#METABOLOMICS WORKBENCH SIMR_Core_Facility_20240430_224506 DATATRACK_ID:4804 STUDY_ID:ST003194 ANALYSIS_ID:AN005242 PROJECT_ID:PR001990 VERSION 1 CREATED_ON May 5, 2024, 8:50 pm #PROJECT PR:PROJECT_TITLE Multi-Omics Analysis Revealed Significant Metabolic Changes in Brain Cancer PR:PROJECT_TITLE Cells Treated with Paclitaxel and/or Topotecan PR:PROJECT_SUMMARY Glioblastoma (GB) is the most common primary malignant brain tumor, representing PR:PROJECT_SUMMARY approximately 57% of all gliomas and 48% of all primary malignant central PR:PROJECT_SUMMARY nervous system (CNS) tumors. Despite the best standard therapies, glioblastoma PR:PROJECT_SUMMARY survivors have a brief survival time, about 24 months on average. The treatment PR:PROJECT_SUMMARY is troublesome because the cancer cells may not respond well to specific PR:PROJECT_SUMMARY therapies as they grow within an extensive network of blood vessels. A PR:PROJECT_SUMMARY multi-omics approach to provide information on biomolecules from multiple layers PR:PROJECT_SUMMARY appears promising for systematically and holistically understanding complex PR:PROJECT_SUMMARY biology. However, till now, only few studies have utilized omics analysis to PR:PROJECT_SUMMARY investigate the impact of anticancer drugs on GBM. Our study aimed to evaluate PR:PROJECT_SUMMARY the impacts of the anticancer medications (paclitaxel 5.3 μg/mL and topotecan PR:PROJECT_SUMMARY 0.26 μM) solely and in pairwise combination on the metabolic and proteomic PR:PROJECT_SUMMARY signatures of the U87 cell line while utilizing the accurate PR:PROJECT_SUMMARY ultra-high-performance liquid chromatography-electrospray ionization quadrupole PR:PROJECT_SUMMARY time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) analytical technology. The PR:PROJECT_SUMMARY studied cancer cells wear treated with DMSO (control group), paclitaxel 5.3 µM, PR:PROJECT_SUMMARY topotecan 0.26 µM, and a combination of paclitaxel 5.3 µM and topotecan 0.26 PR:PROJECT_SUMMARY µM. Using One-way ANOVA, we observed 14 significantly altered metabolites PR:PROJECT_SUMMARY compared to those cells treated with DMSO. For combination treatment (paclitaxel PR:PROJECT_SUMMARY and topotecan), 10 metabolites were significantly dysregulated. The sparse PR:PROJECT_SUMMARY partial least squares-discriminant analysis (sPLS-DA) showed minimal PR:PROJECT_SUMMARY overlapping, indicating a difference between the four groups. While for PR:PROJECT_SUMMARY proteomics, a total of 79 proteins were significantly dysregulated among the PR:PROJECT_SUMMARY groups. These findings can aid in identifying new biomarkers associated with the PR:PROJECT_SUMMARY utilized drugs and creating a map for targeted therapy. EIF3F, GNB2L1, HINT2, PR:PROJECT_SUMMARY and RPA3 were shown to be significantly upregulated in the combination group PR:PROJECT_SUMMARY when compared to the control. Moreover, ribosome, apoptosis, HIF-1 signaling, PR:PROJECT_SUMMARY arginine and proline, glutathione, purine metabolism, apelin signaling pathway, PR:PROJECT_SUMMARY and glycolysis were significantly altered in the combination group PR:INSTITUTE Sharjah Institute for Medical Research PR:LAST_NAME Facility PR:FIRST_NAME Core PR:ADDRESS M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, PR:ADDRESS UAE, Sharjah, 000, United Arab Emirates PR:EMAIL tims-tof@sharjah.ac.ae PR:PHONE +971 6 5057656 #STUDY ST:STUDY_TITLE Multi-Omics Analysis Revealed Significant Metabolic Changes in Brain Cancer ST:STUDY_TITLE Cells Treated with Paclitaxel and/or Topotecan ST:STUDY_SUMMARY Glioblastoma (GB) is the most common primary malignant brain tumor, representing ST:STUDY_SUMMARY approximately 57% of all gliomas and 48% of all primary malignant central ST:STUDY_SUMMARY nervous system (CNS) tumors. Despite the best standard therapies, glioblastoma ST:STUDY_SUMMARY survivors have a brief survival time, about 24 months on average. The treatment ST:STUDY_SUMMARY is troublesome because the cancer cells may not respond well to specific ST:STUDY_SUMMARY therapies as they grow within an extensive network of blood vessels. A ST:STUDY_SUMMARY multi-omics approach to provide information on biomolecules from multiple layers ST:STUDY_SUMMARY appears promising for systematically and holistically understanding complex ST:STUDY_SUMMARY biology. However, till now, only few studies have utilized omics analysis to ST:STUDY_SUMMARY investigate the impact of anticancer drugs on GBM. Our study aimed to evaluate ST:STUDY_SUMMARY the impacts of the anticancer medications (paclitaxel 5.3 μg/mL and topotecan ST:STUDY_SUMMARY 0.26 μM) solely and in pairwise combination on the metabolic and proteomic ST:STUDY_SUMMARY signatures of the U87 cell line while utilizing the accurate ST:STUDY_SUMMARY ultra-high-performance liquid chromatography-electrospray ionization quadrupole ST:STUDY_SUMMARY time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) analytical technology. The ST:STUDY_SUMMARY studied cancer cells wear treated with DMSO (control group), paclitaxel 5.3 µM, ST:STUDY_SUMMARY topotecan 0.26 µM, and a combination of paclitaxel 5.3 µM and topotecan 0.26 ST:STUDY_SUMMARY µM. Using One-way ANOVA, we observed 14 significantly altered metabolites ST:STUDY_SUMMARY compared to those cells treated with DMSO. For combination treatment (paclitaxel ST:STUDY_SUMMARY and topotecan), 10 metabolites were significantly dysregulated. The sparse ST:STUDY_SUMMARY partial least squares-discriminant analysis (sPLS-DA) showed minimal ST:STUDY_SUMMARY overlapping, indicating a difference between the four groups. While for ST:STUDY_SUMMARY proteomics, a total of 79 proteins were significantly dysregulated among the ST:STUDY_SUMMARY groups. These findings can aid in identifying new biomarkers associated with the ST:STUDY_SUMMARY utilized drugs and creating a map for targeted therapy. EIF3F, GNB2L1, HINT2, ST:STUDY_SUMMARY and RPA3 were shown to be significantly upregulated in the combination group ST:STUDY_SUMMARY when compared to the control. Moreover, ribosome, apoptosis, HIF-1 signaling, ST:STUDY_SUMMARY arginine and proline, glutathione, purine metabolism, apelin signaling pathway, ST:STUDY_SUMMARY and glycolysis were significantly altered in the combination group. ST:INSTITUTE Sharjah Institute for Medical Research ST:LAST_NAME Facility ST:FIRST_NAME Core ST:ADDRESS M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, ST:ADDRESS UAE, Sharjah, 000, United Arab Emirates ST:EMAIL tims-tof@sharjah.ac.ae ST:PHONE +971 6 5057656 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Brain Cancer Cells #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 - U87 A1-01-4711 Treatment:DMSO | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A1-01-4711.d SUBJECT_SAMPLE_FACTORS - U87 A1-02-4712 Treatment:DMSO | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A1-02-4712.d SUBJECT_SAMPLE_FACTORS - U87 B1-01-4713 Treatment:DMSO | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B1-01-4713.d SUBJECT_SAMPLE_FACTORS - U87 B1-02-4714 Treatment:DMSO | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B1-02-4714.d SUBJECT_SAMPLE_FACTORS - U87 A2-01-4715 Treatment:Paclitaxel | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A2-01-4715.d SUBJECT_SAMPLE_FACTORS - U87 A2-02-4716 Treatment:Paclitaxel | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A2-02-4716.d SUBJECT_SAMPLE_FACTORS - U87 B2-01-4717 Treatment:Paclitaxel | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B2-01-4717.d SUBJECT_SAMPLE_FACTORS - U87 B2-02-4718 Treatment:Paclitaxel | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B2-02-4718.d SUBJECT_SAMPLE_FACTORS - U87 A3-01-4719 Treatment:Etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A3-01-4719.d SUBJECT_SAMPLE_FACTORS - U87 A3-02-4720 Treatment:Etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A3-02-4720.d SUBJECT_SAMPLE_FACTORS - U87 B3-01-4721 Treatment:Etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B3-01-4721.d SUBJECT_SAMPLE_FACTORS - U87 B3-02-4722 Treatment:Etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B3-02-4722.d SUBJECT_SAMPLE_FACTORS - U87 A4-01-4723 Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A4-01-4723.d SUBJECT_SAMPLE_FACTORS - U87 A4-02-4724 Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 A4-02-4724.d SUBJECT_SAMPLE_FACTORS - U87 B4-01-4725 Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B4-01-4725.d SUBJECT_SAMPLE_FACTORS - U87 B4-02-4726 Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line RAW_FILE_NAME(Raw file name)=U87 B4-02-4726.d #COLLECTION CO:COLLECTION_SUMMARY Since the U87 cell line is one of the most frequently used and thoroughly CO:COLLECTION_SUMMARY defined brain cancer cell lines, it was chosen as the in-vitro model for this CO:COLLECTION_SUMMARY study. This investigation used the DMEM medium supplemented with 10% fetal CO:COLLECTION_SUMMARY bovine serum and 1% penicillin/streptomycin to cultivate the U87 cell line as CO:COLLECTION_SUMMARY monolayers Sigma-Aldrich (Louis, USA). All cultures were grown at 37 °C in a CO:COLLECTION_SUMMARY humid environment with 5% CO2. CO:SAMPLE_TYPE Cultured brain cancer cells #TREATMENT TR:TREATMENT_SUMMARY For each treatment condition, every 75 cm2 tissue culture flask contained two TR:TREATMENT_SUMMARY million cells, which were then incubated for 24 hours. The cells were TR:TREATMENT_SUMMARY subsequently given a 24-hour treatment of paclitaxel 5.3 μM and/or topotecan TR:TREATMENT_SUMMARY 0.26 μM. According to cytotoxicity assay results, these doses correspond to the TR:TREATMENT_SUMMARY IC50 of these substances for the U87 cell line. The control group was Dimethyl TR:TREATMENT_SUMMARY sulfoxide (DMSO) at a 0.5% concentration for 24 hours. After the incubation TR:TREATMENT_SUMMARY period, cells were washed twice with phosphate-buffered saline solution (PBS) TR:TREATMENT_SUMMARY then collected by trypsinization and resuspended in 1 mL of the media for TR:TREATMENT_SUMMARY further analysis. Cells were then separated into pellets by centrifugation for 5 TR:TREATMENT_SUMMARY minutes at room temperature at 1300 rpm. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY First, one tablet of protease inhibitor was prepared in 10 mL lysis buffer, and SP:SAMPLEPREP_SUMMARY 400 μL of the mixture was added to each sample. Then the samples were rested SP:SAMPLEPREP_SUMMARY for 10 minutes and transferred into 10 mL tubes, followed by vortex for 2 SP:SAMPLEPREP_SUMMARY minutes and sonication using a COPLEY probe-sonicator (QSONICA SONICATOR, USA) SP:SAMPLEPREP_SUMMARY for 30 seconds while utilizing a 30% amplifier in an ice bath. Next, the samples SP:SAMPLEPREP_SUMMARY were transferred to Eppendorf tubes and centrifuged at 14000 rpm for 5 minutes. SP:SAMPLEPREP_SUMMARY Next, the supernatant was transferred to the new Eppendorf, and 400 μL of SP:SAMPLEPREP_SUMMARY methanol and 300 μL of chloroform were added to the supernatant. Then we SP:SAMPLEPREP_SUMMARY vortexed the samples for 30 seconds and centrifuged them at 14000 rpm for 5 SP:SAMPLEPREP_SUMMARY minutes. Afterward, a white disk appeared. Finally, we collected the upper layer SP:SAMPLEPREP_SUMMARY into a new glass vial, added 300 μL of methanol to the bottom layer and white SP:SAMPLEPREP_SUMMARY disk, and vortexed until the white disk was broken and precipitated. Then we SP:SAMPLEPREP_SUMMARY centrifuged the samples at 14000 rpm for 3 minutes and transferred the SP:SAMPLEPREP_SUMMARY supernatant to the previously prepared glass vials for metabolomics analysis SP:SAMPLEPREP_SUMMARY using UHPLC-QTOF-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Elute UHPLC, Hamilton® Intensity Solo 2 C18 column (100 mm × 2.1 mm, 1.8 m CH:CHROMATOGRAPHY_SUMMARY beads), autosampler (Elute UHPLC), solvent delivery pump (HPG 1300) were CH:CHROMATOGRAPHY_SUMMARY utilized to separate and detect the metabolites and peptides using inline CH:CHROMATOGRAPHY_SUMMARY reversed-phase chromatography with solvent A 0.1% formic acid in HPLC grade CH:CHROMATOGRAPHY_SUMMARY water and solvent B 0.1% formic acid in ACN. in the cells (Bruker, Bremen, CH:CHROMATOGRAPHY_SUMMARY Germany. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Bruker Elute CH:COLUMN_NAME Bruker Hamilton Intensity Solo 2 C18(100 mm × 2.1 mm, 1.8 m beads) CH:SOLVENT_A 100% Water; 0.1% Formic Acid CH:SOLVENT_B 100% Acetonitrile; 0.1% Formic Acid CH:FLOW_GRADIENT The metabolomics samples were injected twice and eluted using a 30-min gradient CH:FLOW_GRADIENT as follows: 1% ACN for 2 minutes, ramping to 99% ACN for 15 minutes, retained at CH:FLOW_GRADIENT 99% ACN for 3 minutes, then re-equilibration to 1% ACN for ten minutes CH:FLOW_RATE For elution, the flow rate was 250 L/min, and for re-equilibration, it was 350 CH:FLOW_RATE L/min. A 10 L/min flow of drying gas CH:COLUMN_TEMPERATURE 35 °C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker timsTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The data was processed using MetaboScape® 4.0 software (Bruker, Bremen, MS:MS_COMMENTS Germany) [19]. In the T-ReX 2D/3D workflow, the following settings were applied MS:MS_COMMENTS to molecular feature detection: a minimum intensity threshold of 1,000 counts, a MS:MS_COMMENTS maximum peak duration of seven spectra, and a maximum peak area. The mass MS:MS_COMMENTS recalibration was completed within a retention time range of 0 to 0.3 minutes. MS:MS_COMMENTS Only features found in at six four of the 24 samples (per cell type) were MS:MS_COMMENTS considered. Similarly, the MS/MS import method was set to be averaged. MS:MS_COMMENTS Accordingly, we assigned the following bucketing parameters to the data: MS:MS_COMMENTS retention duration of 0.3 to 25 minutes and mass range of 50 to 1,000 m/z. For MS:MS_COMMENTS accurate identification, the following standards were set up: Initially unknown MS:MS_COMMENTS compounds from QTOF MS data were characterized using MS/MS spectra and retention MS:MS_COMMENTS time (RT). The Human Metabolome Database (HMDB) 4.0, a database of annotated MS:MS_COMMENTS metabolomics resources, spectrum library, was used to annotate compounds that MS:MS_COMMENTS made it past the screening and exhibited MS/MS or MS/MS in conjunction with RT MS:MS_COMMENTS [20]. This collection served as a benchmark against which all finally chosen MS:MS_COMMENTS chemicals were evaluated. To determine which metabolites were present, we mapped MS:MS_COMMENTS MS/MS spectra and retention durations using the HMDB 4.0. When multiple features MS:MS_COMMENTS matched a single database entry, the metabolites were filtered by selecting the MS:MS_COMMENTS entry of each metabolite with the highest annotation quality score (AQ score), MS:MS_COMMENTS i.e. the best fit with the most factors including retention time, MS/MS, m/z MS:MS_COMMENTS values, analyte list, msigma, and spectral library. Furthermore, for analysis, MS:MS_COMMENTS metabolite data was saved as CSV files and integrated into the complete MS:MS_COMMENTS metabolomics platform MetaboAnalyst 5.0 software (https://www.metaboanalyst.ca) MS:MS_COMMENTS [21]. For each medication, two-tailed independent student t-tests were employed MS:MS_COMMENTS to identify significantly different metabolites from DMSO. For each condition, a MS:MS_COMMENTS volcano plot illustrating statistical significance and fold change for cellular MS:MS_COMMENTS metabolite dysregulation was created. Through the use of a one-way analysis of MS:MS_COMMENTS variance, many groups were compared (ANOVA). The threshold for significance was MS:MS_COMMENTS fixed at p <0.05. Using the software MetaboAnalyst 5.0, the Principal Component MS:MS_COMMENTS Analysis (PCA) was also performed to compare the two groups. The false discovery MS:MS_COMMENTS rate was used to eliminate false positives and fix multiple hypothesis testing MS:MS_COMMENTS (FDR). Enrichment analysis, joint pathway analysis, and heatmaps were also MS:MS_COMMENTS created using MetaboAnalyst. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AU MS_METABOLITE_DATA_START Samples U87 A1-01-4711 U87 A1-02-4712 U87 B1-01-4713 U87 B1-02-4714 U87 A2-01-4715 U87 A2-02-4716 U87 B2-01-4717 U87 B2-02-4718 U87 A3-01-4719 U87 A3-02-4720 U87 B3-01-4721 U87 B3-02-4722 U87 A4-01-4723 U87 A4-02-4724 U87 B4-01-4725 U87 B4-02-4726 Factors Treatment:DMSO | Sample source:Brain Cancer cell line Treatment:DMSO | Sample source:Brain Cancer cell line Treatment:DMSO | Sample source:Brain Cancer cell line Treatment:DMSO | Sample source:Brain Cancer cell line Treatment:Paclitaxel | Sample source:Brain Cancer cell line Treatment:Paclitaxel | Sample source:Brain Cancer cell line Treatment:Paclitaxel | Sample source:Brain Cancer cell line Treatment:Paclitaxel | Sample source:Brain Cancer cell line Treatment:Etoposide | Sample source:Brain Cancer cell line Treatment:Etoposide | Sample source:Brain Cancer cell line Treatment:Etoposide | Sample source:Brain Cancer cell line Treatment:Etoposide | Sample source:Brain Cancer cell line Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line Treatment:Paclitaxel and etoposide | Sample source:Brain Cancer cell line 1,3,5-Trimethoxybenzene 14794 14678 15456 14246 14016 16354 14088 21280 15398 16106 16554 15948 21604 21252 0 17960 2,4-Diaminobutyric acid 102130 95868 108028 106010 96408 93684 89606 93444 91798 86278 86606 85784 101450 90516 96818 82846 2-Hydroxy-3-methylbutyric acid 3400 3052 2582 2886 2966 2608 3016 2310 2548 2366 3282 2562 3514 2794 2550 2782 2-Hydroxycaproic acid 76584 77820 78576 74908 75572 74818 71436 68752 73214 75010 75178 75600 78006 79982 77174 77534 2-Phenylbutyric acid 33938 30814 30328 29144 33446 28288 27808 26972 30482 28526 29154 30742 32838 32964 31020 31440 2-Pyrrolidinone 7072 9132 7844 7226 8246 8628 8782 7888 7826 7736 8502 8790 8610 7778 9120 8292 3,4,5-Trimethoxycinnamic acid 1568 2234 1946 1648 1430 1984 2070 1896 1412 1664 1886 2020 1920 1974 1806 1548 3,4-Dihydroxybenzeneacetic acid 3328 3740 3472 3456 3618 3768 3032 3192 3226 3420 4610 3626 2988 3134 3286 2886 3,4-Dihydroxyphenylglycol 2374 2702 2230 2558 2438 2018 2118 2242 2586 2428 2228 2284 2102 2324 2374 2382 3-Aminoisobutanoic acid 575006 581006 541308 560526 558868 540186 531904 529254 526742 544892 570106 576186 563630 561490 564996 565224 3-Cresotinic acid 358 262 1760 2088 1456 1544 1030 1048 894 1208 1588 1390 1306 0 906 804 3-Hydroxyanthranilic acid 1346 4088 2322 4484 5636 6642 5110 5662 6480 4446 7262 234 0 1122 5306 0 3-Methyl-2-oxovaleric acid 2200 2310 2666 2100 1582 2318 2678 2638 1954 2052 2024 3002 2352 2396 2040 2702 3a,6b,7b-Trihydroxy-5b-cholanoic acid 8070 8526 6472 6200 7254 7440 7026 7924 8108 7258 8366 7178 6798 7228 7866 6028 4-Aminophenol 1584 1550 896 640 846 1140 1584 1762 5020 4912 1300 1104 886 960 1504 1584 4-Ethylbenzoic acid 2676 3854 3514 3748 2216 2004 2128 1658 4008 3526 3576 3672 2172 1870 2458 2586 4-Hydroxyproline 1194 1380 1800 2026 1900 1522 1978 1990 1748 1388 1628 1592 1880 2078 1652 1812 5'-Methylthioadenosine 1170 1126 1024 1266 1510 1246 1626 1620 1394 1122 1500 1336 986 854 1360 1404 5-Hydroxytryptophol 260 906 974 1000 108 310 822 694 1706 1730 2430 2850 2786 2250 834 1256 5-Methoxytryptophan 2456 2580 3032 2852 3088 2822 2306 3628 2592 2968 2742 3192 3338 2598 2684 3596 5-Methoxytryptophol 110 0 446 554 916 1252 260 412 790 1026 608 880 1568 2030 772 716 Acetaminophen 12378 10582 5724 6242 5754 5516 15044 15008 35500 42320 9118 8746 5040 6792 9364 9178 Adenine 1558 1848 1544 1466 248 92 70 0 1338 1012 1010 1318 306 122 356 98 Adipic acid 4148 4104 3884 4392 4320 3786 4398 4712 4694 4206 3858 4002 3732 3340 4444 4124 ADP 3366 3682 3786 3580 0 66 0 0 4160 3420 3442 3748 0 56 0 0 Androstenedione 11544 10834 8676 11970 7316 6332 7678 7426 10344 10080 10106 12154 9456 7686 8588 8398 Androsterone 26126 24902 25968 27140 27026 25602 26204 25824 26162 25110 25840 25272 26164 25154 27628 24974 Aspirin 18326 17364 19866 21692 20878 21396 12870 13234 18544 17754 19564 17420 18896 20778 11606 11282 Azelaic acid 2724 2314 2230 2756 2034 2120 4062 4984 3090 2410 2058 2202 1810 2118 1970 1908 Azythromycin 1138 1862 1562 1736 1572 1380 1198 1172 1132 1280 1144 1238 1504 1062 992 850 Benzaldehyde 13866 14022 14298 13242 15288 13830 10906 12320 11268 11862 12890 12724 16532 14036 13532 15364 Cinnamaldehyde 840 894 1064 1416 888 752 850 908 994 1102 1006 976 1106 930 778 820 Cortisol 54302 60128 61018 57574 57204 60302 59134 60460 59382 62950 59046 57336 56922 57694 56302 53286 Cyclic AMP 1102 1582 1324 1538 380 378 150 416 1694 1698 1168 672 198 232 188 104 Cytidine 1768 2308 2124 1358 0 0 0 0 1626 2056 1634 1468 0 0 0 0 Diacetyl 4332 3980 4278 3718 3920 3112 3650 3938 4614 3996 5404 5980 4060 4058 3520 4440 Dihydrothymine 408 258 1426 1300 1606 1904 640 878 1328 1248 1938 2528 1774 1920 1428 1434 Glycerol 18832 17592 19890 18856 19674 17634 18660 18640 20492 18564 18562 18920 18974 24122 17500 18844 Glycerophosphocholine 3440 3278 3184 3092 84 110 286 394 3196 3598 3158 3862 166 144 326 162 Glycyl-L-leucine 2302 2360 2182 1604 198 0 0 138 2798 2056 1836 1942 212 156 188 0 Guanine 7800 7060 6624 6926 156 140 208 0 9446 6184 7512 8502 94 178 106 102 Guanosine 6992 6362 6626 5530 128 294 0 0 4556 4722 5366 100 0 170 0 0 Guanosine 5'-diphosphate 3174 3412 2848 2418 0 0 0 0 3380 2554 3432 3620 0 0 0 0 Hydroquinone 1760 1986 1922 1904 1748 1626 2098 1882 1842 1864 2176 1602 1868 1806 1594 1688 Hypoxanthine 39912 45844 43236 41662 2116 2260 220 208 42156 43664 46976 29670 138 3474 600 810 Indoleacrylic acid 25686 24440 25554 23338 2766 2514 2884 2282 24652 24338 27090 25306 3836 2754 2978 2748 Inosine 4404 4530 3772 5186 414 328 212 510 4598 4072 5020 3312 170 570 154 344 Isobutyric acid 29952 29922 26102 34226 28316 29906 25980 19904 27884 30134 27206 30454 26246 32518 30830 32176 L(-)-Nicotine pestanal 1336 1422 1822 1588 3640 3878 1998 1822 1564 1872 1672 1396 3308 3538 2038 1942 L-Arginine 2934 3074 2904 2310 448 162 508 370 2344 2314 2256 2144 290 160 1048 1132 L-Dopa 234 1834 1558 1612 1478 1446 1310 1230 1616 1410 868 238 198 118 1124 680 L-Glutamic acid 14412 12440 13388 12658 13922 16000 14246 15138 12580 12732 12438 11612 11478 12524 12916 11236 L-Glutamine 1294 998 1228 934 2184 1640 1722 1388 1294 1816 1332 1294 0 1216 1006 1328 L-Histidine 1114 1314 1442 1674 1128 1180 624 796 1334 1340 1562 1876 1322 1524 692 476 L-Kynurenine 2140 2130 1976 1668 2488 2732 1848 1912 2128 2350 2394 1860 1788 2602 1950 2762 L-Methionine 5654 4154 3932 4616 0 98 124 332 1574 2966 4022 6166 648 676 574 742 L-Norleucine 13510 14038 11740 12416 9054 9832 9026 9022 12270 12166 12764 11904 11766 9956 10164 9708 L-Phenylalanine 28082 28482 24494 21794 2518 2142 2052 2016 26022 23156 20854 23402 2124 2524 3052 2834 L-Threonine 16982 16638 16104 15344 15042 15382 15954 15694 15812 15740 16146 15840 15682 15484 15210 16736 L-Tryptophan 5922 4342 4268 4354 688 772 744 812 5318 5360 5576 4818 698 612 920 866 Maltitol 165750 170344 161518 158814 165056 156650 152638 155666 161860 168208 160602 163912 161700 158774 162278 152812 Metoprolol 3594 3674 2404 2186 0 1428 2838 3598 1618 1810 1754 1964 1180 1178 2522 2310 Mevalonic acid 176458 189388 177384 174982 214120 201966 193884 195302 184268 188206 178640 166994 205648 211152 205528 221074 N,N-Dimethylformamide 4322 4350 4086 4540 4308 4646 3810 4404 4064 3196 4234 4394 3726 4628 4104 3894 N-Acetyl-L-alanine 13608 0 17124 16642 21968 19772 24728 19466 21572 18484 19612 20016 18976 18018 19288 19810 N-Acetyl-L-phenylalanine 1774 2042 1644 1914 1860 2044 2032 2518 2152 1896 2122 1834 2324 2044 1928 2242 N-Acetylputrescine 1250 920 1006 926 1086 1034 594 868 1022 964 804 608 1852 1220 830 1136 N-Acetylserotonin 3730 3716 4004 3870 4250 4410 3900 3760 4436 4472 3886 3730 3990 4364 3576 3558 Naproxen 1726 1396 1994 1634 1644 1562 1816 1390 1524 1564 2384 0 1356 1496 2558 2734 Niacinamide 16326 19440 18292 16904 2314 2214 646 826 17694 16392 15714 14784 2126 2256 1178 956 Norepinephrine 1802 2322 1436 1744 1640 1176 1174 1726 1750 1722 1460 1656 1590 2074 1402 2188 Nutriacholic acid 0 0 0 0 4470 3750 5446 5084 0 248 156 170 6582 4696 4504 3572 o-Tyrosine 19600 288 12876 14766 1290 1422 1328 1550 284 14828 21808 20798 2922 2990 2314 2990 p-Hydroxyphenylacetic acid 1972 1494 2194 1874 96 102 0 0 1116 1542 1772 0 0 0 0 0 p-Octopamine 1648 1862 1972 1320 1440 1428 1436 2112 1578 1510 1288 1604 1730 1376 1690 1740 PC(16:0/16:0) 172 0 68 74 2312 448 1010 3514 5562 4054 3948 3780 3472 1580 1200 996 PC(18:1(9Z)/18:1(9Z)) 696 672 0 646 2534 772 990 1286 1246 1104 1048 1188 328 320 342 174 Phenylacetaldehyde 23696 22210 23806 23540 23060 23096 22480 24914 23694 21962 21410 23038 24744 26336 23630 24438 Phenylacetic acid 2816 3792 3282 3506 3700 3538 2888 2894 3066 2954 3254 2962 3184 3504 3388 3020 Phenylacetylglycine 9292 9562 10202 9756 10464 10652 10090 10020 9740 9052 9798 10006 10474 9228 9394 10456 Phosphocreatine 82870 79038 79376 83112 82816 83994 76534 76352 86630 79478 80706 82948 87332 83586 89030 86622 Phosphoric acid 2410 2612 2644 2242 1892 2708 2488 3212 2364 2380 2174 2112 1972 1962 2026 2624 Pipecolic acid 3244 2728 3240 3138 3118 3018 2906 3152 3166 2782 2656 2154 3548 2818 2682 3068 Progesterone 10562 10252 9336 8718 9904 11836 10148 8968 10688 11296 10914 10750 10484 11656 10636 11664 Propionic acid 132630 136730 123494 125120 125858 141578 124622 127448 130898 129682 127232 123016 122470 124654 116454 140894 Pyroglutamic acid 1292 1558 2424 2246 254 166 124 0 870 1364 978 1176 134 236 260 190 Pyrrole-2-carboxylic acid 662 1098 1064 864 1204 744 1020 1078 862 1068 912 734 800 900 814 578 Rhamnose 1090 1114 1242 1662 862 1262 1170 1398 586 914 800 1224 802 1032 798 990 Riboflavin 772 548 968 872 840 1232 990 1074 754 998 964 836 1036 1192 776 862 Saccharopine 8758 7950 6660 6138 5420 5350 6782 7136 6958 5692 5458 5236 4932 5216 5394 5394 Salicin 2168 2036 2122 1704 2210 2320 2736 2936 2402 2018 2074 1728 1962 2506 2178 2140 Serotonin 13084 13576 14652 13290 13722 13814 13684 12584 14256 12804 13888 13374 14254 16346 14356 13508 Sorbitol 21088 22714 23318 23212 20784 22230 23336 23080 22156 22382 22428 21484 19890 19598 22012 20136 Spermine 17306 11448 10900 12082 2992 1524 880 506 7534 8582 13822 15282 5606 2010 1168 1056 Sphinganine 12868 13394 12160 13376 10874 9158 8508 7188 6694 6132 9468 8890 10900 10486 9394 12046 Succinylacetone 34956 32182 41626 41706 41512 44514 21746 24700 36696 34826 39660 37452 39174 44356 15524 18306 Sulfolithocholic acid 0 1516 0 1008 980 1262 1564 1368 1026 1418 1164 1312 1098 1070 1248 578 Taurocholic acid 1714 1434 1472 0 0 0 1808 0 970 1112 1268 0 1260 1468 1582 1394 Taurodeoxycholic acid 1548 1612 1644 1858 1772 1656 1426 1788 2152 1858 1914 2100 2628 1768 1368 1966 Tetrahydrocortisone 1026 1152 1174 0 1338 1310 1114 1054 1022 780 1066 1140 1346 1190 1282 1420 Thymidine 1976 1686 1016 1466 0 0 0 0 1820 1916 1520 1544 86 0 0 0 Tiglylglycine 1074 1608 1152 1012 872 982 1610 1176 1136 1254 1528 1396 928 1326 786 1474 Trehalose 968 1316 1114 1106 1150 1194 1156 160 1104 182 1202 1270 1066 920 1018 1130 Tryptophanol 2660 2404 1948 2540 2374 3202 2402 2172 2420 2514 2524 2084 2530 2922 2304 2750 Uracil 458 6384 5600 7476 300 290 258 224 5374 6052 100 108 0 196 248 400 Urea 78386 67932 74172 78238 74916 74086 68362 62888 57972 62252 56236 57230 68596 61170 48306 56576 Ureidosuccinic acid 5990 5232 6776 6346 6494 6600 5706 5064 5366 4946 6102 6352 6454 5442 5402 5514 Uric acid 31588 37646 34740 32778 3768 3758 4542 4176 34188 28134 31574 28582 3244 3236 0 3158 Uridine 24536 25298 26586 27224 24674 23880 23988 24984 22922 27272 24536 27112 25580 24158 20378 23450 Urocanic acid 3520 2804 2620 3094 1494 1672 1746 2716 1044 2200 2598 3154 3098 3948 2796 3274 Verapamil 1932 1504 1430 1768 1818 2016 1836 1654 1686 2114 1700 1614 2260 1940 2124 1834 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Molecular Formula RT [min] m/z meas. M meas. Ions AQ Boxplot ΔRT 1,3,5-Trimethoxybenzene C9H12O3 21.85 57.03367 168.07917 [M+H+H2]3+ 2100 15948 10.56 2,4-Diaminobutyric acid C4H10N2O2 14.83 119.08537 118.07809 [M+H]+ 1110 93684 13.92 2-Hydroxy-3-methylbutyric acid C5H10O3 13.73 119.07235 118.06507 [M+H]+ 1100 2794 8.16 2-Hydroxycaproic acid C6H12O3 16.22 133.08592 132.07864 [M+H]+ 2120 75600 9.32 2-Phenylbutyric acid C10H12O2 11.6 165.09106 164.08379 [M+H]+ 2120 31020 1.01 2-Pyrrolidinone C4H7NO 7.75 86.0604 85.05313 [M+H]+ 2120 8292 4.28 3,4,5-Trimethoxycinnamic acid C12H14O5 10.08 239.0893 238.08202 [M+H]+ 1110 1886 0.83 3,4-Dihydroxybenzeneacetic acid C8H8O4 6.04 169.05571 168.04843 [M+H]+ 1110 3420 -0.74 3,4-Dihydroxyphenylglycol C8H10O4 12.24 171.06263 170.05536 [M+H]+ 1100 2356 10.05 3-Aminoisobutanoic acid C4H9NO2 7.82 104.06224 103.05497 [M+H]+ 1110 560526 6.71 3-Cresotinic acid C8H8O3 9.71 153.05458 152.0473 [M+H]+ 2120 1248 2.27 3-Hydroxyanthranilic acid C7H7NO3 4 154.04743 153.04016 [M+H]+ 1110 4484 -1.3 3-Methyl-2-oxovaleric acid C6H10O3 2.91 131.07019 130.06291 [M+H]+ 2100 2318 -3.25 3a,6b,7b-Trihydroxy-5b-cholanoic acid C24H40O5 8.22 409.29264 408.28536 [M+H]+ 1110 7258 -2.55 4-Aminophenol C6H7NO 7.35 110.06005 109.05277 [M+H]+ 2120 1550 6.17 4-Ethylbenzoic acid C9H10O2 12.5 151.07537 150.06809 [M+H]+ 2120 3046 2.95 4-Hydroxyproline C5H9NO3 0.73 132.06567 131.05839 [M+H]+ 2100 1800 -0.32 5'-Methylthioadenosine C11H15N5O3S 7.23 298.09587 297.08859 [M+H]+ 2100 1266 1.59 5-Hydroxytryptophol C10H11NO2 2.76 178.08588 177.0786 [M+H]+ 2110 1150 -3.35 5-Methoxytryptophan C12H14N2O3 6.87 235.10576 234.09848 [M+H]+ 1110 2968 0.77 5-Methoxytryptophol C11H13NO2 9.33 192.10185 191.09457 [M+H]+ 2100 830 1.03 Acetaminophen C8H9NO2 7.34 152.07071 151.06352 [M+H]+, [M+Na]+ 2120 10582 1.91 Adenine C5H5N5 2.94 136.06166 135.05438 [M+H]+ 2100 1034 1.6 Adipic acid C6H10O4 1.98 129.05455 146.0579 [M-H2O+H]+, [M+H]+ 2120 4148 -3.72 ADP C10H15N5O10P2 3.31 428.03641 427.02913 [M+H]+ 2110 2336 2.05 Androstenedione C19H26O2 10.6 287.1968 286.18952 [M+H]+ 1110 9480 -1.41 Androsterone C19H30O2 17.46 291.23229 290.22502 [M+H]+ 2110 26086 4.89 Aspirin C9H8O4 10.65 181.04724 180.03996 [M+H]+ 1120 18544 2.44 Azelaic acid C9H16O4 10.02 211.09437 188.10501 [M+Na]+, [M-H2O+H]+ 2120 2410 1.75 Azythromycin C38H72N2O12 19.16 749.51278 748.50551 [M+H]+ 1100 1238 11.41 Benzaldehyde C7H6O 13.73 107.04912 106.04185 [M+H]+ 2020 13866 Cinnamaldehyde C9H8O 12.49 133.06574 132.05846 [M+H]+ 2100 930 4.95 Cortisol C21H30O5 15.18 363.21436 362.20708 [M+H]+ 1120 57574 5.95 Cyclic AMP C10H12N5O6P 5.95 330.05986 329.05259 [M+H]+ 2110 680 4.51 Cytidine C9H13N3O5 2.95 244.09309 243.08582 [M+H]+ 2110 1234 1.57 Diacetyl C4H6O2 7.42 87.04444 86.03716 [M+H]+ 2120 4060 -0.14 Dihydrothymine C5H8N2O2 12.03 129.0695 128.06223 [M+H]+ 1100 1428 8.8 Glycerol C3H8O3 2.89 93.05465 92.04737 [M+H]+ 2120 18832 1.77 Glycerophosphocholine C8H20NO6P 1.77 258.11045 257.10318 [M+H]+ 2110 2150 0.73 Glycyl-L-leucine C8H16N2O3 5.99 189.12356 188.11629 [M+H]+ 2100 1188 4.63 Guanine C5H5N5O 3.03 152.05673 151.04945 [M+H]+ 2110 4370 1.73 Guanosine C10H13N5O5 5.91 284.09885 283.09158 [M+H]+ 2110 3242 1.47 Guanosine 5'-diphosphate C10H15N5O11P2 3.04 444.03152 443.02425 [M+H]+ 2010 1574 Hydroquinone C6H6O2 2.01 111.044 110.03672 [M+H]+ 2120 1868 -4.05 Hypoxanthine C5H4N4O 5.92 137.04568 136.03841 [M+H]+ 2120 27410 4.57 Indoleacrylic acid C11H9NO2 7.24 188.07083 187.06356 [M+H]+ 2120 14526 -1.83 Inosine C10H12N4O5 5.92 269.08847 268.08119 [M+H]+ 2110 2798 1.49 Isobutyric acid C4H8O2 15.43 89.06 88.05273 [M+H]+ 2120 29906 9.05 L(-)-Nicotine pestanal C10H14N2 3.32 163.12296 162.11569 [M+H]+ 2110 1942 1.95 L-Arginine C6H14N4O2 3.32 175.11906 174.11179 [M+H]+ 2110 1844 2.38 L-Dopa C9H11NO4 3.47 198.07424 197.06696 [M+H]+ 2100 1230 2.13 L-Glutamic acid C5H9NO4 6.42 148.06038 147.0531 [M+H]+ 2120 12732 5.39 L-Glutamine C5H10N2O3 2.98 147.08449 146.07722 [M+H]+ 1100 1294 1.96 L-Histidine C6H9N3O2 9.27 156.08075 155.07347 [M+H]+ 1110 1242 8.34 L-Kynurenine C10H12N2O3 6.8 209.10196 208.09468 [M+H]+ 1120 2128 2.01 L-Methionine C5H11NO2S 3.29 150.05793 149.05065 [M+H]+ 2110 2560 1.95 L-Norleucine C6H13NO2 10.59 132.10183 131.09456 [M+H]+ 2120 11740 7.47 L-Phenylalanine C9H11NO2 6.48 166.08639 165.07912 [M+H]+ 2120 14992 1.58 L-Threonine C4H9NO3 7.82 120.05701 119.04973 [M+H]+ 1110 15840 6.82 L-Tryptophan C11H12N2O2 7.24 205.09745 204.09017 [M+H]+ 2120 3086 1.45 Maltitol C12H24O11 1.85 367.12096 344.13186 [M+Na]+, [M+K]+ 2110 161518 0.75 Metoprolol C15H25NO3 13.01 268.18852 267.18124 [M+H]+ 1110 2204 5.59 Mevalonic acid C6H12O4 3.32 171.06269 148.07347 [M+Na]+, [M-H2O+H]+, [M+H]+ 2020 195302 N,N-Dimethylformamide C3H7NO 1.86 74.06044 73.05316 [M+H]+ 2120 4234 -0.4 N-Acetyl-L-alanine C5H9NO3 2.04 132.06552 131.05817 [M+H]+, [M-H2O+H]+ 2120 19464 0.99 N-Acetyl-L-phenylalanine C11H13NO3 6.92 208.09469 207.08742 [M+H]+ 1120 2044 -0.7 N-Acetylputrescine C6H14N2O 9.97 131.11776 130.11048 [M+H]+ 2100 1006 8.73 N-Acetylserotonin C12H14N2O2 7.47 219.11097 218.10369 [M+H]+ 2110 3886 1.03 Naproxen C14H14O3 11.89 231.09933 230.09205 [M+H]+ 1110 1644 0.53 Niacinamide C6H6N2O 3.31 123.05512 122.04784 [M+H]+ 2120 10944 1.97 Norepinephrine C8H11NO3 6.14 170.08128 169.074 [M+H]+ 2100 1726 5.01 Nutriacholic acid C24H38O4 21.12 391.28186 390.27458 [M+H]+ 1110 3218 8.85 o-Tyrosine C9H11NO3 3.41 182.08152 181.07424 [M+H]+ 2100 2990 -1.25 p-Hydroxyphenylacetic acid C8H8O3 5.93 153.05928 152.052 [M+H]+ 1100 996 -1.51 p-Octopamine C8H11NO2 6.79 154.08631 153.07904 [M+H]+ 2020 1578 PC(16:0/16:0) C40H80NO8P 16.85 734.56891 733.56164 [M+H]+ 2000 1580 PC(18:1(9Z)/18:1(9Z)) C44H84NO8P 19.94 786.60247 785.59519 [M+H]+ 2000 990 Phenylacetaldehyde C8H8O 11.59 121.06475 120.05747 [M+H]+ 2120 23630 6.37 Phenylacetic acid C8H8O2 12.01 137.05974 136.05247 [M+H]+ 2120 3254 3.47 Phenylacetylglycine C10H11NO3 6.51 194.07898 193.0717 [M+H]+ 1110 10090 -1.1 Phosphocreatine C4H10N3O5P 13.73 212.03766 211.03038 [M+H]+ 1010 82948 Phosphoric acid H3O4P 2.15 98.98425 97.97698 [M+H]+ 2100 2410 1.06 Pipecolic acid C6H11NO2 6.13 130.08622 129.07894 [M+H]+ 2120 3118 4.79 Progesterone C21H30O2 9.37 315.22811 314.22083 [M+H]+ 1110 10750 -4.47 Propionic acid C3H6O2 1.96 57.03377 74.03709 [M-H2O+H]+, [M+H]+ 2120 125120 -3.24 Pyroglutamic acid C5H7NO3 6.09 130.04977 129.04249 [M+H]+ 2100 870 4.74 Pyrrole-2-carboxylic acid C5H5NO2 6.42 112.03934 111.03207 [M+H]+ 2100 864 0.41 Rhamnose C6H12O5 9.62 165.07597 164.06869 [M+H]+ 2100 1090 8.43 Riboflavin C17H20N4O6 8.92 377.15349 376.14621 [M+H]+ 1110 908 2.35 Saccharopine C11H20N2O6 13.58 277.14124 276.13396 [M+H]+ 2110 6138 12.59 Salicin C13H18O7 6.14 287.10623 286.09864 [M+H]+, [M+Na]+ 1000 2178 Serotonin C10H12N2O 11.39 177.11223 176.10495 [M+H]+ 1110 13792 9.99 Sorbitol C6H14O6 1.97 183.08648 182.07924 [M+H]+, [M-H2O+H]+ 2120 22156 0.93 Spermine C10H26N4 1.42 203.22352 202.21624 [M+H]+ 2110 5796 0.61 Sphinganine C18H39NO2 12.25 302.30551 301.29823 [M+H]+ 2120 10548 -1.14 Succinylacetone C7H10O4 10.65 159.06519 158.05791 [M+H]+ 2020 37032 Sulfolithocholic acid C24H40O6S 13.42 457.25788 456.2506 [M+H]+ 1000 1248 Taurocholic acid C26H45NO7S 12.73 516.29026 515.28299 [M+H]+ 1100 1394 3.19 Taurodeoxycholic acid C26H45NO6S 8.18 500.2951 499.28782 [M+H]+ 1100 1788 -2.56 Tetrahydrocortisone C21H32O5 7.68 365.22701 364.21973 [M+H]+ 1100 1158 -2.26 Thymidine C10H14N2O5 9.95 243.08832 242.08105 [M+H]+ 1100 1016 5.02 Tiglylglycine C7H11NO3 2.51 158.09116 157.08388 [M+H]+ 1100 1194 -3.31 Trehalose C12H22O11 9.63 343.12941 342.12214 [M+H]+ 1110 1114 8.57 Tryptophanol C10H11NO 6.99 162.09147 161.08419 [M+H]+ 2120 2420 -1.62 Uracil C4H4N2O2 3.53 113.03449 112.02722 [M+H]+ 2000 300 Urea CH4N2O 13.73 61.04 60.03272 [M+H]+ 2010 67932 Ureidosuccinic acid C5H8N2O5 14.48 177.05469 176.04741 [M+H]+ 1110 5990 13.2 Uric acid C5H4N4O3 3.31 169.03196 168.02465 [M+H]+, [M-H2O+H]+ 1010 19566 Uridine C9H12N2O6 14.48 245.07853 244.07126 [M+H]+ 2010 24674 Urocanic acid C6H6N2O2 3.31 139.05038 138.0431 [M+H]+ 2100 3066 1.97 Verapamil C27H38N2O4 8.29 455.28584 454.27856 [M+H]+ 1100 1836 -1.65 METABOLITES_END #END