#METABOLOMICS WORKBENCH akane_kanasaki_20210119_224723 DATATRACK_ID:2405 STUDY_ID:ST001668 ANALYSIS_ID:AN002722 PROJECT_ID:PR001071 VERSION 1 CREATED_ON January 28, 2021, 1:16 pm #PROJECT PR:PROJECT_TITLE D-Allulose effects on metabolomics profile in rodents PR:PROJECT_SUMMARY To determine the metabolomics profile after D-allulose intake. PR:INSTITUTE Matsutani Chemical Industry Co., Ltd. PR:LAST_NAME Kanasaki PR:FIRST_NAME Akane PR:ADDRESS 5-3 Kita-Itami, Itami, Hyogo, 664-8508, Japan PR:EMAIL akane-kanasaki@matsutani.co.jp PR:PHONE 81-72-771-2052 #STUDY ST:STUDY_TITLE D-Allulose effects on hepatic metabolomics profile in rodents ST:STUDY_SUMMARY To determine the hepatic metabolomics profile after D-allulose intake. ST:INSTITUTE Matsutani Chemical Industry Co., Ltd. ST:LAST_NAME Kanasaki ST:FIRST_NAME Akane ST:ADDRESS 5-3 Kita-Itami, Itami, Hyogo, 664-8508, Japan ST:EMAIL akane-kanasaki@matsutani.co.jp ST:PHONE 81-72-771-2052 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 #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 - C1 Diet:control SUBJECT_SAMPLE_FACTORS - C2 Diet:control SUBJECT_SAMPLE_FACTORS - C3 Diet:control SUBJECT_SAMPLE_FACTORS - C4 Diet:control SUBJECT_SAMPLE_FACTORS - C5 Diet:control SUBJECT_SAMPLE_FACTORS - C6 Diet:control SUBJECT_SAMPLE_FACTORS - P1 Diet:D-Allulose SUBJECT_SAMPLE_FACTORS - P2 Diet:D-Allulose SUBJECT_SAMPLE_FACTORS - P3 Diet:D-Allulose SUBJECT_SAMPLE_FACTORS - P4 Diet:D-Allulose SUBJECT_SAMPLE_FACTORS - P5 Diet:D-Allulose SUBJECT_SAMPLE_FACTORS - P6 Diet:D-Allulose #COLLECTION CO:COLLECTION_SUMMARY The livers were collected and then flash-frozen in liquid N2 CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY Seven-week old male Wistar rats were fed AIN-93G-based diets with/without 3% TR:TREATMENT_SUMMARY D-allulose, ad libitum for 4 weeks. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For CE-TOFMS, The frozen tissue was plunged into 50% acetonitrile/Milli-Q water SP:SAMPLEPREP_SUMMARY containing internal standards. The tissues were homogenized thrice, and then SP:SAMPLEPREP_SUMMARY centrifuged. The upper aqueous layer was centrifugally filtered to remove the SP:SAMPLEPREP_SUMMARY proteins. The filtrate was then centrifugally concentrated and re-suspended in SP:SAMPLEPREP_SUMMARY 50 µL of Milli-Q water for CE-MS analysis. For LC-TOFMS, the frozen tissue was SP:SAMPLEPREP_SUMMARY plunged into 1% formic acid/acetonitrile containing the internal standard SP:SAMPLEPREP_SUMMARY solution. Tissue was homogenized thrice, and the mixture was yet again SP:SAMPLEPREP_SUMMARY homogenized after adding Milli-Q water and then centrifuged. The supernatant was SP:SAMPLEPREP_SUMMARY collected and 500 1% formic acid/acetonitrile and Milli-Q water were added to SP:SAMPLEPREP_SUMMARY the precipitate. Homogenization and centrifugation were performed as described SP:SAMPLEPREP_SUMMARY previously, and the supernatant was mixed with previously collected one. The SP:SAMPLEPREP_SUMMARY mixed supernatant was filtrated to remove the proteins and phospholipids. The SP:SAMPLEPREP_SUMMARY filtrate was then desiccated and dissolved in 200 µL of isopropanol/Milli-Q SP:SAMPLEPREP_SUMMARY water for LC-MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1200SL CH:COLUMN_NAME ODS column (2×50 mm, 2 μm) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6230 TOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS - #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Relative Area MS_METABOLITE_DATA_START Samples C1 C2 C3 C4 C5 C6 P1 P2 P3 P4 P5 P6 Factors Diet:control Diet:control Diet:control Diet:control Diet:control Diet:control Diet:D-Allulose Diet:D-Allulose Diet:D-Allulose Diet:D-Allulose Diet:D-Allulose Diet:D-Allulose Estradiol-17alpha 0.000281 0.000188 0.000316 0.000177 0.000186 0.00024 0.000203 0.000238 0.000216 Androstenedione 0.000225 0.000143 0.000174 0.000144 0.000135 0.000125 0.000205 0.000144 0.000109 0.000099 0.000108 Testosterone 0.00019 0.000192 0.000226 0.00046 0.000317 0.000173 0.000324 0.000927 0.00032 0.000301 0.000215 Palmitoyl-EA 0.0009 0.001182 0.001135 0.001559 0.001103 0.001029 0.001097 0.001581 0.001057 0.001232 0.001057 0.001226 Sphingosine 0.0142 0.0115 0.0178 0.012 0.0136 0.0157 0.0122 0.0149 0.0129 0.0103 0.0139 0.0137 Sphinganine 0.00237 0.00226 0.00235 0.00203 0.00261 0.00258 0.0022 0.00263 0.0029 0.00252 0.00237 0.00268 306.2_8.1 0.000085 0.000123 0.000093 0.000112 0.000058 0.000077 0.000092 306.2_8.3 0.000144 0.000164 0.000166 0.000127 0.000151 0.000127 0.000145 0.000136 0.000131 0.000148 0.000114 0.000175 306.2_8.7 0.00013 0.000092 0.000085 0.000068 CAR 10:0 0.000316 0.00058 0.000491 0.000405 0.00069 0.000325 0.00079 0.000652 0.000751 0.000747 0.000749 0.000887 317.2_10.6 0.000372 0.000432 0.000348 0.000391 0.000315 0.000439 0.000354 317.3_11.1 0.00033 0.000267 0.000494 5alpha-Pregnane-3,20-dione 0.000656 0.000514 0.000551 0.000718 0.000661 0.000571 0.00051 0.000677 318.3_10.9 0.00132 0.00158 0.00158 0.00138 0.00113 0.00164 0.00139 0.00159 0.0015 0.00103 0.00141 0.00127 318.3_10.7 0.000567 0.000646 0.000566 0.00068 Stearoyl-EA 0.000315 0.000671 0.000747 0.001029 0.000416 0.000222 0.000543 0.000297 0.000381 0.000225 17alpha-Hydroxyprogesterone 0.000061 0.000071 0.000095 0.000098 0.000036 11-Deoxycorticosterone 0.000096 0.000079 0.00007 0.000122 0.000138 0.000079 Testosterone acetate 0.000098 0.000158 0.000109 0.000093 0.000226 0.000102 0.000149 0.000152 PGB2 0.000037 0.000068 0.000085 0.000058 CAR 12:1 0.000511 0.000529 0.00063 0.000495 0.000739 0.000348 0.000895 0.000769 0.000981 0.00103 0.000967 0.001017 CAR 12:0 0.000474 0.00075 0.000832 0.000577 0.000823 0.000573 0.000969 0.000774 0.001131 0.001133 0.000989 0.001051 11-Deoxycortisol Corticosterone 21-Deoxycortisol 0.000088 0.000065 0.000116 0.000104 0.000101 0.000072 0.000152 0.000089 0.000107 0.000102 0.000214 356.3_9.7 0.000065 0.00007 0.000152 356.3_9.9 0.000077 0.000095 0.00016 0.000072 0.000119 0.000182 CAR 14:3 0.000132 0.000062 0.000095 0.000146 0.000139 0.000055 0.000111 0.000139 0.00015 0.00013 0.000094 0.000119 367.3_15.9 0.00023 0.000203 0.00029 0.000221 0.000211 0.000365 367.3_15.5 0.000355 0.00049 0.000398 0.000372 0.000463 0.000479 0.000577 0.000667 0.000531 0.000588 0.000584 0.000715 367.3_16.1 0.000435 0.00032 0.00025 0.000383 0.000496 0.000355 0.000292 0.000455 0.000436 0.00053 0.00043 0.000461 368.3_9.7 0.000067 0.000072 0.00004 368.3_9.8 0.000409 0.000366 0.000387 0.000343 0.000625 0.000297 0.000588 0.000674 0.001061 0.000891 0.000694 0.000846 370.3_10.2 0.00041 0.000509 0.000728 0.00034 0.000435 0.000431 0.000691 0.000666 0.001006 0.000828 0.000579 0.000679 370.3_10.3 0.000607 0.000529 0.000642 0.000509 0.000781 0.000481 0.000979 0.000802 0.001202 0.001329 0.00097 0.001105 Cholesterol 0.0356 0.0396 0.0406 0.0395 0.0401 0.0379 0.039 0.0468 0.044 0.0423 0.0434 0.0448 PGE1 PGD1 0.00013 0.000087 0.000168 0.000241 CAR 14:0 0.0017 0.00182 0.00233 0.00152 0.00216 0.00155 0.00232 0.0023 0.00327 0.00309 0.00259 0.0029 FMN 0.000283 0.000162 0.000239 0.000135 0.000183 0.000129 0.000204 0.000158 0.000232 0.000145 0.00019 MG 20:4 0.00362 0.00342 0.00304 0.00315 0.00221 0.0034 0.00259 0.00388 0.00276 0.00238 0.00215 0.00287 383.4_16.4 0.000103 0.000148 0.000094 0.000062 0.000175 0.000132 0.000061 0.000176 0.000116 0.000055 383.4_16.2 0.00276 0.00283 0.00384 0.00325 0.00318 0.00299 0.00355 0.00372 0.00444 0.00369 0.00356 0.00423 386.3_11.0 0.000244 0.000204 0.000188 0.000168 0.000127 0.000191 0.000261 0.00022 0.000216 0.000255 0.000234 0.000174 386.3_11.1 0.000173 0.000192 0.00017 0.00011 0.000218 0.000166 0.000174 0.000244 0.000254 0.000261 0.000178 0.000214 5alpha-Cholestanone 0.000043 0.00007 0.000031 0.000094 395.4_16.3 0.000151 0.000127 0.000186 0.000157 0.000135 0.00018 0.00013 0.000175 0.000206 0.000142 0.00018 0.000167 395.4_16.1 0.001032 0.000961 0.001402 0.001025 0.000978 0.001055 0.001208 0.001397 0.001316 0.001266 0.001439 0.001388 CAR 16:2 0.000671 0.000684 0.00063 0.000625 0.00088 0.000565 0.001038 0.001048 0.001867 0.00121 0.001139 0.001173 Sitosterol 0.00257 0.00267 0.00378 0.00269 0.00262 0.00267 0.00306 0.00351 0.00386 0.00327 0.00343 0.00369 CAR 16:1 0.0031 0.00253 0.00535 0.00237 0.00306 0.00362 0.00429 0.00429 0.00645 0.00503 0.00424 0.00413 CAR 16:0 0.0129 0.0112 0.0155 0.0093 0.014 0.0125 0.0104 0.0137 0.0182 0.0187 0.012 0.0157 Cycloartenol Lanosterol 0.000105 0.000103 0.000137 0.000225 0.00012 0.000145 0.000124 CAR 17:1 0.00015 0.000135 0.000163 0.00017 0.000104 0.000126 0.000134 0.000243 0.000161 0.000146 0.000176 0.000177 414.4_11.9 0.000192 0.000182 0.000247 0.000176 0.000269 0.000202 0.000182 0.000231 0.000227 0.000276 0.000149 0.000294 414.4_11.7 0.000251 0.000153 0.00026 0.00017 0.000245 0.000187 0.00018 0.000247 0.000334 0.000249 0.000225 0.000267 417.4_16.1 0.000963 0.000984 0.001111 0.00114 0.00117 0.001173 0.00125 0.001496 0.001424 0.001146 0.001101 0.000912 417.4_15.7 0.000131 0.000076 0.000112 CAR 18:2 0.00567 0.00329 0.00524 0.00362 0.00613 0.00523 0.00644 0.0071 0.01076 0.00773 0.00631 0.00882 CAR 18:1 0.012 0.0086 0.0155 0.0072 0.0117 0.0118 0.0131 0.0141 0.02 0.0163 0.0119 0.0168 428.4_12.1 0.000111 0.000184 0.000103 0.000159 0.000184 0.000171 0.000082 428.4_12.2 0.00741 0.00689 0.00783 0.00532 0.00633 0.00669 0.00457 0.00612 0.00798 0.00758 0.00613 0.00771 alpha-Tocopherol 0.0291 0.0283 0.0355 0.0352 0.0338 0.0359 0.0278 0.0354 0.0351 0.0295 0.0333 0.033 LPC 12:0 0.000048 0.000046 0.000088 0.000171 0.000115 0.000061 0.000133 CAR 20:1 0.0032 0.00174 0.00456 0.0012 0.00192 0.00199 0.00246 0.00254 0.00274 0.00268 0.00198 0.00309 CAR 20:0 0.000579 0.000452 0.00065 0.000369 0.000364 0.00043 0.000451 0.000416 0.00032 0.000475 CAR 22:0 0.000193 0.000113 0.000172 0.000167 0.000166 0.000185 0.000178 0.000187 0.000163 0.000172 0.000206 0.00025 490.4_16.5 0.000102 0.000142 0.000144 0.000145 0.000078 0.00012 0.000105 0.000119 0.000107 0.000118 0.00006 SM 18:1;O2/16:0 0.000622 0.000669 0.00068 0.000576 0.000581 0.000676 0.000741 0.000861 0.000671 0.000659 0.000695 0.000718 731.6_16.6 0.000189 0.000232 0.000273 0.000172 0.000246 0.000216 0.00025 0.000359 0.000227 0.000174 0.000256 731.6_16.2 0.000233 0.000134 0.000155 790.6_16.9 0.000134 0.000156 0.000162 0.000109 0.000125 0.000085 0.000188 0.00012 0.0001 790.7_17.1 0.00023 0.000213 0.00033 0.000249 0.000346 0.000343 0.000383 0.000283 0.000351 0.000283 0.000364 0.00044 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PubChem CID m/z Estradiol-17alpha 68570 273.192 Androstenedione 6128 287.206 Testosterone 6013 289.211 Palmitoyl-EA 4671 300.293 Sphingosine 5280335 300.294 Sphinganine 91486 302.309 306.2_8.1 5756 306.2 306.2_8.3 5756 306.2 306.2_8.7 68929 306.2 CAR 10:0 316.3 317.2_10.6 8956 317.2 317.3_11.1 8955 317.3 5alpha-Pregnane-3,20-dione 317.3 318.3_10.9 122121 318.3 318.3_10.7 122121 318.3 Stearoyl-EA 27902 328.3 17alpha-Hydroxyprogesterone 62386166 331.2 11-Deoxycorticosterone 440698 331.2 Testosterone acetate 92145 331.2 PGB2 5280881 335.2 CAR 12:1 342.3 CAR 12:0 344.3 11-Deoxycortisol Corticosterone 21-Deoxycortisol 347.2 356.3_9.7 356.3 356.3_9.9 356.3 CAR 14:3 366.3 367.3_15.9 367.3 367.3_15.5 439577 367.3 367.3_16.1 439423 367.3 368.3_9.7 368.3 368.3_9.8 368.3 370.3_10.2 370.3 370.3_10.3 370.3 Cholesterol 5997 370.4 PGE1 PGD1 372.3 CAR 14:0 372.3 FMN 493570 377.1 MG 20:4 5282280 379.3 383.4_16.4 173183 383.4 383.4_16.2 173183 383.4 386.3_11.0 386.3 386.3_11.1 386.3 5alpha-Cholestanone 92128 387.4 395.4_16.3 5280794 395.4 395.4_16.1 5280794 395.4 CAR 16:2 396.3 Sitosterol 222284 397.4 CAR 16:1 398.3 CAR 16:0 11953816 400.3 Cycloartenol Lanosterol 409.4 CAR 17:1 412.3 414.4_11.9 414.4 414.4_11.7 414.4 417.4_16.1 92729 417.4 417.4_15.7 92729 417.4 CAR 18:2 424.3 CAR 18:1 426.4 428.4_12.1 428.4 428.4_12.2 428.4 alpha-Tocopherol 14985 431.4 LPC 12:0 440.3 CAR 20:1 454.4 CAR 20:0 456.4 CAR 22:0 484.4 490.4_16.5 86472 490.4 SM 18:1;O2/16:0 5283590 703.6 731.6_16.6 5283588 731.6 731.6_16.2 5283588 731.6 790.6_16.9 94190 790.6 790.7_17.1 94190 790.7 METABOLITES_END #END