#METABOLOMICS WORKBENCH n_hiraoka_20240509_004912 DATATRACK_ID:4828 STUDY_ID:ST003298 ANALYSIS_ID:AN005405 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 NEGATIVE 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 10-Hydroxyoctadecanoic acid 1.39E-05 7.29E-06 1.95E-05 3.70E-05 3.10E-05 5.42E-05 2-Hydroxydecanoic acid 4.15E-06 2-Hydroxyhexadecanoic acid 4.80E-06 4.22E-05 7.33E-05 8.54E-06 6.12E-06 8.90E-06 3-Hydroxydodecanoic acid 1.96E-05 3-Hydroxyhexadecanoic acid 3.55E-05 5.06E-05 9.80E-05 4.64E-05 4.47E-05 5.05E-05 3-Hydroxytetradecanoic acid 6.24E-05 5.81E-05 6.01E-05 6.18E-05 8.05E-05 6.50E-05 6.24E-05 6.69E-05 5.09E-05 Arachidic acid 0.001781409 0.00084946 0.000707195 0.000735542 0.001114372 0.000690914 0.000706726 0.001060317 0.00073633 Arachidonic acid 1.48E-06 9.53E-06 1.37E-05 1.64E-05 1.38E-05 Behenic acid 0.000271139 0.000140893 0.000129 0.000119881 0.000187464 0.000113377 0.000123385 0.000189949 0.000130376 Cholesterol sulfate 1.94E-05 9.02E-06 1.07E-05 1.06E-05 cis-11,14-Eicosadienoic acid 0.000233733 0.000105201 0.000125009 9.48E-05 0.000163501 0.000113212 0.000149629 0.000145089 0.00013244 cis-11-Eicosenoic acid 0.002777317 0.001102261 0.001274977 0.0009772 0.001547656 0.000893065 0.001263132 0.001504099 0.001379039 cis-5,8,11,14,17-Eicosapentaenoic acid 3.98E-06 5.91E-06 8.25E-06 1.30E-05 4.08E-06 6.95E-06 5.88E-06 9.59E-06 cis-8,11,14-Eicosatrienoic acid 3.25E-05 1.82E-05 3.23E-05 2.45E-05 3.44E-05 2.35E-05 2.34E-05 2.27E-05 2.51E-05 Docosanedioic acid 2.63E-05 1.98E-05 0.000116819 2.12E-05 2.64E-05 2.23E-05 Erucic acid 1.51E-05 1.05E-05 1.20E-05 1.91E-05 9.95E-06 7.16E-06 3.34E-05 2.35E-05 3.53E-05 Fatty acid(12:0) 1.14E-05 8.15E-06 1.74E-05 1.82E-05 2.53E-05 1.35E-05 1.84E-05 2.47E-05 2.20E-05 Fatty acid(14:1) 0.000105626 4.61E-05 4.86E-05 4.56E-05 6.23E-05 6.02E-05 4.72E-05 6.48E-05 3.39E-05 Fatty acid(14:2) 0.000318749 0.000205844 0.000186609 0.000289833 0.000326371 0.000277267 0.000232843 0.000348517 0.000216234 Fatty acid(14:3) 0.001007924 0.001060189 0.000558782 0.001634888 0.000461436 0.000468262 0.001409191 0.001272265 0.000758604 Fatty acid(15:1) 1.23E-05 8.60E-06 6.13E-06 7.73E-06 7.61E-06 7.28E-06 6.35E-06 6.35E-06 8.01E-06 Fatty acid(16:2) 0.000357916 0.000224708 0.00026199 0.00030533 0.000379947 0.000244769 0.000302248 0.000321111 0.000314234 Fatty acid(17:1) 9.78E-05 4.38E-05 7.01E-05 4.11E-05 6.64E-05 4.77E-05 5.71E-05 6.16E-05 5.14E-05 Fatty acid(17:2) 4.43E-05 2.96E-05 3.15E-05 2.84E-05 4.95E-05 2.55E-05 4.15E-05 3.64E-05 3.95E-05 Fatty acid(17:3) 8.77E-06 1.26E-05 Fatty acid(19:0) 9.55E-05 3.67E-05 3.41E-05 2.74E-05 4.78E-05 2.33E-05 3.17E-05 4.67E-05 3.29E-05 Glycochenodeoxycholic acid 2.03E-06 Hexadecanedioic acid 9.98E-06 1.44E-05 6.00E-05 7.75E-06 8.46E-06 1.26E-05 Hyodeoxycholic acid 3.13E-06 Lignoceric acid 1.12E-05 1.43E-05 5.85E-06 1.28E-05 1.61E-05 1.02E-05 Linoleic acid 0.003511242 0.00211301 0.002382549 0.001809267 0.002499306 0.001782886 0.002333319 0.002182989 0.002289736 LPG(16:0) 1.32E-05 4.81E-06 5.38E-06 Myristic acid 0.000867402 0.000586711 0.000727253 0.000842249 0.001003338 0.000581875 0.0006548 0.000769587 0.00086072 Octadecanedioic acid 1.15E-05 9.48E-06 4.56E-05 5.70E-06 9.31E-06 8.78E-06 Oleic acid 0.012268325 0.005283942 0.005152264 0.003408934 0.006266679 0.004589328 0.004961596 0.005024989 0.003986457 Palmitic acid 0.017204857 0.009659666 0.011932264 0.009420027 0.011004013 0.007836773 0.009528842 0.011099948 0.012060823 Palmitoleic acid 0.002473873 0.001061877 0.001208804 0.001010035 0.001668673 0.001124285 0.00136201 0.001368857 0.001257054 Pentadecanoic acid 9.06E-06 1.33E-05 1.13E-05 1.23E-05 8.08E-06 9.95E-06 1.19E-05 9.65E-06 5.97E-06 Ricinoleic acid 8.45E-05 7.98E-05 0.000101549 5.43E-05 0.000114889 5.87E-05 8.91E-05 0.000122096 7.73E-05 Stearic acid 0.012753024 0.006375978 0.007237121 0.006312485 0.007176862 0.004323092 0.004851709 0.00706525 0.006210497 Stearidonic acid 1.73E-05 1.33E-05 6.77E-06 2.14E-05 2.65E-05 4.88E-06 1.18E-05 1.78E-05 Tetradecanedioic acid 7.54E-06 1.07E-05 5.94E-05 7.13E-06 8.72E-06 Tricosanoic acid 2.67E-05 8.49E-06 1.36E-05 5.69E-06 1.10E-05 8.80E-06 8.27E-06 1.35E-05 7.16E-06 16-Hydroxyhexadecanoic acid 4.03E-05 2.84E-05 3.52E-05 1.85E-05 3.52E-05 3.34E-05 19-Methylarachidic acid;Heneicosanoic acid 3.34E-05 1.76E-05 1.58E-05 1.42E-05 2.55E-05 2.34E-05 1.92E-05 2.88E-05 1.56E-05 2-Hydroxyoctadecanoic acid 9.78E-06 0.00011129 0.000195784 1.15E-05 1.17E-05 5.51E-06 2-Hydroxytetradecanoic acid 2.02E-05 6.47E-06 cis-10-Nonadecenoic acid 2.88E-05 4.57E-05 2.71E-05 4.36E-05 3.77E-05 3.53E-05 Fatty acid(16:3) 2.72E-05 2.02E-05 2.31E-05 1.00E-05 2.20E-05 2.04E-05 Fatty acid(19:1) 7.27E-05 2.62E-05 3.28E-05 Fatty acid(19:2) 0.000119177 4.39E-05 6.00E-05 3.53E-05 5.30E-05 4.12E-05 6.52E-05 5.43E-05 6.21E-05 Fatty acid(24:0) 1.56E-05 9.31E-06 5.70E-06 Linolenic acid;γ-Linolenic acid 0.006075953 0.004517524 0.006301781 0.005185934 0.005733606 0.003322791 0.00417782 0.004279452 0.004733064 Taurolithocholic acid 3-sulfate 8.47E-05 6.82E-05 5.86E-05 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PubChem CID m/z MT/RT 10-Hydroxyoctadecanoic acid 9561835 299.2595583 13.26419333 2-Hydroxydecanoic acid 21488 187.1321 11.110066 2-Hydroxyhexadecanoic acid 92836 271.2260667 13.96001883 3-Hydroxydodecanoic acid 94216 215.16344 11.776517 3-Hydroxyhexadecanoic acid 301590 271.2274033 13.60471017 3-Hydroxytetradecanoic acid 16064 243.19697 12.58062033 Arachidic acid 10467 311.29601 15.623949 Arachidonic acid 444899 303.23434 14.040591 Behenic acid 8215 339.3270333 16.09045433 Cholesterol sulfate 65076 465.3037833 16.475493 cis-11,14-Eicosadienoic acid 6439848 307.2644233 14.70207233 cis-11-Eicosenoic acid 5282768 309.2800833 15.11302783 cis-5,8,11,14,17-Eicosapentaenoic acid 446284 301.21904 13.67208 cis-8,11,14-Eicosatrienoic acid 5280581 305.24882 14.31333067 Docosanedioic acid 244872 369.30129 14.28202458 Erucic acid 5281116 337.3110333 15.63506233 Fatty acid(12:0) 3893 199.16973 13.11799967 Fatty acid(14:1) 5281119 225.1859617 13.48256683 Fatty acid(14:2) 165415616 223.1705633 12.95144175 Fatty acid(14:3) 0 221.1550417 12.45177967 Fatty acid(15:1) 11708534 239.202705 13.8822995 Fatty acid(16:2) 5312427 251.202095 13.64542475 Fatty acid(17:1) 5282748 267.23326 14.45598725 Fatty acid(17:2) 6431005 265.2170433 13.97297592 Fatty acid(17:3) 134728675 263.201275 13.460308 Fatty acid(19:0) 12591 297.2793433 15.54783833 Glycochenodeoxycholic acid 12544 448.3104 11.210706 Hexadecanedioic acid 10459 285.2080067 12.14087767 Hyodeoxycholic acid 5283820 391.28525 11.213158 Lignoceric acid 11197 367.35978 16.70262233 Linoleic acid 5280450 281.23968 14.098591 LPG(16:0) 3300276 483.2717533 15.07230033 Myristic acid 11005 227.2018133 13.754272 Octadecanedioic acid 70095 313.2384767 12.938489 Oleic acid 445639 282.25222 14.5576855 Palmitic acid 985 255.23309 14.46512567 Palmitoleic acid 445638 253.21746 13.93569067 Pentadecanoic acid 13849 241.2181267 14.131907 Ricinoleic acid 643684 297.2444667 12.51398733 Stearic acid 5281 284.26789 15.079704 Stearidonic acid 5312508 275.2015133 13.29148133 Tetradecanedioic acid 13185 257.177404 11.2654008 Tricosanoic acid 17085 353.3425833 16.28667633 16-Hydroxyhexadecanoic acid 10466 271.2281367 12.29447667 19-Methylarachidic acid;Heneicosanoic acid 3632876 325.3113833 16.04750533 2-Hydroxyoctadecanoic acid 69417 299.25665 14.765005 2-Hydroxytetradecanoic acid 1563 243.198895 12.982825 cis-10-Nonadecenoic acid 5312513 295.25998 14.942233 Fatty acid(16:3) 23019726 249.1872 13.1827675 Fatty acid(19:1) 52921844 295.26486 14.86497467 Fatty acid(19:2) 134746205 293.24815 14.5650825 Fatty acid(24:0) 10467 367.35772 16.505111 Linolenic acid;γ-Linolenic acid 5280934 278.22083 13.88229867 Taurolithocholic acid 3-sulfate 440071 562.2608667 13.698734 METABOLITES_END #END