#METABOLOMICS WORKBENCH n_hiraoka_20240509_004912 DATATRACK_ID:4828 STUDY_ID:ST003298 ANALYSIS_ID:AN005404 PROJECT_ID:PR002049 VERSION 1 CREATED_ON May 14, 2024, 1:44 am #PROJECT PR:PROJECT_TITLE Annual changes on metabolomics profile in latex PR:PROJECT_SUMMARY To clarify seasonal changes in metabolites contained in latex, we determine the PR:PROJECT_SUMMARY metabolomics profile in latex of Hevea brasiliensis throughout the year. As a PR:PROJECT_SUMMARY result, a total of more than 400 metabolites had difference not only in amount, PR:PROJECT_SUMMARY but also in existence. PR:INSTITUTE Sumitomo Riko Co., Ltd. PR:DEPARTMENT Fundamental Material Development Laboratory PR:LAST_NAME HIRAOKA PR:FIRST_NAME Nobuyuki PR:ADDRESS 1, Higashi 3-chome, Komaki-shi, Aichi, 485-8550, Japan PR:EMAIL nobuyuki.hiraoka@jp.sumitomoriko.com PR:PHONE 0568-77-2132 #STUDY ST:STUDY_TITLE Annual changes on metabolomics profile in latex ST:STUDY_SUMMARY Natural rubber (NR) is an important material with excellent physical properties. ST:STUDY_SUMMARY Unlike synthetic rubber from petroleum, NR contains non-rubber components such ST:STUDY_SUMMARY as proteins, lipids, and metal ions. The non-rubber components are known to ST:STUDY_SUMMARY affect the properties of NR. In this study, latex samples of Hevea brasiliensis ST:STUDY_SUMMARY were collected for nine months and their metabolites were comprehensively ST:STUDY_SUMMARY analyzed by mass spectrometry. NR was made from the same latex samples used for ST:STUDY_SUMMARY the mass spectrometry, and their vulcanization, tensile and thermal-aging ST:STUDY_SUMMARY properties were assessed. These results suggest that the metabolite composition ST:STUDY_SUMMARY in the latex and the NR properties changed seasonally. Correlation analysis ST:STUDY_SUMMARY between the metabolites and the properties of NR indicated that different ST:STUDY_SUMMARY metabolites affected different properties. A regression model of NR properties ST:STUDY_SUMMARY using metabolites as the explanatory variables suggests that about five ST:STUDY_SUMMARY metabolites need to be considered when examining the relationship between ST:STUDY_SUMMARY properties and metabolites. This method, which combines comprehensive analysis ST:STUDY_SUMMARY and characterization of NR, contributes to studies aimed at elucidating how the ST:STUDY_SUMMARY superior properties of NR are brought about. ST:INSTITUTE Sumitomo Riko Co., Ltd. ST:DEPARTMENT Fundamental Material Development Laboratory ST:LAST_NAME Nobuyuki ST:FIRST_NAME Hiraoka ST:ADDRESS 1, Higashi 3-chome, Komaki-shi, Aichi 485-8550, Japan ST:EMAIL nobuyuki.hiraoka@jp.sumitomoriko.com ST:PHONE +81-568-77-2132 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Hevea brasiliensis #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 STRCH-HMT-003 TFL269 Sample source:latex | collected month:May-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-003 TFL272 Sample source:latex | collected month:Jun-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-003 TFL275 Sample source:latex | collected month:Jul-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL278 Sample source:latex | collected month:Aug-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL281 Sample source:latex | collected month:Sep-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL284 Sample source:latex | collected month:Oct-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL287 Sample source:latex | collected month:Nov-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL290 Sample source:latex | collected month:Dec-22 SUBJECT_SAMPLE_FACTORS STRCH-HMT-004 TFL293 Sample source:latex | collected month:Jan-23 #COLLECTION CO:COLLECTION_SUMMARY The latex collected on same day from some trees (Hevea brasiliensis) was mixed. CO:SAMPLE_TYPE Latex CO:STORAGE_CONDITIONS 4℃ #TREATMENT TR:TREATMENT_SUMMARY Ammonia was added to the latex to prevent coagulation. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For CE-TOFMS, the latex was plunged into methanol containing internal standards. SP:SAMPLEPREP_SUMMARY The latex sample was homogenized thrice, and then centrifuged. The upper aqueous SP:SAMPLEPREP_SUMMARY layer was centrifugally filtered to remove the proteins. The filtrate was then SP:SAMPLEPREP_SUMMARY centrifugally concentrated and re-suspended in Milli-Q water for CE-MS analysis. SP:SAMPLEPREP_SUMMARY For LC-TOFMS, the latex was plunged into 1% formic acid/acetonitrile containing SP:SAMPLEPREP_SUMMARY the internal standard solution. The latex sample was homogenized thrice, and the SP:SAMPLEPREP_SUMMARY mixture was yet again homogenized after adding Milli-Q water and then SP:SAMPLEPREP_SUMMARY centrifuged. The supernatant was collected and 1% formic acid/acetonitrile and SP:SAMPLEPREP_SUMMARY Milli-Q water were added to the precipitate. Homogenization and centrifugation SP:SAMPLEPREP_SUMMARY were performed as described previously, and the supernatant was mixed with SP:SAMPLEPREP_SUMMARY previously collected one. The mixed supernatant was filtrated to remove the SP:SAMPLEPREP_SUMMARY proteins and phospholipids. The filtrate was then desiccated and dissolved in 1% SP:SAMPLEPREP_SUMMARY isopropanol/Milli-Q water for LC-MS analysis. SP:PROCESSING_STORAGE_CONDITIONS Room temperature SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1200 series RRLC system SL CH:COLUMN_NAME Osaka Soda capcell pak c18 if (50 x 2mm,3um) CH:SOLVENT_A 100% Water; 0.1% Formic acid CH:SOLVENT_B 65% Isopropanol, 30% Acetonitrile 5% Water; 0.1% Formic acid, 2 mM Ammonium CH:SOLVENT_B formate CH:FLOW_GRADIENT 0-0.5 min: B 1%, 0.5-13.5 min: B 1-100%, 13.5-20 min: B 100% CH:FLOW_RATE 0.3 mL / min CH:COLUMN_TEMPERATURE 40℃ #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6210 TOF MS:INSTRUMENT_TYPE Other MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using MS:MS_COMMENTS automatic integration software MasterHands (Keio University, Tsuruoka, Japan). MS:MS_COMMENTS Signal peaks corresponding to isotopomers, adduct ions, and other product ions MS:MS_COMMENTS of known metabolites were excluded, and the remaining peaks were annotated MS:MS_COMMENTS according to HMT’s metabolite database based on their m/z values and RTs. MS:MS_COMMENTS Areas of the annotated peaks were then normalized to internal standards and MS:MS_COMMENTS sample amount in order to obtain relative levels of each metabolite. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Relative Area MS_METABOLITE_DATA_START Samples TFL269 TFL272 TFL275 TFL278 TFL281 TFL284 TFL287 TFL290 TFL293 Factors Sample source:latex | collected month:May-22 Sample source:latex | collected month:Jun-22 Sample source:latex | collected month:Jul-22 Sample source:latex | collected month:Aug-22 Sample source:latex | collected month:Sep-22 Sample source:latex | collected month:Oct-22 Sample source:latex | collected month:Nov-22 Sample source:latex | collected month:Dec-22 Sample source:latex | collected month:Jan-23 1,2-Distearoyl-glycero-3-phosphocholine 9.82E-05 4.09E-05 6.94E-05 2.85E-05 2.30E-05 6.73E-05 4.28E-05 2.34E-05 2.45E-05 15-Deoxy-Δ12,14-prostaglandin J2 3.97E-05 2.32E-05 3.60E-05 2.85E-05 2.91E-05 5.44E-05 17α-Estradiol 3.59E-05 17α-Hydroxyprogesterone 1.01E-05 2.68E-05 6.93E-06 3.56E-05 2.93E-05 9.38E-06 3.79E-05 9.62E-06 1-Deoxysphinganine 2.62E-05 2.24E-05 1-Oleoyl-glycero-3-phosphocholine 1.34E-05 7.82E-06 1.08E-05 1-Palmitoyl-glycero-3-phosphocholine 2.30E-05 2.07E-05 2.42E-05 1-Stearoyl-glycero-3-phosphocholine 1.07E-05 1.32E-05 8.16E-06 1.30E-05 1.67E-05 1.72E-05 22-Hydroxycholesterol 9.15E-06 3.37E-05 2.59E-05 2.33E-05 2.69E-05 5.27E-05 3β-Hydroxy-5-cholestenoic acid 8.58E-05 0.000154679 0.00010338 0.000231171 5α-Cholestan-3-one 2.56E-05 1.62E-05 1.65E-05 6-Gingerol 2.48E-05 2.11E-05 1.50E-05 2.45E-05 7-Dehydrocholesterol 1.05E-05 5.70E-06 6.58E-06 Abietic acid 3.98E-05 4.39E-05 Acylcarnitine(18:1) 1.67E-05 1.16E-05 2.46E-05 1.34E-05 9.64E-06 Acylcarnitine(20:1) 0.000115741 0.00014374 0.000109597 5.51E-05 9.19E-05 0.000124212 AEA(18:3) 0.00554794 0.003895564 0.008478444 0.005908653 0.005012747 0.001997262 0.003013898 0.003404332 0.007695646 AEA(20:3) 5.53E-05 0.000102681 0.000296851 0.000178908 0.000130392 5.36E-05 6.93E-05 4.14E-05 0.000113562 AEA(22:6) 2.38E-05 2.57E-05 Anandamide 2.56E-05 4.93E-05 0.000110448 8.90E-05 6.41E-05 2.88E-05 2.93E-05 4.38E-05 Bisacurone 1.56E-05 4.08E-05 9.84E-06 4.82E-05 Campesterol 0.000256747 0.000241346 0.000214196 0.000409017 0.000359658 0.000324091 0.000362397 0.000344369 0.000369769 Coenzyme Q10 7.62E-05 7.07E-05 4.55E-05 0.000288819 7.50E-05 9.37E-05 6.38E-05 0.000154356 4.70E-05 Dehydrozingerone 0.000150868 0.000223941 0.000177186 0.000215799 0.000165216 0.000148019 Deoxycorticosterone 8.63E-06 Desmosterol 3.55E-05 2.44E-05 2.19E-05 Eleutheroside B 7.75E-05 5.64E-05 6.02E-05 0.000164925 0.000177009 0.000142011 0.000303532 0.000257328 0.000514691 Etiocholan-3α-ol-17-one 9.65E-06 Flavanone 3.45E-05 3.00E-05 5.10E-05 3.51E-05 4.51E-05 4.35E-05 Glabridin 1.22E-05 1.01E-05 1.76E-05 2.64E-05 9.66E-06 2.00E-05 Glucosylceramide(d18:1/24:1) 7.27E-05 6.16E-05 4.70E-05 3.52E-05 3.61E-05 3.30E-05 Hecogenin 2.20E-05 8.27E-06 4.42E-05 5.34E-05 2.40E-05 3.05E-05 4.92E-05 4.78E-05 Hydroxyprogesterone caproate 3.55E-05 1.75E-05 1.46E-05 1.73E-05 2.04E-05 2.66E-05 2.22E-05 2.33E-05 3.08E-05 Indole-3-carboxaldehyde 0.000346562 0.000283281 0.000165937 0.000377543 0.000546885 0.000202087 0.000183866 0.000547158 0.000724694 Isofraxidin 0.001159914 0.002485421 0.001521402 0.00326369 0.001968408 0.002298823 0.000822198 0.002766726 0.001138423 Kynurenic acid 7.48E-05 3.53E-05 4.24E-05 4.49E-05 4.05E-05 7.25E-05 Linoleyl ethanolamide 0.018240409 0.009945005 0.018964921 0.011601269 0.011891713 0.005989625 0.010018628 0.01040202 0.02066717 Lutein 2.19E-05 1.35E-05 7.09E-06 N-Oleoylglycine 9.31E-05 0.000136477 6.56E-05 9.01E-05 9.55E-05 0.000120771 PAF(17:0) 1.86E-05 1.65E-05 2.24E-05 3.58E-05 1.54E-05 1.92E-05 1.83E-05 Palmitoylcarnitine 1.15E-05 1.52E-05 2.84E-05 Palmitoylcholine 0.000418003 0.000749384 0.000341226 0.000235459 0.0006011 0.000765901 Palmitoylethanolamide 0.001760459 0.000891204 0.001412902 0.000857778 0.001169279 0.000744548 0.000959646 0.001346999 0.0017256 Phytosphingosine 8.48E-05 4.61E-05 4.30E-05 2.31E-05 2.75E-05 2.33E-05 3.02E-05 2.17E-05 4.65E-05 Prostaglandin B2 1.31E-05 Retinol 1.32E-05 1.47E-05 Riboflavin 1.95E-05 2.00E-05 8.96E-06 8.27E-06 7.50E-06 Sphinganine 1.60E-05 9.08E-06 Sphinganine(d20:0) 0.000117329 8.42E-05 7.27E-05 0.000262708 0.000285269 0.000224812 0.000189923 0.000396507 0.000241231 Sphinganine(d22:0) 2.96E-05 2.41E-05 2.45E-05 2.11E-05 4.82E-05 Sphingomyelin(d18:1/16:0) 3.15E-05 3.17E-05 1.54E-05 Sphingomyelin(d18:1/18:0) 0.000115288 0.000173421 0.000385268 9.28E-05 0.000155228 Sphingosine 2.98E-05 Sphingosine(d20:1) 3.60E-05 2.49E-05 1.58E-05 4.66E-05 5.77E-05 5.89E-05 0.000126215 9.27E-05 Stearoyl ethanolamide 0.005310369 0.002594555 0.002599747 0.001942012 0.003457617 0.00200302 0.002516684 0.003758557 0.003958518 Trilaurin 4.66E-05 3.92E-05 3.53E-05 0.000104212 0.000126689 8.37E-05 0.000134628 6.66E-05 Triptolide 0.000162245 0.000229486 0.000264197 0.00040478 0.000365284 0.000335858 0.000371981 0.000423768 0.000410649 Tropic acid 5.41E-05 6.51E-05 6.44E-05 Zeaxanthin 2.76E-05 3.19E-05 2.60E-05 4.17E-05 3.53E-05 α-Tocopherol 0.001518792 0.001461756 0.000898083 0.001502106 0.001338925 0.000609369 0.001269452 0.001369599 0.001470884 α-Tocopherol acetate 3.04E-05 3.03E-05 0.000787363 0.00014289 7.85E-05 6.04E-05 6.50E-05 0.000111117 6.14E-05 β-Cryptoxanthin 0.00013686 0.000124234 6.55E-05 0.000186565 0.000140338 0.000124152 6.05E-05 0.000124269 0.000101782 β-Estradiol 3.81E-05 2.57E-05 3.42E-05 7.05E-05 8.22E-05 4.97E-05 6.39E-05 8.59E-05 2-Aminoadipic acid;O-Acetylhomoserine;Glutamic acid γ-methyl ester 0.000114513 0.000164289 0.000206355 0.000241696 0.000175382 0.000117666 0.000105761 0.00036789 0.000455573 6-Keto-prostaglandin E1 1.21E-05 1.45E-05 1.73E-05 9.51E-06 5.54E-05 9(S)-HODE 0.00067247 0.002182934 0.002667319 0.002218298 0.001283106 0.002265786 AEA(18:1);Oleoyl ethanolamide 0.004898459 0.001805175 0.002751781 0.001526435 0.002529919 0.001856031 0.00220737 0.002709095 0.003031556 AEA(22:4) 0.00016532 0.000165453 0.00042831 Ethyl arachidonate 0.000445263 0.000334646 0.000267043 0.000459618 0.000401409 0.000285361 0.000222998 0.000375507 0.000205267 Lanosterol;Cycloartenol 0.000753741 0.000708531 0.000724793 0.000980416 0.000842459 0.000666195 0.000993559 0.000894102 0.000939251 Lathosterol;Cholesterol 1.70E-05 1.42E-05 1.98E-05 1.94E-05 2.77E-05 2.79E-05 LPE(16:0);1-Palmitoyl-glycero-3-phosphoethanolamine 8.22E-06 N-Hexanoylsphingosine 1.83E-05 1.32E-05 3.89E-05 Oleamide 0.050601911 0.065527148 0.062965145 0.065565919 0.066602967 0.07870469 Sitosterol 0.012438472 0.009360131 0.009537235 0.016306016 0.014177281 0.010063017 0.012617407 0.013021139 0.011778704 Stigmasterol 0.001256205 0.001046652 0.000985259 0.001619542 0.001457835 0.001108639 0.001365757 0.001551718 0.001588329 Testosterone acetate 2.54E-05 5.98E-06 γ-Tocopherol 0.000683796 0.000510056 0.000556173 0.000844467 0.000829542 0.00053372 0.000648828 0.000866872 0.000814924 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PubChem CID m/z MT/RT 1,2-Distearoyl-glycero-3-phosphocholine 94190 790.6131167 17.15315967 15-Deoxy-Δ12,14-prostaglandin J2 5311211 317.2111617 11.82126458 17α-Estradiol 68570 273.175 9.842343 17α-Hydroxyprogesterone 6238 331.22582 10.05158067 1-Deoxysphinganine 9925886 286.30935 11.47844525 1-Oleoyl-glycero-3-phosphocholine 16081932 522.3554633 13.22100467 1-Palmitoyl-glycero-3-phosphocholine 460602 496.33903 13.07660067 1-Stearoyl-glycero-3-phosphocholine 497299 524.3709533 13.81711367 22-Hydroxycholesterol 167685 385.3404 13.272583 3β-Hydroxy-5-cholestenoic acid 165511 399.335695 13.4448075 5α-Cholestan-3-one 92128 387.36074 16.31990467 6-Gingerol 442793 277.178565 9.96433025 7-Dehydrocholesterol 439423 367.3364867 15.72026733 Abietic acid 10569 303.232065 13.706228 Acylcarnitine(18:1) 6426855 426.3492075 11.68139025 Acylcarnitine(20:1) 71464507 454.3780383 12.42848267 AEA(18:3) 5283449 322.27429 12.839632 AEA(20:3) 5283447 350.3046367 13.60236667 AEA(22:6) 5283451 372.29706 13.217424 Anandamide 5281969 348.29153 13.18397883 Bisacurone 14287397 217.1576075 8.3768085 Campesterol 173183 383.3679167 16.120143 Coenzyme Q10 5281915 863.6891433 18.726759 Dehydrozingerone 5354238 193.0865567 7.180045517 Deoxycorticosterone 6166 331.23376 9.928236 Desmosterol 439577 367.3298333 15.54254183 Eleutheroside B 5316860 390.1757333 5.515921 Etiocholan-3α-ol-17-one 5880 308.25497 11.039667 Flavanone 10251 225.09076 10.617534 Glabridin 124052 325.14336 11.30676667 Glucosylceramide(d18:1/24:1) 20057357 810.6655833 17.31222317 Hecogenin 91453 431.313645 12.3512295 Hydroxyprogesterone caproate 169870 429.2978233 12.17671283 Indole-3-carboxaldehyde 10256 146.0601233 6.8895791 Isofraxidin 5318565 223.0600633 6.4267569 Kynurenic acid 3845 190.04944 5.929567 Linoleyl ethanolamide 5283446 324.2897933 13.29135033 Lutein 5281243 551.4159633 14.78718833 N-Oleoylglycine 6436908 340.2854633 14.1039185 PAF(17:0) 24779315 538.39005 13.83562867 Palmitoylcarnitine 11953816 400.343735 11.518237 Palmitoylcholine 151731 342.3372617 11.408423 Palmitoylethanolamide 4671 300.29009 13.61718267 Phytosphingosine 122121 318.29982 10.88836833 Prostaglandin B2 5280881 335.21686 10.617534 Retinol 445354 269.22699 13.817209 Riboflavin 493570 377.149075 5.9700087 Sphinganine 91486 302.30696 11.179474 Sphinganine(d20:0) 137700203 330.3370333 11.725157 Sphinganine(d22:0) 358.367914 12.5438068 Sphingomyelin(d18:1/16:0) 9939941 703.5691133 16.20145867 Sphingomyelin(d18:1/18:0) 6453725 731.619828 16.4983954 Sphingosine 5280335 300.29004 10.995234 Sphingosine(d20:1) 328.3193367 11.54372933 Stearoyl ethanolamide 27902 328.32126 14.32807133 Trilaurin 10851 656.59565 17.34939967 Triptolide 107985 361.16459 7.407941333 Tropic acid 10726 167.0711167 5.646850567 Zeaxanthin 5280899 569.4335333 14.79829967 α-Tocopherol 14985 431.3855467 16.12384767 α-Tocopherol acetate 86472 490.427 16.37192133 β-Cryptoxanthin 5281235 553.4356267 16.397831 β-Estradiol 5757 273.18702 9.822026333 2-Aminoadipic acid;O-Acetylhomoserine;Glutamic acid γ-methyl ester 439389 385.3404 13.272583 6-Keto-prostaglandin E1 5280889 277.178565 9.96433025 9(S)-HODE 5312830 279.23223 12.47291617 AEA(18:1);Oleoyl ethanolamide 5283454 326.3055733 13.78009267 AEA(22:4) 5282273 376.318 13.75 Ethyl arachidonate 5367369 333.2780433 14.802005 Lanosterol;Cycloartenol 92110 409.3827433 16.097927 Lathosterol;Cholesterol 5997 369.35374 16.14961033 LPE(16:0);1-Palmitoyl-glycero-3-phosphoethanolamine 58177710 454.29016 13.005967 N-Hexanoylsphingosine 5702613 380.3504 14.295207 Oleamide 5283387 282.2795783 14.20019283 Sitosterol 222284 397.3757933 16.13865533 Stigmasterol 5280794 395.3673267 16.134957 Testosterone acetate 92145 331.21857 11.317416 γ-Tocopherol 92729 417.3695167 15.93871033 METABOLITES_END #END