T26-6 T30-2 T30-2 T31-6 T31-6 T36-4 T36-4 T43-1 T43-1 T52-6 T52-6 V32-6 V32-6 V38-9 V38-9 V44-2 V44-2 V47-6 V47-6 V69-3 V69-3 V77-5 V77-5 V78-6 V78-6 Factors Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:DDT Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Treatment:OIL Chenodeoxycholic acid 0.1070 0.0584 0.0561 0.0076 0.1920 0.2040 0.1520 0.0532 0.1060 0.0293 0.0338 0.0494 0.2120 0.6220 Cholic acid 413.0000 236.0000 56.3000 104.0000 20.0000 89.8000 159.0000 99.5000 93.2000 78.0000 134.0000 41.9000 86.4000 500.0000 500.0000 Deoxycholic acid 0.3300 0.3500 0.2940 0.2960 0.0640 0.7480 0.6580 0.2020 0.2530 0.5030 0.1890 0.0612 0.1510 1.5300 2.1500 Dihydroxycholestanoic acid 0.9571 0.8338 0.1850 0.5174 0.0000 0.6836 1.0000 0.6139 0.1619 0.5282 0.0311 0.0579 0.3244 0.8606 0.5764 Glychochenodeoxycholic acid 0.0557 0.0120 0.0060 0.0067 0.0062 0.0238 0.0184 0.0180 0.0052 0.0134 0.0119 0.0092 0.0050 0.0045 0.0158 Glycocholic acid 3.2600 0.4530 0.4140 0.3390 0.3700 0.4100 0.3580 0.3650 0.2050 0.1750 0.7550 0.3710 0.3370 0.2190 0.7740 Glycodeoxycholic acid 0.0223 0.0379 0.0176 0.0188 0.0131 0.0318 0.0213 0.0179 0.0121 0.0280 0.0152 0.0120 0.0071 0.0234 0.0493 Glycohyocholic acid 1.0000 0.1753 0.0480 0.0753 0.0825 0.2086 0.1384 0.1765 0.0968 0.0055 0.2503 0.0000 0.1253 0.1146 0.2919 Glycohyodeoxycholic acid 0.0399 0.0172 0.0068 0.0053 0.0040 0.0092 0.0052 0.0100 0.0051 0.0055 0.0146 0.0066 0.0046 0.0082 0.0162 Glycolithocholic acid 0.0083 0.0211 0.0233 0.0194 0.0147 0.0148 0.0327 0.0696 0.0171 0.0175 0.0153 0.0087 0.0085 0.0135 0.0069 Glycoursodeoxycholic acid 0.0903 0.0123 0.0137 0.0148 0.0113 0.0195 0.0164 0.0142 0.0069 0.0129 0.0191 0.0192 0.0078 0.0081 0.0268 Hyocholic acid 0.6924 0.3883 0.4282 0.0000 0.3761 0.3292 0.3257 0.1262 0.6576 0.5238 0.3726 0.3917 0.4074 0.9114 1.0000 Hyodeoxycholic acid 0.1433 0.2061 0.0650 0.0884 0.0000 0.2681 0.2313 0.0616 0.1994 0.1555 0.2041 0.0331 0.0830 0.8827 1.0000 Murocholic acid screen 0.8457 0.2960 0.4334 0.0000 0.2833 0.2622 0.3552 0.0247 0.5433 0.1734 0.1416 0.0848 0.2748 0.9070 1.0000 Taurochenodeoxycholic acid 58.2000 5.3400 10.4000 6.1500 6.0900 23.8000 16.8000 16.4000 5.3200 12.5000 6.1000 8.1200 6.5100 7.6100 12.5000 Taurocholic acid 848.0000 210.0000 188.0000 203.0000 115.0000 204.0000 176.0000 208.0000 89.8000 121.0000 283.0000 176.0000 183.0000 219.0000 428.0000 Taurodeoxycholic acid 32.4000 18.6000 24.9000 15.2000 10.0000 39.2000 24.3000 17.6000 10.5000 29.8000 8.2300 10.0000 6.0300 27.3000 45.2000 Taurohyocholic acid 1.0000 0.1960 0.0585 0.1587 0.0174 0.2763 0.1637 0.1301 0.0635 0.0367 0.0865 0.0000 0.1581 0.2657 0.2390 Taurolithocholic acid 3.8500 3.0800 1.5500 1.2800 1.1700 2.8500 3.5700 1.0100 0.8240 3.4800 0.8320 1.0300 0.9880 1.2800 1.8800 Tauroursodeoxycholic acid 66.3000 15.1000 12.5000 14.2000 8.3500 22.2000 16.5000 14.1000 7.1000 16.8000 19.6000 13.0000 9.4500 14.0000 34.3000 Tauro-α-Muricholic acid 88.7000 21.6000 10.7000 15.8000 13.0000 31.2000 22.3000 17.9000 7.9100 18.0000 20.9000 17.5000 15.1000 15.4000 27.3000 Tauro-β-Muricholic acid 268.0000 79.9000 40.7000 62.2000 46.1000 65.5000 44.4000 46.4000 38.5000 45.8000 98.1000 79.2000 41.6000 76.1000 122.0000 Tauro-ω-Muricholic acid 211.0000 76.1000 34.3000 45.2000 32.0000 71.7000 39.0000 63.0000 39.1000 30.7000 66.5000 32.6000 42.5000 96.5000 82.6000 Trihydroxycholestanoic acid (screening*) 1.0000 0.0919 0.0738 0.0000 0.0444 0.0964 0.0659 0.0783 0.0223 0.0442 0.0904 0.0660 0.0934 0.0559 0.1505 Ursodeoxycholic acid 0.1030 0.1940 0.0902 0.2280 0.0184 0.3630 0.3490 0.0586 0.0941 0.3380 0.1160 0.0767 0.0957 0.8390 1.1700 α-Muricholic acid 1.9500 1.5900 0.3040 0.5040 0.1950 1.5800 1.3400 0.3150 0.8150 1.0600 0.8800 0.4790 0.5890 2.6100 3.3100 β-Muricholic acid 2.3900 3.1400 1.0000 1.6200 0.3830 3.2400 2.8500 0.8260 2.5300 2.3600 2.3400 1.5000 1.0800 7.8900 8.3500 ω-Muricholic acid 2.2300 1.9900 0.5990 0.6820 0.1670 1.9300 1.3100 0.5440 1.6600 0.8180 1.2400 0.3240 0.7330 6.4000 4.6300 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type Chenodeoxycholic acid 44135443 HMDB00518 UCDavis_Newman_ID Cholic acid 221493 HMDB00619 UCDavis_Newman_ID Deoxycholic acid 222528 HMDB00626 UCDavis_Newman_ID Dihydroxycholestanoic acid HMDB01974 UCDavis_Newman_ID Glychochenodeoxycholic acid HMDB00637 UCDavis_Newman_ID Glycocholic acid 755 HMDB00138 UCDavis_Newman_ID Glycodeoxycholic acid 3035026 HMDB00631 UCDavis_Newman_ID Glycohyocholic acid Glycohyodeoxycholic acid 114611 Glycolithocholic acid 115245 HMDB00698 UCDavis_Newman_ID Glycoursodeoxycholic acid HMDB00708 UCDavis_Newman_ID Hyocholic acid 92805 HMDB00865 UCDavis_Newman_ID Hyodeoxycholic acid 9963687 Murocholic acid screen HMDB00811 UCDavis_Newman_ID Taurochenodeoxycholic acid 387316 HMDB00951 UCDavis_Newman_ID Taurocholic acid 444349 HMDB00036 UCDavis_Newman_ID Taurodeoxycholic acid 10594 HMDB00896 UCDavis_Newman_ID Taurohyocholic acid Taurolithocholic acid 53477716 HMDB00722 UCDavis_Newman_ID Tauroursodeoxycholic acid 9848818 HMDB00874 UCDavis_Newman_ID Tauro-α-Muricholic acid Tauro-β-Muricholic acid HMDB00932 UCDavis_Newman_ID Tauro-ω-Muricholic acid 21124703 Trihydroxycholestanoic acid (screening*) HMDB02163 UCDavis_Newman_ID Ursodeoxycholic acid 31401 HMDB00946 UCDavis_Newman_ID α-Muricholic acid 5283852 HMDB00506 UCDavis_Newman_ID β-Muricholic acid 5283853 HMDB00415 UCDavis_Newman_ID ω-Muricholic acid 5283851 HMDB00364 UCDavis_Newman_ID METABOLITES_END #END