SUPPLEMENTAL MATERIAL FOR “URINE METABOLITE PROFILES PREDICTIVE OF HUMAN KIDNEY ALLOGRAFT STATUS” by KARSTEN SUHRE et al.

I. SUPPLEMENTAL METHODS AND PLANNED MECHANISTIC STUDIES…………….pages 1 ‐10

II. SUPPLEMENTAL FIGURES 1, 2 and 3…………………………………………………………. pages 11‐14

III. REFERENCES…………………………………………………………………………………………………….page 15

IV. SUPPLEMENTAL TABLES 1, 2, 3, 4, 5, 6 and 7…………………………………………….. pages 16‐39

I. SUPPLEMENTAL METHODS

Collection of urine specimens. Four hundred and eighty five patients with end stage renal

disease were enrolled in the parent Clinical Trials of Transplantation‐04 (CTOT‐04) multicenter, prospective observational study.1 The following 5 clinical sites enrolled the CTOT‐04 study

participants: Northwestern University Feinberg School of Medicine, Chicago, IL (n = 145 kidney graft recipients); New York Presbyterian Hospital‐Weill Cornell Medical Center, New York, NY (n

= 122 kidney graft recipients); Hospital of University of Pennsylvania, Philadelphia, PA (n = 120 kidney graft recipients); New York Presbyterian Hospital‐Columbia University Medical Center,

New York, NY (n = 76 kidney graft recipients); and University of Wisconsin Hospital and Clinics,

Madison, WI (n = 34 kidney graft recipients).

Urine sample collection and shipment. Fifty to 100 ml of midstream urine were collected in a sterile, sealed urine collection cup without addition of any preservatives. Urine was processed within 1 hour of collection and if this was not possible, urine was refrigerated at 4° C for a maximum time of 4 hours. The urine sample was centrifuged at 2000g at room temperature for

1 of 39 30 minutes in sterile disposable tubes. After centrifugation, 6 ml of the supernatant was aspirated using a pipette without disturbing the cell pellet and transferred to four (4) 1.5 ml microcentrifuge polypropylene tubes.

The supernatants and the cell pellets were stored as specified by the NIH‐sponsored Statistical

Analysis and Clinical Coordinating Center. In brief, the supernatants and the cell pellets from

the urine specimens were first stored at the clinical site at ‐80° C and shipped in a Styrofoam container with dry ice. The samples from the ‐80° C freezer were transferred into the middle of the dry ice to avoid any thawing of the samples. The samples were then covered with more dry

ice so that the samples were completely surrounded by at least 10 cm of dry ice on all sides.

The Styrofoam container was covered with a well fitting cover and the box was sealed with duct tape. The samples were shipped to the Suthanthiran Laboratory at Weill Cornell Medical

College (WCMC, NY, NY) and stored at ‐80° C. The unthawed samples were similarly shipped from the Suthanthiran laboratory to Metabolon Inc. for non‐targeted metabolite analysis, and to the Gross laboratory at WCMC, NY, NY for measurement of 3‐sialyllactose to xanthosine ratio.

In the CTOT‐04 study, sequential urine samples were collected on post‐transplant days 3, 7, 15 and 30 and in months 2, 3, 4, 5, 6, 9 and 12 and at the time of biopsy and 2 weeks following a biopsy. A total of 4300 urine samples were prospectively collected from 485 kidney graft

recipients (patients) and urine cell pellet and cell free supernatants were prepared at the 5 clinical sites using a standard protocol summarized above.

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Urine sample selection for metabolomics. From a total of 4300 urine samples prospectively collected from the 485‐kidney allograft recipients (patients) enrolled in the parent CTOT‐04 study, 1518 urine samples were selected for metabolite analysis to include the following urine samples: (1) all biopsy‐matched urine samples, 298 samples matched to 298 kidney allograft biopsies performed in 190 patients (biopsy matched urine samples = urine samples collected from 3 days before to 1 day after the biopsy); (2) all 808 sequential samples from 112 patients that preceded a first biopsy classified using Banff classification schema2 as acute cellular rejection, antibody‐ mediated rejection, borderline changes, or other, and sequential samples that preceded No Rejection biopsies; and (3) all 412 sequential samples from 40 patients with stable graft function and who had at least 10 sequential samples collected in the first 400 days of transplantation and with sufficient RNA for urinary cell mRNA profiling. The kidney allograft recipients designated as patients with stable graft function did not undergo biopsy during the first 400 days of transplantation and met the following additional criteria: (i) average serum creatinine less than or equal to 2.0 mg per deciliter [180 micromol per liter] at 6, 9 and 12 months following transplantation, (ii) no treatment for acute rejection, and (iii) no evidence of

cytomegalovirus (CMV) or polyomavirus type BK (BKV) infection.

The number of patients in the three categories listed under the metabolomics (Figure 1) exceeds 241 unique patients because several patients had multiple urine samples and contributed urine samples to more than one category, that is the same patient contributing biopsy matched urine sample as well as sequential urine samples and being counted in both categories.

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Two of the 1,518 samples were excluded from all analyses: one biopsy‐matched urine sample

(sample ID: 4202) from a patient (patient ID: 10137) with a No Rejection biopsy result was excluded due to failed osmolality measurements and only 80 metabolites being measured in that sample; and one of the sequential, non‐biopsy associated samples from a patient with other biopsy findings (urine sample ID:0988; patient ID: 10014) did not contain sufficient cell free supernatant for non targeted metabolite analysis. After exclusion of these two samples, high‐quality metabolite data from 1516 samples collected from 241 kidney patients were available for data analysis

Among the 298 biopsy matched urine samples, all 50 ACR biopsy matched urine samples from

36 patients and all 198 No Rejection biopsy urine samples from 149 patients were included in the analysis to determine whether metabolite profiles are diagnostic of acute cellular rejection.

Urine samples matched to biopsies classified as borderline changes (27 samples from 25 patients), AMR (13 samples from 11 patients) and other findings (9 samples from 8 patients) were all also analyzed but excluded from the analyses to generate the diagnostic signature distinguishing ACR biopsies from No Rejection biopsies. The sequential samples preceding borderline changes, AMR and other findings were also not included in the analysis to determine whether the signature distinguishes patients who developed ACR from those whose biopsies did not show rejection changes (No Rejection biopsies).

4 of 39 A total of 2782 urine samples collected during the parent CTOT‐04 study were excluded from

non‐targeted metabolite analysis because: (1) the urine samples were not matched to biopsy specimens or were collected following a biopsy (1074 specimens from 187 patients), (2) had unstable allograft function and no biopsy or were lost to follow up (272 urine samples from 63 patients) or (3) had stable graft function but had less than 10 sequential samples collected in

the first 400 days of transplantation; urine samples collected after 400 days and with insufficient RNA for mRNA profiling are also included in this group of 1436 specimens from 180 patients. The number of patients in the three categories excluded from metabolomics exceeds

243 unique patients because several patients had multiple urine samples and contributed urine samples to more than one category and thereby counted in each category.

Metabolomics data acquisition. The extracted samples were split into equal parts for analysis on the Gas Chromatography / Mass Spectrometry and Liquid Chromatography/ Tandem Mass

Spectrometry platforms. Also included were several technical replicate samples created from a homogeneous pool containing a small amount of all study samples (“Client Matrix”).

Instrument variability was determined by calculating the median relative standard deviation

(RSD) for the internal standards that were added to each sample prior to injection into the mass spectrometers. Overall process variability was determined by calculating the median RSD for all endogenous metabolites (i.e., non‐instrument standards) present in 100% of the Client Matrix samples, which are technical replicates of pooled client samples. Metabolites of known structural identity (named biochemicals) as well as metabolites of unknown structural identity

(unnamed biochemicals) detected in the samples are reported. Information related to

5 of 39 biochemical pathways, chemical properties, and public databases are included in Supplemental

Table 1.

Sample handling. Samples were accessioned into the Metabolon Laboratory Information

Management System (LIMS) and were assigned a unique identifier by the LIMS, which was associated with the original source identifier only. This identifier was used to track all sample handling, tasks, results, etc. The samples (and all derived aliquots) were bar‐coded and tracked by the LIMS system. All portions of any sample were automatically assigned their own unique identifiers by the LIMS when a new task was created; the relationships of these sample portions were also tracked. All samples were maintained at ‐80C until processed. Sample preparation was carried out using an automated system (MicroLab STAR, Hamilton Robotics). Recovery

standards were added prior to the first step in the extraction process for QC purposes. Sample preparation was conducted using a proprietary series of organic and aqueous extractions to remove the protein fraction while allowing maximum recovery of small molecules. The resulting extract was divided into two fractions; one for analysis by LC and one for analysis by GC.

Samples were placed briefly on a TurboVap (Zymark) to remove the organic solvent. Each

sample was then frozen, dried under vacuum, and prepared for the appropriate instrument,

either LC/MS or GC/MS. For QA/QC purposes, a number of additional samples were included with each day’s analysis. Furthermore, a selection of QC metabolites was added to every sample, including those under test. These metabolites were carefully chosen so as not to interfere with the measurement of the endogenous metabolites.

6 of 39 Liquid Chromatography/Mass Spectrometry (LC/MS) and Liquid Chromatography / Tandem

Mass Spectrometry (LC/MS2) for Metabolite Profiling. The LC/MS portion of the platform was based on a Waters ACQUITY UPLC and a Thermo‐Finnigan LTQ mass spectrometer, which consisted of an electrospray ionization (ESI) source and linear ion‐trap (LIT) mass analyzer. The dried sample extract was split into two aliquots, and then reconstituted in acidic or basic LC‐ compatible solvents, each of which contained 11 or more injection standards at fixed concentrations. One aliquot was analyzed using acidic positive ion optimized conditions and the

other using basic negative ion optimized conditions in two independent injections using

separate dedicated columns (Waters UPLC BEH C18‐2.1x100 mm, 1.7 µm). Extracts

reconstituted in acidic conditions were gradient eluted using water and methanol both

containing 0.1% formic acid, while the basic extracts, which also used water/methanol, contained 6.5 mM ammonium bicarbonate. The MS analysis alternated between MS and data‐ dependent MS2 scans, using dynamic exclusion, and the scan range was from 80‐1000 m/z. For ions with counts greater than 2 million, an accurate mass measurement could be performed. Accurate mass measurements were made for the parent ion as well as fragments. The typical mass error was less than 5 ppm. Ions with less than two million counts require a greater amount of effort to characterize. Fragmentation spectra (MS/MS) were typically generated in a data dependent manner, but if necessary, targeted MS/MS was employed, such as in the case of lower level signals.

Gas Chromatography/Mass Spectrometry (GC/MS) for Metabolite Profiling. The samples destined for GC/MS analysis were re‐dried under vacuum desiccation for a minimum of 24

7 of 39 hours prior to being derivatized under dried nitrogen using bistrimethyl‐silyl‐triflouroacetamide

(BSTFA). The GC column was 5% diphenyl / 95% dimethyl polysiloxane fused silica column (20

m x 0.18 mm ID; 0.18 um film thickness) with helium as the carrier gas and a temperature ramp from 60° to 340°C over a 17.5‐min period. Samples were analyzed on a Thermo‐Finnigan Trace

DSQ fast‐scanning single‐quadruple mass spectrometer using electron impact ionization and

operated at unit mass resolving power. The scan range was 50‐750 m/z. The instrument was tuned and calibrated for mass resolution and mass accuracy on a daily basis.

Metabolite identification. The data extraction of the raw mass spec data files yielded information that was loaded into a relational database. Once in the database the information was examined and appropriate QC limits were imposed. Peaks were identified using

Metabolon’s proprietary peak integration software, and component parts were stored in a separate, specifically designed complex data structure. Metabolites were identified by

comparison to library entries of purified standards or recurrent unknown entities. More than

3,500 commercially available purified standard compounds have been acquired and registered

into the LIMS for distribution to both the LC and GC platforms for determination of their analytical characteristics. The combination of chromatographic properties and mass spectra gave an indication of a match to the specific compound or an isobaric entity. Additional entities could be identified by virtue of their recurrent nature (both chromatographic and mass spectral). These compounds have the potential to be identified by future acquisition of a matching purified standard or by classical structural analysis.3 A variety of curation procedures were carried out to ensure that a high quality data set was made available for statistical analysis

8 of 39 and data interpretation. The QC and curation processes were designed to ensure accurate and consistent identification of true chemical entities, and to remove those representing system artifacts, mis‐assignments, and background noise. Metabolon data analysts use proprietary visualization software and interpretation software to confirm the consistency of peak identification among the various samples. Library matches for each compound were checked in

each sample and corrected if necessary.

Metabolomics data. In total, 749 different metabolites from 65 metabolic pathways, including

368 metabolites of unknown identity, were identified on at least one of the three mass spectrometry based metabolomics platforms (for details see Supplemental Table 1). 358 metabolites were reported from the LC/MS platform running in negative ionization mode, 245 using LC/MS in positive mode, and 146 using GC/MS. The median number of different metabolites detected in any single sample was 616. The median technical error as determined from technical replicates on pooled sample material (s.d./mean) was 10.2%, while the median experimental variance computed from data on all patient samples was 113.9%, showing overall excellent signal‐to‐noise performance.

Stability of metabolites in urine. The stability of metabolites in stored samples is an important issue and we have not analyzed urine supernatants stored for different duration. The data we provide in this report however suggests that urine supernatants stored over several years yield

robust metabolite signatures diagnostic of acute cellular rejection. The first urine specimen included in the 1516 samples analyzed in our study for metabolites was collected in 2006 and

9 of 39 the last urine specimen included in our study was collected in 2009. Non‐targeted metabolite analysis of the stored urine supernatants was performed in year 2012 at Metabolon Inc. and showed that the metabolite profiles including the ratio of 3‐sialyllactose to xanthosine are diagnostic of acute cellular rejection. Rapid Fire‐QTOF MS/MS assay for the measurement of 3‐ sialyllactose to xanthosine ratio was developed in the Gross laboratory at WCMC, NY, NY

(Supplemental Figure 1) and the assays performed in 2014 demonstrated that the ratio of of 3‐ sialyllactose to xanthosine is diagnostic of acute cellular rejection (Supplemental Figure 2).

Importantly, the observed correlation between the 3‐sialyllactose to xanthosine ratio calculated from the Metabolon data and the 3‐sialyllactose to xanthosine ratio measured using RapidFire‐

QTOF assay was 0.65 (Pearson R), and the Bland‐Altman method for comparison showed that only 12 samples (6%) were beyond the 95% limit of agreement (Supplemental Figure 3).

Mechanistic basis for the metabolite signatures. Our urine metabolomics study discovered

that the ratio of 3‐sialyllactose to xanthosine and the ratio of quinolinate to X‐16397 were best

associated with ACR biopsy diagnosis. We plan to perform the following studies to elucidate the

basis for the relationship between the signatures and biopsy diagnosis: (i) examine the effect of

mycophenolic acid, tacrolimus and corticosteroids on the levels of 3‐sialyllactose, xanthosine

and quinolinate in cell free supernatants of human peripheral blood cells incubated with or without anti‐CD3 mABs, and in cell free supernatants of human renal tubular cells incubated with or without inflammatory cytokines; (ii) investigate the changes in sialyllactose levels in cells grown under oxidative stress; and (iii) structural resolution to identify the currently unknown metabolite X‐16397.

10 of 39 II. SUPPLEMENTAL FIGURES

Supplemental Figure 1.

Supplemental Figure 1. Development of RapidFire‐QTOF MS/MS assay for simultaneous quantification of 3‐sialyllactose, xanthosine, neopterin and creatinine in a urine matrix. Results show linearity for detection for spiked in pure molecule standards and absolute quantification of endogenous levels of the cognate urinary metabolite as the Y‐axis intercept.

11 of 39 Urine samples were diluted 10‐fold in 70:30 acetonitrile/H2O containing 0.2% ammonium hydroxide. Mean and median osmolality of all CTOT urine samples analyzed were 483.5 and 453 mOsm/kg, respectively. Diluted urine samples were briefly vortexed and centrifuged at 16000 x g in a microfuge to pellet precipitated proteins and debris. Sample supernatants were transferred to 96 well plates and diluted 1:1 with H2O for subsequent quantification of 3‐ sialyllactose: xanthosine ratios using robotic solid‐phase extraction and tandem MS detection. Analyte quantification was performed using a RapidFire 365 automated sampling module (Agilent Technologies) in combination with an Agilent 6550 iFunnel Quadrupole Time‐of‐Flight (QTOF) mass spectrometer. Desalting of samples and concentration of urinary 3‐sialyllactose and xanthosine was achieved using a graphite‐carbon (type D) mini‐SPE cartridge (Agilent Technologies). Analysis of each sample was completed in a total of 13.6 s, including time allotments as follows: sample aspiration/uptake, 0.6 s; sample loading and cartridge washing, 3 s; sample elution, 7 s; and cartridge re‐equilibration, 3 s. An aqueous mobile phase containing 5 mM ammonium acetate (NH4Ac) was used during the sample loading and washing phase, while

an organic mobile phase containing 5 mM NH4Ac in 80:20 acetonitrile/ H2O was used during the elution and cartridge re‐equilibration phases. The mobile phase flow rates for loading/washing, elution, and re‐equilibration phases were 1.5 ml/min; 1.0 ml/min; and 1.25 ml/min, respectively. The total injected sample volume was 10 µl. The QTOF was calibrated in 2 GHz (extended dynamic range) mode. The product ion mass spectra of sialyllactose and xanthosine were acquired in negative ion mode at 8 spectra/sec, sampled over a mass/charge range of 40– 700 Daltons. The source parameters were: capillary voltage, 3500 V; nozzle voltage, 1000V; fragmentor voltage, 140 V; skimmer, 65; and OctapoleRFPeak, 750. The nitrogen drying gas and sheath gas temperatures were 200°C and 350˚C, respectively. The nebulizer pressure was maintained at 35 psi, delivered at a flow rate of 14 l/min. Sheath gas flow was 11 l/min. The collision energy for product ion generation from sialyllactose (C11H17NO8, m/z, 290.0881) and xanthosine (C5H4N4O2, m/z, 151.0262) was 30 ev and 20 ev, respectively. Product ion data was analyzed using Agilent MassHunter Quantitative Analysis software B6.0. build 6.0. 388.0. The ratio of sialyllactose to xanthosine was calculated from the peak area of the two product ions.

12 of 39 Supplemental Figure 2.

P=4.04e-8 6

4

2

0 ACR Normal

Sialyllactose:Xanthosine

Supplemental Figure 2. The ratio of 3‐sialyllactose to Xanthosine in the biopsy matched urine samples, measured using the RapidFire‐QTOF MS/MS assay, is diagnostic of acute cellular rejection in the human kidney allograft. Pristine (not previously thawed) aliquots of urine supernatants were used for measurement of 3SL/X ratio in the RapidFire‐QTOF MS/MS assay. Each supernatant was assayed in 2 or 3 replicate wells and the mean of the replicates was calculated and used in the data analysis. The 3SL/X ratio in 206 CTOT‐04 biopsy‐matched urine samples (43 ACR biopsy‐matched samples and 163 No Rejection biopsy‐matched samples) selected for urine metabolomics study was measured. The P value for the ability of the ratio of 3SL/X to discriminate ACR biopsies from No Rejection biopsies was P=4.0x10‐8 (Mann –Whitney test) with the targeted RapidFire assay. The results based on the Rapid Fire assay are in accord with those based on the 3SL/X ratio based on the non‐targeted metabolite data generated by Metabolon, Inc.

13 of 39

Supplemental Figure 3.

Supplemental Figure 3. Bland‐Altman plot showing the relationship between the 3SL/X ratio calculated from the non‐targeted Metabolite analysis performed by Metabolon Inc. and the 3SL/X ratio measured using targeted RapidFire assay in the Gross laboratory. The observed correlation between the 3SL/X ratio calculated from the non‐targeted Metabolite analysis performed by Metabolon Inc. and the 3SL/X ratio measured using targeted RapidFire assay in the Gross laboratory was 0.65 (Pearson R) and the Bland‐Altman method for comparison4 showed that only 12 samples (6%) were beyond the 95% limit of agreement.

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III. REFERENCES

1. Suthanthiran, M, Schwartz, JE, Ding, R, Abecassis, M, Dadhania, D, Samstein, B, Knechtle, SJ,

Friedewald, J, Becker, YT, Sharma, VK, Williams, NM, Chang, CS, Hoang, C, Muthukumar,

T, August, P, Keslar, KS, Fairchild, RL, Hricik, DE, Heeger, PS, Han, L, Liu, J, Riggs, M, Ikle,

DN, Bridges, ND, Shaked, A: Urinary‐cell mRNA profile and acute cellular rejection in

kidney allografts. N Engl J Med 369: 20‐31, 2013

2. Racusen, LC, Colvin, RB, Solez, K, Mihatsch, MJ, Halloran, PF, Campbell, PM, Cecka, MJ,

Cosyns, JP, Demetris, AJ, Fishbein, MC, Fogo, A, Furness, P, Gibson, IW, Glotz, D, Hayry,

P, Hunsickern, L, Kashgarian, M, Kerman, R, Magil, AJ, Montgomery, R, Morozumi, K,

Nickeleit, V, Randhawa, P, Regele, H, Seron, D, Seshan, S, Sund, S, Trpkov, K: Antibody‐

mediated rejection criteria ‐ an addition to the Banff 97 classification of renal allograft

rejection. Am J Transplant 3: 708‐714, 2003

3. Dehaven, CD, Evans, AM, Dai, H, Lawton, KA: Organization of GC/MS and LC/MS

metabolomics data into chemical libraries. J Cheminform 2: 9, 2010

4. Bland, JM, Altman, DG: Statistical methods for assessing agreement between two methods of

clinical measurement. Lancet 1: 307‐310, 1986

15 of 39 IV. SUPPLEMENTAL TABLES

The reported association data in Supplemental Tables 3‐7 is limited to p<0.05 or p‐gain>750 Bonferroni significance level for metabolites p<0.05/(749)=6.68E‐5 Bonferroni significance level for ratios p<0.05/(749*748/2)=1.76E‐7 Bonferroni significance for p‐gain p‐gain > 10*749 = 7,490

Supplemental Table 1 Metabolite summary statistics and descriptors Supplemental Table 2 Characteristics of CTOT‐04 Kidney Transplant Recipients Included in the Analysis

Supplemental Table 3 Linear regression [Mi = f(ACR)] using all samples with matching biopsy

Supplemental Table 4 Logistic regression [ACR = f(Mi)] using all samples with matching biopsy Supplemental Table 5 Linear regression [Mi = f(ACR)] using all samples with matching biopsy and available mRNA signature

Supplemental Table 6 Logistic regression [ACR = f(Mi)] using all samples with matching biopsy and available mRNA signature

Supplemental Table 7 Logistic regression [ACR = f(Mi, RNA_signature)] using all samples with matching biopsy and available mRNA signature

Supplemental Table 1 collumn header description IND index to sort by pathway COMP_ID Metabolon compound identifier BIOCHEMICAL metabolite name SUPER_PATHWAY metabolite super pathway SUB_PATHWAY metabolite pathway PLATFORM platform used to detect this metabolite (LC/MS Pos, LC/MS Neg, GC/MS) RI retention index MASS ion mass PUBCHEM pubchem identifier CAS CAS identifier KEGG KEGG identifier HMDB_ID HMDB identifier # samples number of urine samples in which this metabolite was detected RSD relative technical variance (%, based on Metabolon RAW data) EXP relative experimental variance (%, based on Metabolon RAW data) EXP/RSD ratio between experimental and technical variance

Supplemental Tables 3 & 5 collumn header description numerator Metabolon compound identifier in the numerator denominator Metabolon compound identifier in the denominator ('one' indicates 'no ratio') ncases number of data points used in the statistics beta Differences in mean levels of metabolites between ACR and No Rejection groups p‐value p‐value of the association with ACR p‐gain p‐gain of the association with ACR (when using ratios)

Supplemental Tables 4 & 6 collumn header description numerator Metabolon compound identifier in the numerator denominator Metabolon compound identifier in the denominator ('one' indicates 'no ratio') ncases number of data points used in the statistics OR odds ratio of the association with ACR, per 1 sd change in predictor p‐value p‐value of the association with ACR p‐gain p‐gain of the association with ACR (when using ratios)

Supplemental Table 7 collumn header description numerator Metabolon compound identifier in the numerator denominator Metabolon compound identifier in the denominator ('one' indicates 'no ratio') ncases number of data points used in the statistics OR(metabolite) odds ratio of the association with ACR, per 1 sd change in predictor p‐value(metabolite) p‐value for metabolite p‐gain(metabolite) p‐gain of the metabolite (when using ratios) OR(mRNA signature) odds ratio of the mRNA signature p‐value(mRNA signature) p‐value of the mRNA signature

16 of 39 Supplemental Table 1 Metabolite summary statistics and descriptors

IND COMP_ID BIOCHEMICAL SUPER_PATHWAY SUB_PATHWAY PLATFORM RI MASS PUBCHEM CAS KEGG HMDB_ID # samples RSD EXP EXP/RSD 1 11777 glycine Amino acid Glycine, serine and threonine metabolism GC/MS 1166 101.9 5,257,127,750 56‐40‐6; C00037 HMDB00123 1517 15.6 103.6 6.6 2 1516 sarcosine (N‐Methylglycine) Amino acid Glycine, serine and threonine metabolism GC/MS 1182.9 116 10,887,311,726 107‐97‐1; C00213 HMDB00271 1513 13.7 79 5.8 3 5086 dimethylglycine Amino acid Glycine, serine and threonine metabolism GC/MS 1030 57.9 6,971,056,673 1118‐68‐9; C01026 HMDB00092 1484 11.7 91 7.8 4 27710 N‐acetylglycine Amino acid Glycine, serine and threonine metabolism GC/MS 1396.4 143.9 10972 543‐24‐8; HMDB00532 1298 24.5 192.9 7.9 5 15686 beta‐hydroxypyruvate Amino acid Glycine, serine and threonine metabolism GC/MS 1425.1 221 964 1113‐60‐6;3369‐79‐7; C00168 HMDB01352 1203 43.2 95.8 2.2 6 1648 serine Amino acid Glycine, serine and threonine metabolism GC/MS 1389.1 204 59,516,857,581 56‐45‐1; C00065 HMDB00187 1516 14.9 99.4 6.7 7 37076 N‐acetylserine Amino acid Glycine, serine and threonine metabolism GC/MS 1526 218 65249 97‐14‐3; HMDB02931 1211 27.7 105.2 3.8 8 23642 homoserine Amino acid Glycine, serine and threonine metabolism GC/MS 1471 218.1 126,476,971,022 672‐15‐1; C00263 HMDB00719 1257 22.2 121.3 5.5 9 1284 threonine Amino acid Glycine, serine and threonine metabolism GC/MS 1412.3 218.1 69,710,196,288 72‐19‐5; C00188 HMDB00167 1516 11.7 174.8 14.9 10 15142 allo‐threonine Amino acid Glycine, serine and threonine metabolism GC/MS 1417.8 219 992,896,995,276 28954‐12‐3; C05519 HMDB04041 1507 19 62.6 3.3 11 15996 aspartate Amino acid Alanine and aspartate metabolism GC/MS 1529.7 232 5960 56‐84‐8; C00049 HMDB00191 1403 92.8 356.2 3.8 12 11398 asparagine Amino acid Alanine and aspartate metabolism GC/MS 1651.2 231 62,676,992,089 70‐47‐3; C00152 HMDB00168 1421 19.9 166.4 8.4 13 33942 N‐acetylasparagine Amino acid Alanine and aspartate metabolism LC/MS Pos 974 175.1 99715 4033‐40‐3; HMDB06028 1496 19.5 65.2 3.3 14 55 beta‐alanine Amino acid Alanine and aspartate metabolism GC/MS 1451.8 174 2,394,755,801 56‐41‐7;107‐95‐9; C00099 HMDB00056 1455 27.9 241.2 8.6 15 3155 3‐ureidopropionate Amino acid Alanine and aspartate metabolism LC/MS Pos 1062 133.1 111 462‐88‐4; C02642 HMDB00026 1509 22.4 72.7 3.2 16 37432 N‐acetyl‐beta‐alanine Amino acid Alanine and aspartate metabolism LC/MS Pos 1460 132.1 76406 3025‐95‐4; C01073 1500 16.3 77.4 4.7 17 1126 alanine Amino acid Alanine and aspartate metabolism GC/MS 1147.6 115.9 59,507,311,724 56‐41‐7; C00041 HMDB00161 1517 10.4 116.7 11.2 18 22185 N‐acetylaspartate (NAA) Amino acid Alanine and aspartate metabolism LC/MS Pos 1222 176.1 65065 997‐55‐7;997‐55‐7 ; C01042 HMDB00812 1158 22.2 71.7 3.2 19 57 glutamate Amino acid Glutamate metabolism GC/MS 1611.9 246 611 56‐86‐0; C00025 HMDB03339 1513 19.4 273.7 14.1 20 53 glutamine Amino acid Glutamate metabolism LC/MS Pos 684 147.2 5961 56‐85‐9; C00064 HMDB00641 1516 5.1 123 24.1 21 32672 pyroglutamine* Amino acid Glutamate metabolism LC/MS Pos 764 129.2 134508 2353‐44‐8; 1495 26.9 94.1 3.5 22 33943 N‐acetylglutamine Amino acid Glutamate metabolism LC/MS Neg 783 187.1 182230 2490‐97‐3; C02716 HMDB06029 1500 6.9 76.8 11.1 23 59 histidine Amino acid Histidine metabolism LC/MS Neg 757 154.1 7,733,651,426 5934‐29‐2; C00135 HMDB00177 1515 4.6 70.1 15.2 24 33946 N‐acetylhistidine Amino acid Histidine metabolism LC/MS Neg 913 196.2 75619 39145‐52‐3; C02997 1483 8.1 58.4 7.2 25 607 trans‐urocanate Amino acid Histidine metabolism LC/MS Pos 1119 139.1 736715 104‐98‐3; C00785 HMDB00301 1512 17.3 82.1 4.7 26 15677 3‐methylhistidine Amino acid Histidine metabolism LC/MS Neg 911 168.1 649,696,971,061 368‐16‐1; C01152 HMDB00479 1513 4.4 102.8 23.4 27 30460 1‐methylhistidine Amino acid Histidine metabolism GC/MS 1757.7 196 921,057,020,397 332‐80‐9; C01152 HMDB00001 1334 43.5 89.8 2.1 28 40473 hydantoin‐5‐propionic acid Amino acid Histidine metabolism LC/MS Pos 1506 173.2 782 5624‐26‐0; C05565 HMDB01212 1412 18.4 81.3 4.4 29 15716 imidazole lactate Amino acid Histidine metabolism LC/MS Pos 778 157.1 440129 14403‐45‐3; C05568 HMDB02320 1514 18.7 72.5 3.9 30 32350 1‐methylimidazoleacetate Amino acid Histidine metabolism GC/MS 1543 168.1 75810 2625‐49‐2 ; C05828 HMDB02820 1471 12.9 125.8 9.8 31 40730 imidazole propionate Amino acid Histidine metabolism LC/MS Pos 1011.7 141.1 70630 1074‐59‐5; HMDB02271 1514 14.4 124.4 8.6 32 32349 4‐imidazoleacetate Amino acid Histidine metabolism LC/MS Pos 747 127.1 96215 645‐65‐8 ; C02835 HMDB02024 1343 21.5 66 3.1 33 396 glutarate (pentanedioate) Amino acid Lysine metabolism GC/MS 1433.6 261 4418048 110‐94‐1; C00489 HMDB00661 1509 17.6 106.1 6.0 34 36863 3‐hydroxyglutarate Amino acid Lysine metabolism GC/MS 1577 259.1 181976 638‐18‐6; HMDB00428 1497 12.2 81.7 6.7 35 38667 3‐methylglutaconate Amino acid Lysine metabolism GC/MS 1462.7 198 1551553 5746‐90‐7; HMDB00522 1506 14 335.5 24.0 36 1301 lysine Amino acid Lysine metabolism LC/MS Pos 603 147.2 5962 56‐87‐1; C00047 HMDB00182 1515 14.1 120.3 8.5 37 6146 2‐aminoadipate Amino acid Lysine metabolism LC/MS Pos 794 162.1 469 542‐32‐5;1118‐90‐7; C00956 HMDB00510 1304 17.2 61.5 3.6 38 1444 pipecolate Amino acid Lysine metabolism LC/MS Pos 1120 130.1 849 4043‐87‐2; C00408 HMDB00070 1447 23.1 252.5 10.9 39 1498 N‐6‐trimethyllysine Amino acid Lysine metabolism GC/MS 1583 318.2 159659 23284‐33‐5; C03793 HMDB01325 1449 28.6 84.6 3.0 40 36751 N2‐acetyllysine Amino acid Lysine metabolism LC/MS Neg 858 187.1 699,269,792,907 1946‐82‐3; C12989 HMDB00446 1469 13 57.5 4.4 41 36752 N6‐acetyllysine Amino acid Lysine metabolism LC/MS Neg 1056 187.1 699,197,892,832 692‐04‐6; C02727 HMDB00206 1512 15.5 50.8 3.3 42 35439 glutarylcarnitine (C5) Amino acid Lysine metabolism LC/MS Pos 1565 276.1 102636‐82‐8; HMDB13130 1513 7.1 57.7 8.1 43 22130 phenyllactate (PLA) Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 2237 165.1 3848 828‐01‐3; C05607 HMDB00779 1036 12.2 152 12.5 44 64 phenylalanine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 2056 166.1 69,256,656,140 63‐91‐2; C00079 HMDB00159 1516 10.5 98.9 9.4 45 36103 p‐cresol sulfate Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 2896 187.1 4615422 3233‐57‐7; C01468 HMDB11635 1516 8 138.8 17.4 46 36845 o‐cresol sulfate Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 2821 187 11615528 836 11 159.8 14.5 47 18280 gentisate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1734 355.1 3469 490‐79‐9; C00628 HMDB00152 1487 12 159.4 13.3 48 1299 tyrosine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 1516 182.1 60,576,942,100 60‐18‐4; C00082 HMDB00158 1516 13.6 96.3 7.1 49 1431 3‐(4‐hydroxyphenyl)lactate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1823 179 9378 6482‐98‐0; C03672 HMDB00755 1486 13.8 117.3 8.5 50 1567 vanillylmandelate (VMA) Amino acid Phenylalanine & tyrosine metabolism GC/MS 1806.8 297.1 1245 55‐10‐7; C05584 HMDB00291 1517 10.5 97.2 9.3 51 1603 tyramine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 1503 138.1 5610 60‐19‐5; C00483 HMDB00306 1252 15.2 154.7 10.2 52 1669 4‐hydroxyphenylpyruvate Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 1716 179.1 979 156‐39‐8; C01179 HMDB00707 1426 20.3 130.3 6.4 53 1101 homovanillate (HVA) Amino acid Phenylalanine & tyrosine metabolism GC/MS 1731.3 209 6950168 306‐08‐1; C05582 HMDB00118 1515 10.5 66.1 6.3 54 541 4‐hydroxyphenylacetate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1630.6 179 4693933 156‐38‐7; C00642 HMDB00020 1516 10 149.6 15.0 55 1413 3‐hydroxyphenylacetate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1605.5 164 6950813 621‐37‐4; C05593 HMDB00440 1507 10.1 134.9 13.4 56 18296 3,4‐dihydroxyphenylacetate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1773.5 384.1 547 102‐32‐9; C01161 HMDB01336 1502 13.6 93.8 6.9 57 12017 3‐methoxytyrosine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 1765 212.1 9307 300‐48‐1; HMDB01434 1265 17 58.9 3.5 58 33950 N‐acetylphenylalanine Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 2590 206.2 74839 2018‐61‐3; C03519 HMDB00512 1486 8.2 117.7 14.4 59 32390 N‐acetyltyrosine Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 1677 222.2 68310 537‐55‐3 ; HMDB00866 1503 13.7 173.1 12.6 60 33945 phenylacetylglycine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 3125 194.1 68144 500‐98‐1; C05598 HMDB00821 1186 10.9 80.2 7.4 61 35126 phenylacetylglutamine Amino acid Phenylalanine & tyrosine metabolism LC/MS Pos 2868 265.2 306137 28047‐15‐6; C04148 HMDB06344 1516 2.5 37.1 14.8 62 35632 2‐(4‐hydroxyphenyl)propionate Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 1880 165.1 102526 938‐96‐5; C03080 153 11.4 87.7 7.7 63 35635 3‐(3‐hydroxyphenyl)propionate Amino acid Phenylalanine & tyrosine metabolism GC/MS 1701 310 91 621‐54‐5; C11457 HMDB00375 1088 21.4 206.6 9.7 64 32553 phenol sulfate Amino acid Phenylalanine & tyrosine metabolism LC/MS Neg 2150 173.1 74426 937‐34‐8; C02180 1515 10.7 140.6 13.1 65 42040 5‐hydroxymethyl‐2‐furoic acid Amino acid Phenylalanine & tyrosine metabolism GC/MS 1559.1 271.1 80642 6338‐41‐6; HMDB02432 1475 10.6 317.9 30.0 66 15679 xanthurenate Amino acid Tryptophan metabolism GC/MS 2000.9 406.1 5699 59‐007; C02470 HMDB00881 1511 14.4 124.6 8.7 67 1417 kynurenate Amino acid Tryptophan metabolism LC/MS Neg 2243 188.1 3845 492‐27‐3; C01717 HMDB00715 1515 5.3 79.4 15.0 68 15140 kynurenine Amino acid Tryptophan metabolism LC/MS Pos 1902 209.1 1,611,666,971,029 2922‐83‐0; C00328 HMDB00684 1507 12.4 107 8.6 69 54 tryptophan Amino acid Tryptophan metabolism LC/MS Pos 2445 205.1 69,235,166,305 73‐22‐3; C00078 HMDB00929 1516 8.8 87.4 9.9 70 18349 indolelactate Amino acid Tryptophan metabolism GC/MS 1964.9 202 92904 832‐97‐3; C02043 HMDB00671 1516 12.8 91.1 7.1 71 27513 indoleacetate Amino acid Tryptophan metabolism LC/MS Pos 3760 176.1 802 6505‐45‐9; C00954 HMDB00197 1493 10.1 114.8 11.4 72 37097 tryptophan betaine Amino acid Tryptophan metabolism LC/MS Pos 2464 247.1 442106 20671‐76‐5; C09213 1139 8.3 111 13.4 73 22110 3‐hydroxykynurenine Amino acid Tryptophan metabolism LC/MS Pos 1453 225.1 89 2147‐61‐7; C02794 HMDB00732 1009 18.4 261.9 14.2 74 32353 3‐hydroxyanthranilate Amino acid Tryptophan metabolism LC/MS Pos 2280 154.1 86 548‐93‐6; C00632 HMDB01476 1418 13 111.4 8.6 75 33959 N‐acetyltryptophan Amino acid Tryptophan metabolism LC/MS Pos 3553 247.2 700653 1218‐34‐4; C03137 1273 11.9 645.2 54.2 76 32675 C‐glycosyltryptophan* Amino acid Tryptophan metabolism LC/MS Pos 1912 367.1 1516 6.6 51.2 7.8 77 437 5‐hydroxyindoleacetate Amino acid Tryptophan metabolism GC/MS 1983.2 290 1826 54‐16‐0; C05635 HMDB00763 1516 11 86.6 7.9 78 27672 3‐indoxyl sulfate Amino acid Tryptophan metabolism LC/MS Neg 2258 212 10258 2642‐37‐7; HMDB00682 1513 12.3 86.7 7.0 79 42087 indoleacetylglutamine Amino acid Tryptophan metabolism LC/MS Neg 2530 302.2 1512 4.7 137.1 29.2 80 15676 3‐methyl‐2‐oxovalerate Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 2106 129.2 47 51829‐07‐3; C00671 HMDB03736 1261 15.2 94.6 6.2 81 12129 beta‐hydroxyisovalerate Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 1043 117.1 69362 625‐08‐1; HMDB00754 1479 7.8 99.7 12.8 82 1125 isoleucine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 1614 132.1 791 73‐32‐5; C00407 HMDB00172 1516 16.1 172.8 10.7 83 60 leucine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 1674 132.2 70,457,986,106 61‐90‐5; C00123 HMDB00687 1517 14.4 157.9 11.0 84 1557 3‐methylglutarate Amino acid Valine, leucine and isoleucine metabolism GC/MS 1450.9 275.1 12284 626‐51‐7; HMDB00752 1501 15.9 207.2 13.0 85 1587 N‐acetylleucine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 3444 174.1 70912 1188‐21‐2; C02710 HMDB11756 1251 26.3 46.5 1.8 86 1591 N‐acetylvaline Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 1697 158.2 66789 96‐81‐1; HMDB11757 1495 9.1 54 5.9 87 33967 N‐acetylisoleucine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 3370 174.1 2802421 3077‐46‐1; 1365 17.8 59.6 3.3 88 1598 tigloylglycine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2523 158.1 6441567 35842‐45‐6; HMDB00959 1512 11.7 76.3 6.5 89 1649 valine Amino acid Valine, leucine and isoleucine metabolism GC/MS 1256.3 143.9 69,710,186,287 72‐18‐4; C00183 HMDB00883 1517 19.2 281.6 14.7 90 1549 3‐hydroxyisobutyrate Amino acid Valine, leucine and isoleucine metabolism GC/MS 1205 177.11 11966314 2068‐83‐9; C06001 HMDB00336 1515 11.3 90.3 8.0 91 22116 4‐methyl‐2‐oxopentanoate Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 2200 129.2 70 816‐66‐0; C00233 HMDB00695 942 11.9 88.2 7.4 92 32397 3‐hydroxy‐2‐ethylpropionate Amino acid Valine, leucine and isoleucine metabolism GC/MS 1269 247.1 188979 4374‐62‐3; HMDB00396 1478 13 119.2 9.2 93 33937 alpha‐hydroxyisovalerate Amino acid Valine, leucine and isoleucine metabolism GC/MS 1208 145.1 99823 600‐37‐3; HMDB00407 1101 20.4 184.7 9.1 94 35107 isovalerylglycine Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 1961 158.1 546304 16284‐60‐9; HMDB00678 1506 7 93.4 13.3 95 33441 isobutyrylcarnitine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 1941 232.2 25518‐49‐4; HMDB00736 1513 6.7 78.5 11.7 96 35431 2‐methylbutyrylcarnitine (C5) Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2439 246.1 6426901 31023‐25‐3; HMDB00378 1469 6.9 102.2 14.8 97 31928 2‐methylbutyrylglycine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2591 160.1 193872 52320‐67‐9 ; HMDB00339 1439 16.5 58.2 3.5 98 31940 3‐methylcrotonylglycine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2580 158.1 169485 33008‐07‐0 ; HMDB00459 1240 15.3 76 5.0 99 34407 isovalerylcarnitine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2533 246.2 6426851 31023‐24‐2; HMDB00688 1154 9.6 240.1 25.0 100 35433 hydroxyisovaleroyl carnitine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 1683 262.1 99159‐87‐2; 1505 7 68.5 9.8 101 35428 tiglyl carnitine Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 2317 244.1 64191‐86‐2; HMDB02366 1511 7.6 71.4 9.4 102 37060 3‐methylglutarylcarnitine (C6) Amino acid Valine, leucine and isoleucine metabolism LC/MS Pos 1900 290.1 128145 102673‐95‐0; HMDB00552 1513 7.3 94.8 13.0 103 35437 isobutyrylglycine Amino acid Valine, leucine and isoleucine metabolism LC/MS Neg 1442 144.1 10855600 15926‐18‐8; HMDB00730 1494 6.2 81.7 13.2 104 22158 citramalate Amino acid Valine, leucine and isoleucine metabolism GC/MS 1490.7 247 1081 6236‐10‐8; C00815 HMDB00426 1517 11.2 214.4 19.1 105 31453 cysteine Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 1560.1 218 58,626,419,722 52‐90‐4;56‐89‐3; C00097 HMDB00574 1502 13.5 82.5 6.1 106 40262 S‐methylcysteine Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 1447.8 162.1 705,817,424,417 1187‐84‐4; HMDB02108 1436 21.1 196.7 9.3 107 39512 cystine Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 2015.3 145.9 595 56‐89‐3; C00491 HMDB00192 1170 33.2 213.1 6.4 108 15705 cystathionine Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 1979.6 218.1 4,083,111,834 535‐34‐2; C02291 HMDB00099 1015 29.1 258.4 8.9 109 2125 taurine Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 1616.4 254.1 11,234,068,592 107‐35‐7; C00245 HMDB00251 1329 19.3 143.4 7.4 110 1302 methionine Amino acid Cysteine, methionine, SAM, taurine metabolism LC/MS Pos 1252 150.1 69,920,876,137 63‐68‐3; C00073 HMDB00696 1515 16.1 76.9 4.8 111 21044 2‐hydroxybutyrate (AHB) Amino acid Cysteine, methionine, SAM, taurine metabolism GC/MS 1169.4 130.9 440864 3347‐90‐8; C05984 HMDB00008 1395 16.8 308 18.3 112 36808 dimethylarginine (SDMA + ADMA) Amino acid Urea cycle; arginine‐, proline‐, metabolism LC/MS Pos 812 203.2 123831 102783‐24‐4; C03626 HMDB01539 1516 8.9 53.2 6.0 113 1638 arginine Amino acid Urea cycle; arginine‐, proline‐, metabolism GC/MS 1608.5 142 5,246,487,232 1119‐34‐2; C00062 HMDB00517 1444 18.2 451.2 24.8 114 33953 N‐acetylarginine Amino acid Urea cycle; arginine‐, proline‐, metabolism LC/MS Pos 1087 217.2 674,271,615,663 155‐84‐0; C02562 HMDB04620 1516 6.2 63.9 10.3 115 1493 ornithine Amino acid Urea cycle; arginine‐, proline‐, metabolism GC/MS 1763.8 141.9 6262 3184‐13‐2; C00077 HMDB03374 1514 19.1 158.5 8.3 116 36532 urea Amino acid Urea cycle; arginine‐, proline‐, metabolism GC/MS 1303 319.1 117,616,150,869 57‐13‐6; C00086 HMDB00294 1517 13.6 26 1.9

17 of 39 117 1898 proline Amino acid Urea cycle; arginine‐, proline‐, metabolism GC/MS 1333.8 142 1,457,426,971,047 147‐85‐3; C00148 HMDB00162 1508 37.8 454.3 12.0 118 18319 5‐aminovalerate Amino acid Urea cycle; arginine‐, proline‐, metabolism GC/MS 1620.8 174 6,992,101,138 660‐88‐8; C00431 HMDB03355 1410 13.4 137 10.2 119 37431 N‐methyl proline Amino acid Urea cycle; arginine‐, proline‐, metabolism LC/MS Pos 833 130.1 557 475‐11‐6; 1208 18.1 187.2 10.3 120 22138 homocitrulline Amino acid Urea cycle; arginine‐, proline‐, metabolism LC/MS Pos 832 190.1 650,726,991,977 1190‐49‐4; C02427 HMDB00679 1511 9.1 59.4 6.5 121 27718 creatine Amino acid Creatine metabolism LC/MS Pos 758 132.1 586 57‐00‐1; C00300 HMDB00064 1517 14.3 220.8 15.4 122 513 creatinine Amino acid Creatine metabolism LC/MS Neg 1087 112.2 588 60‐27‐5; C00791 HMDB00562 1516 17.8 42 2.4 123 1577 2‐aminobutyrate Amino acid Butanoate metabolism GC/MS 1215.7 130 4,396,916,971,251 1492‐24‐6; C02261 HMDB00650 1479 11.2 121.1 10.8 124 1419 5‐methylthioadenosine (MTA) Amino acid Polyamine metabolism LC/MS Pos 2427 298.1 439176 2457‐80‐9; C00170 HMDB01173 1507 10.9 53.2 4.9 125 1408 putrescine Amino acid Polyamine metabolism GC/MS 1705.8 174 110‐60‐1; C00134 HMDB01414 1363 21 167.7 8.0 126 37496 N‐acetylputrescine Amino acid Polyamine metabolism LC/MS Pos 895 131.1 122356 18233‐70‐0; C02714 HMDB02064 1512 14.7 71.9 4.9 127 1558 4‐acetamidobutanoate Amino acid Guanidino and acetamido metabolism LC/MS Pos 1724 146.1 18189 3025‐96‐5; C02946 HMDB03681 1515 14.6 54.4 3.7 128 1494 5‐oxoproline Amino acid Glutathione metabolism LC/MS Neg 744 128.2 7405 98‐79‐3; C01879 HMDB00267 1516 4.8 44.5 9.3 129 21030 glycylglycine Peptide Dipeptide GC/MS 1447.3 258.1 154,889,711,163 556‐50‐3; C02037 HMDB11733 1460 19.3 70.8 3.7 130 22171 glycylproline Peptide Dipeptide LC/MS Pos 1115 173.1 30,136,256,993,386 704‐15‐4; HMDB00721 1466 14.5 85.5 5.9 131 15129 alanylalanine Peptide Dipeptide GC/MS 1379.5 271.1 54,603,626,992,112 1948‐31‐8; C00993 HMDB03459 1344 31.8 80.1 2.5 132 35127 pro‐hydroxy‐pro Peptide Dipeptide GC/MS 1986 267 11673055 18684‐24‐7; HMDB06695 1516 11.5 81.9 7.1 133 32394 pyroglutamylvaline Peptide Dipeptide LC/MS Neg 1755 227.2 152416 21282‐10‐0; 1376 16.7 62.4 3.7 134 41519 aspartylglycine Peptide Dipeptide GC/MS 1750.1 128 3,024,295,247,119 3790‐51‐0; 1489 20.3 78 3.8 135 1768 carnosine Peptide Dipeptide derivative LC/MS Neg 813 225.2 4,392,246,992,100 305‐84‐0; C00386 HMDB00033 910 6.9 122.6 17.8 136 15747 anserine Peptide Dipeptide derivative LC/MS Neg 1126 239.2 6,992,114,112,072 10030‐52‐1; C01262 HMDB00194 902 17.6 291.3 16.6 137 32393 gamma‐glutamylvaline Peptide gamma‐glutamyl LC/MS Pos 2040 247.2 2746‐34‐1; HMDB11172 1514 6.2 53.9 8.7 138 18369 gamma‐glutamylleucine Peptide gamma‐glutamyl LC/MS Pos 2744 261.2 151023 2566‐39‐4; HMDB11171 1325 13.3 74.8 5.6 139 34456 gamma‐glutamylisoleucine* Peptide gamma‐glutamyl LC/MS Pos 2644 261.2 HMDB11170 1513 7.1 51.9 7.3 140 33422 gamma‐glutamylphenylalanine Peptide gamma‐glutamyl LC/MS Pos 2846 295.1 111299 7432‐24‐8; HMDB00594 1236 15.4 69.1 4.5 141 2734 gamma‐glutamyltyrosine Peptide gamma‐glutamyl LC/MS Pos 2073 311.2 94340 7432‐23‐7; 1470 10.5 57.8 5.5 142 18534 glucosamine Carbohydrate Aminosugars metabolism GC/MS 1833 203 441477 66‐84‐2; C00329 HMDB01514 1461 18.8 157.1 8.4 143 33477 erythronate* Carbohydrate Aminosugars metabolism GC/MS 1546.9 292.1 2781043 88759‐55‐1; HMDB00613 1517 10.2 53.7 5.3 144 34284 N‐acetylneuraminate Carbohydrate Aminosugars metabolism GC/MS 2058 356.2 131‐48‐6; C00270 HMDB00230 1515 12.9 67.4 5.2 145 15821 fucose Carbohydrate Aminosugars metabolism GC/MS 1682.2 204 3034656 2438‐80‐4; C02095 HMDB00174 1517 11.6 84.7 7.3 146 40428 6‐sialyl‐N‐acetyllactosamine Carbohydrate Aminosugars metabolism LC/MS Neg 792.5 673.3 16212424 174757‐71‐2; HMDB06584 1516 6.7 57.4 8.6 147 577 fructose Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 1758 217 5984 57‐48‐7; C00095 HMDB00660 1517 11.3 99.8 8.8 148 1117 galactitol (dulcitol) Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 1849.2 319.1 11850 608‐66‐2; C01697 HMDB00107 1440 10.8 348.9 32.3 149 12055 galactose Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 1793.8 203.9 3037556 59‐23‐4; C01582 HMDB00143 1496 13.4 115.6 8.6 150 567 lactose Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 2134 204 6134 5965‐66‐2; C00243 HMDB00186 1513 12.6 101.6 8.1 151 15335 mannitol Carbohydrate tose, mannose, galactose, starch, and sucrose metaboLC/MS Neg 714 181.2 6251 69‐65‐8; C00392 HMDB00765 1516 5.9 234.5 39.7 152 584 mannose Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 1753.5 203.9 161658 3458‐28‐4; C00159 HMDB00169 1515 10.5 181 17.2 153 15053 sorbitol Carbohydrate tose, mannose, galactose, starch, and sucrose metabo GC/MS 1843 319.1 107428 6706‐59‐8; C00794 HMDB00247 1368 13.9 585.4 42.1 154 1519 sucrose Carbohydrate tose, mannose, galactose, starch, and sucrose metaboLC/MS Neg 875 341.2 5988 57‐50‐1; C00089 HMDB00258 1506 6.7 189 28.2 155 36649 sucralose Carbohydrate tose, mannose, galactose, starch, and sucrose metaboLC/MS Neg 3116 395 56038‐13‐2; C12285 505 16.5 177.4 10.8 156 40424 3‐sialyllactose Carbohydrate tose, mannose, galactose, starch, and sucrose metaboLC/MS Neg 725.6 632.2 123914 35890‐38‐1; HMDB00825 1516 10.7 64.6 6.0 157 20675 1,5‐anhydroglucitol (1,5‐AG) Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism LC/MS Neg 808 163.2 154‐58‐5; C07326 HMDB02712 1083 50.9 87.4 1.7 158 20488 glucose Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1866.8 217.1 79025 50‐99‐7; C00031 HMDB00122 1516 9.6 286.8 29.9 159 21049 1,6‐anhydroglucose Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1679.5 204.1 11412545 498‐07‐7; HMDB00640 1516 9.7 126 13.0 160 40264 3‐phosphoglycerate Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1754 357.1 80731‐10‐8; C00597 HMDB00807 1363 22.3 74.9 3.4 161 599 pyruvate Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1130.6 217 107735 127‐17‐3; C00022 HMDB00243 1516 11.9 81.2 6.8 162 527 lactate Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1102.8 116.9 612 79‐33‐4; C00186 HMDB00190 1517 18.2 238.6 13.1 163 15443 glucuronate Carbohydrate Glycolysis, gluconeogenesis, pyruvate metabolism GC/MS 1888 217 65041 207300‐70‐7; C00191 HMDB00127 1517 11.4 97.6 8.6 164 38075 arabitol Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1687.5 307.1 94154 7643‐75‐6; C00474 HMDB01851 1517 10.4 55.9 5.4 165 15772 ribitol Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1692.4 217 488‐81‐3; C00474 HMDB00508 1514 11 96.2 8.7 166 35854 threitol Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1513 217.1 169019 2418‐52‐2; C16884 HMDB04136 1515 9.7 70.1 7.2 167 587 gluconate Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1879.4 333 10690 527‐07‐1; C00257 HMDB00625 1508 12.6 182.1 14.5 168 35855 ribulose Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1662 306.1 79021 488‐84‐6; C00309 HMDB00621 1474 13.7 65.7 4.8 169 4966 xylitol Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1677.6 307.2 6912 87‐99‐0; C00379 HMDB02917 1516 10.4 158.3 15.2 170 575 arabinose Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1631.6 217 66308 28697‐53‐2; C00216 HMDB00646 1516 10.3 77.8 7.6 171 15835 xylose Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1723.9 204 95259 609‐06‐3; C00181 HMDB00098 1516 9.8 134.3 13.7 172 35638 xylonate Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1722 292 6602431 73686‐31‐7; C05411 1516 10.7 61.3 5.7 173 18344 xylulose Carbohydrate Nucleotide sugars, pentose metabolism GC/MS 1668 306.1 5289590 551‐84‐8; C00310 HMDB00654 1514 12.8 66.2 5.2 174 1564 citrate Energy Krebs cycle GC/MS 1763.4 273.1 311 77‐92‐9; C00158 HMDB00094 1517 8.3 100.6 12.1 175 37483 2‐methylcitrate Energy Krebs cycle GC/MS 1778 287.1 5460420 6061‐96‐7; C02225 HMDB00379 1517 12.2 58.4 4.8 176 39601 homocitrate Energy Krebs cycle GC/MS 1836.8 287.1 439459 3562‐74‐1; C01251 HMDB03518 1480 13.3 80.2 6.0 177 12025 cis‐aconitate Energy Krebs cycle LC/MS Neg 603 173.1 643757 585‐84‐2; C00417 HMDB00072 1516 11.5 45.1 3.9 178 35962 itaconate (methylenesuccinate) Energy Krebs cycle GC/MS 1570 229.1 811 97‐65‐4; C00490 HMDB02092 1429 18.4 250.4 13.6 179 12110 isocitrate Energy Krebs cycle GC/MS 1763.4 245 1198 20226‐99‐7; C00311 HMDB00193 1516 9.5 69.1 7.3 180 528 alpha‐ketoglutarate Energy Krebs cycle GC/MS 1601.5 347.1 51 305‐72‐6;328‐50‐7;22202‐68‐2; C00026 HMDB00208 1462 14.6 236.2 16.2 181 1437 succinate Energy Krebs cycle GC/MS 1348 247 1110 110‐15‐6; C00042 HMDB00254 1504 17.4 145.1 8.3 182 37058 succinylcarnitine Energy Krebs cycle LC/MS Pos 1401 262.1 1508 7.4 49 6.6 183 1643 fumarate Energy Krebs cycle GC/MS 1382.1 245 100‐17‐8; C00122 HMDB00134 1517 18.3 70.1 3.8 184 18493 mesaconate (methylfumarate) Energy Krebs cycle GC/MS 1429 184 498‐24‐8; C01732 HMDB00749 1507 14.4 85.8 6.0 185 1303 malate Energy Krebs cycle GC/MS 1502 233 525 6915‐15‐7; C00149 HMDB00156 1503 74.2 173.2 2.3 186 18367 citraconate Energy Krebs cycle, dicarboxylic acid GC/MS 1382 259 643,798,638,129 498‐23‐7; C02226 HMDB00634 1410 21.5 132 6.1 187 34425 phosphate Energy Oxidative phosphorylation LC/MS Neg 627 97.1 1061 7664‐38‐2 ; C00009 HMDB01429 1517 3.5 32.4 9.3 188 1110 arachidonate (20:4n6) Lipid Long chain fatty acid LC/MS Neg 5525 303.4 444899 506‐32‐1; C00219 HMDB01043 16 29.7 189 42103 3‐hydroxypropanoate Lipid Fatty acid, monohydroxy GC/MS 1187.1 177.1 5459847 503‐66‐2; C01013 HMDB00700 1515 11.6 152 13.1 190 31938 5‐hydroxyhexanoate Lipid Fatty acid, monohydroxy LC/MS Neg 1283 131.2 170748 44843‐89‐2 ; HMDB00525 983 6.9 72.8 10.6 191 31943 3‐hydroxysebacate Lipid Fatty acid, monohydroxy LC/MS Pos 3595 219.1 3017884 73141‐46‐5; HMDB00350 1452 16.3 82.5 5.1 192 21134 adipate Lipid Fatty acid, dicarboxylate GC/MS 1520 110.9 196 124‐04‐9; C06104 HMDB00448 1515 14.5 764 52.7 193 37253 2‐hydroxyglutarate Lipid Fatty acid, dicarboxylate GC/MS 1576 247 43 40951‐21‐1; C02630 HMDB00606 1517 11.3 121.9 10.8 194 15704 pimelate (heptanedioate) Lipid Fatty acid, dicarboxylate GC/MS 1602.9 155 385 111‐16‐0; C02656 HMDB00857 1384 30.6 98.1 3.2 195 18362 azelate (nonanedioate) Lipid Fatty acid, dicarboxylate LC/MS Neg 1322 187.2 2266 123‐99‐9; C08261 HMDB00784 1409 8.9 128.5 14.4 196 31787 ‐4‐methyl‐5‐propyl‐2‐furanpropanoa Lipid Fatty acid, dicarboxylate LC/MS Neg 2815 239.1 123979 86879‐39‐2; 744 21.5 162.7 7.6 197 32452 propionylcarnitine Lipid Fatty acid metabolism (also BCAA metabolism) LC/MS Pos 1589 218.2 107738 17298‐37‐2 ; C03017 HMDB00824 1384 6.4 173.5 27.1 198 32412 butyrylcarnitine Lipid Fatty acid metabolism (also BCAA metabolism) LC/MS Pos 2007 232.2 213144 25576‐40‐3; 675 21.6 256.3 11.9 199 35436 hexanoylglycine Lipid Fatty acid metabolism LC/MS Neg 2918 172.1 99463 24003‐67‐6; HMDB00701 1446 6.4 142 22.2 200 15500 carnitine Lipid Carnitine metabolism LC/MS Pos 702 162.2 10917 461‐05‐2; C00318 1383 4.5 245 54.4 201 32654 3‐dehydrocarnitine* Lipid Carnitine metabolism LC/MS Pos 1020 160.2 6991982 10457‐99‐5; C02636 HMDB12154 1511 8.8 159.2 18.1 202 32198 acetylcarnitine Lipid Carnitine metabolism LC/MS Pos 1203 204.2 7045767 5080‐50‐2; C02571 HMDB00201 1514 4.9 217.4 44.4 203 33936 octanoylcarnitine Lipid Carnitine metabolism LC/MS Pos 4100 288.3 123701 3671‐77‐0; C02838 HMDB00791 1410 15.1 144.8 9.6 204 22842 cholate Lipid Bile acid metabolism LC/MS Neg 5148 407.4 81‐25‐4; C00695 HMDB00619 964 9.1 215.8 23.7 205 31887 12‐dehydrocholate Lipid Bile acid metabolism LC/MS Neg 4866 405.4 94235 2458‐08‐4;2458‐08‐4 ; HMDB00400 493 12.6 92 7.3 206 1114 deoxycholate Lipid Bile acid metabolism LC/MS Neg 5268 391.4 440355 83‐44‐3; C04483 HMDB00626 290 75.9 207 31904 7‐ketodeoxycholate Lipid Bile acid metabolism LC/MS Neg 4840 405.4 188292 911‐40‐0 ; HMDB00391 589 13.3 322.8 24.3 208 32620 glycolithocholate sulfate* Lipid Bile acid metabolism LC/MS Neg 5015 512.3 72222 15324‐64‐8; C11301 HMDB02639 1211 5.2 139.8 26.9 209 36850 taurolithocholate 3‐sulfate Lipid Bile acid metabolism LC/MS Neg 5002 280.8 440071 64936‐83‐0; C03642 HMDB02580 839 8 113.7 14.2 210 32599 glycocholenate sulfate* Lipid Bile acid metabolism LC/MS Neg 4763 254.8 1273 7.6 154.6 20.3 211 32807 taurocholenate sulfate* Lipid Bile acid metabolism LC/MS Neg 4762 279.8 993 7.6 174.7 23.0 212 1497 ethanolamine Lipid Glycerolipid metabolism GC/MS 1304 174.1 141‐43‐5; C00189 HMDB00149 1512 18.5 81.9 4.4 213 15365 glycerol 3‐phosphate (G3P) Lipid Glycerolipid metabolism GC/MS 1719.7 357.1 754 29849‐82‐9; C00093 HMDB00126 1517 12.1 68.6 5.7 214 19934 myo‐inositol Lipid Inositol metabolism GC/MS 1924.9 217 87‐89‐8; C00137 HMDB00211 1517 11.9 135.7 11.4 215 37112 chiro‐inositol Lipid Inositol metabolism GC/MS 1857 318.1 643‐12‐9; 1491 10.8 274.3 25.4 216 37086 pinitol Lipid Inositol metabolism GC/MS 1773 318.1 10284‐63‐6; C03844 26 128.6 217 32379 scyllo‐inositol Lipid Inositol metabolism GC/MS 1893.8 318.2 488‐59‐5; C06153 HMDB06088 1514 13.2 96.9 7.3 218 542 3‐hydroxybutyrate (BHBA) Lipid Ketone bodies GC/MS 1203.5 116.9 441 625‐72‐9; C01089 HMDB00357 1446 31.9 688.6 21.6 219 33963 acetoacetate Lipid Ketone bodies GC/MS 1285 231.1 96 3483‐11‐2; C00164 HMDB00060 1372 23.9 926.7 38.8 220 38002 1,2‐propanediol Lipid Ketone bodies GC/MS 1041 117 57‐55‐6; C00583 HMDB01881 1504 14.9 295.9 19.9 221 17747 sphingosine Lipid Sphingolipid LC/MS Pos 5197 300.2 5353955 123‐78‐4; C00319 HMDB00252 6 10.3 222 531 3‐hydroxy‐3‐methylglutarate Lipid Mevalonate metabolism GC/MS 1598.2 247 5459993 503‐49‐1; C03761 HMDB00355 1516 13.1 86.4 6.6 223 32425 hydroisoandrosterone sulfate (DHEA Lipid Sterol/Steroid LC/MS Neg 4771 367.2 12594 651‐48‐9; C04555 HMDB01032 1262 4.9 246.2 50.2 224 31591 androsterone sulfate Lipid Sterol/Steroid LC/MS Neg 5011 369.2 2479‐86‐9 ; C00523 HMDB02759 1439 4.8 145.6 30.3 225 1769 cortisone Lipid Sterol/Steroid LC/MS Pos 4420 361.2 222,786,408,722 53‐06‐5; C00762 HMDB02802 1068 20.7 50 2.4 226 38608 tetrahydrocortisone Lipid Sterol/Steroid LC/MS Neg 5097.9 363.1 53‐05‐4; HMDB00903 651 13.1 620.9 47.4 227 37202 ndrosten‐3beta,17beta‐diol disulfate Lipid Sterol/Steroid LC/MS Neg 3735 224.3 HMDB03818 1281 5.5 239.4 43.5 228 37203 ndrosten‐3beta,17beta‐diol disulfate Lipid Sterol/Steroid LC/MS Neg 4073 224.3 HMDB03818 1333 5.1 150.7 29.5 229 37190 ha‐androstan‐3beta,17beta‐diol disu Lipid Sterol/Steroid LC/MS Neg 3872 225.2 571‐20‐0; HMDB00493 577 14.7 88.6 6.0 230 32562 pregnen‐diol disulfate* Lipid Sterol/Steroid LC/MS Neg 3886 238.3 1314 6.1 214.5 35.2 231 32827 andro steroid monosulfate 1* Lipid Sterol/Steroid LC/MS Neg 3925 385.2 C04555 HMDB02759 1477 5.6 171.6 30.6 232 32792 andro steroid monosulfate 2* Lipid Sterol/Steroid LC/MS Neg 4240 383.2 C04555 HMDB02759 1325 5.5 202.2 36.8 233 37173 21‐hydroxypregnenolone disulfate Lipid Sterol/Steroid LC/MS Neg 3931 245.3 134595 1164‐98‐3; C05485 HMDB04026 1249 5.4 107 19.8 234 40708 pregnanediol‐3‐glucuronide Lipid Sterol/Steroid LC/MS Neg 5136.9 495.4 123796 1852‐49‐9; 1491 4.9 169.1 34.5 235 3147 xanthine Nucleotide Purine metabolism, (hypo)xanthine/inosine containing GC/MS 1889.9 353 1188 69‐89‐6; C00385 HMDB00292 1506 16.8 150.8 9.0 236 15136 xanthosine Nucleotide Purine metabolism, (hypo)xanthine/inosine containingLC/MS Pos 1785 285 64959 146‐80‐5; C01762 HMDB00299 1489 16.3 57.1 3.5 237 3127 hypoxanthine Nucleotide Purine metabolism, (hypo)xanthine/inosine containing GC/MS 1759.2 265 790 68‐94‐0; C00262 HMDB00157 1512 12.3 133.4 10.8

18 of 39 238 1123 inosine Nucleotide Purine metabolism, (hypo)xanthine/inosine containingLC/MS Neg 1600 267.2 58‐63‐9; C00294 HMDB00195 1404 14.6 78 5.3 239 554 adenine Nucleotide Purine metabolism, adenine containing GC/MS 1804.2 264 190 73‐24‐5; C00147 HMDB00034 1514 16.4 54.2 3.3 240 555 adenosine Nucleotide Purine metabolism, adenine containing LC/MS Pos 1650 268.1 60961 58‐61‐7; C00212 HMDB00050 1487 10.6 72.7 6.9 241 15650 N1‐methyladenosine Nucleotide Purine metabolism, adenine containing LC/MS Pos 1356 282.1 5460178 15763‐06‐1; C02494 HMDB03331 1515 6 68.1 11.4 242 2831 osine 3',5'‐cyclic monophosphate (cA Nucleotide Purine metabolism, adenine containing LC/MS Pos 1673 330.1 274 60‐92‐4; C00575 HMDB00058 1376 21.4 57.6 2.7 243 418 guanine Nucleotide Purine metabolism, guanine containing GC/MS 1942.3 352.1 764 73‐40‐5; C00242 HMDB00132 1490 23.7 79.2 3.3 244 35114 7‐methylguanine Nucleotide Purine metabolism, guanine containing LC/MS Pos 1353 166.1 11361 578‐76‐7; C02242 HMDB00897 1515 14.5 60.9 4.2 245 22287 guanidine Nucleotide Purine metabolism, guanine containing GC/MS 1476 186.9 3520 50‐01‐1; C17349 HMDB01842 1093 39.9 66.8 1.7 246 31609 N1‐methylguanosine Nucleotide Purine metabolism, guanine containing LC/MS Pos 1850 298.1 96373 2140‐65‐0; HMDB01563 1514 10.2 55.8 5.5 247 35133 N2‐methylguanosine Nucleotide Purine metabolism, guanine containing LC/MS Pos 1950 298.1 3035422 2140‐77‐4; HMDB05862 1500 11 71.8 6.5 248 35137 N2,N2‐dimethylguanosine Nucleotide Purine metabolism, guanine containing LC/MS Pos 2157 312.1 92919 2140‐67‐2; HMDB04824 1516 8.2 52.3 6.4 249 35157 N6‐carbamoylthreonyladenosine Nucleotide Purine metabolism, guanine containing LC/MS Pos 2656 413 24719‐82‐2; 1516 5.7 48.6 8.5 250 35131 neopterin Nucleotide Purine metabolism, guanine containing LC/MS Pos 1121 254.1 440842 2009‐64‐5; C05926 HMDB00845 1476 21 133.2 6.3 251 15689 7,8‐dihydroneopterin Nucleotide Purine metabolism, guanine containing GC/MS 2114.1 494.3 65074 1218‐98‐0; C04895 HMDB02275 1247 58.8 130.8 2.2 252 1604 urate Nucleotide Purine metabolism, urate metabolism LC/MS Neg 769 167.1 69‐93‐2;120K5305; C00366 HMDB00289 1516 8.4 25.8 3.1 253 1107 allantoin Nucleotide Purine metabolism, urate metabolism GC/MS 1809.8 518.3 204 97‐59‐6; C02350 HMDB00462 1467 46.9 249.7 5.3 254 35172 orotidine Nucleotide Pyrimidine metabolism LC/MS Neg 709 287.1 92751 314‐50‐1; C01103 HMDB00788 1511 8.9 100.7 11.3 255 35130 N4‐acetylcytidine Nucleotide Pyrimidine metabolism, cytidine containing LC/MS Pos 2030 286.1 107461 3768‐18‐1; HMDB05923 1496 9.6 63.9 6.7 256 573 cytosine Nucleotide Pyrimidine metabolism, cytidine containing GC/MS 1535.5 254 597 71‐30‐7; C00380 HMDB00630 1406 48.2 145 3.0 257 1505 orotate Nucleotide Pyrimidine metabolism, orotate containing GC/MS 1709.6 254.1 967 50887‐69‐9; C00295 HMDB00226 1514 13 110.8 8.5 258 604 thymine Nucleotide Pyrimidine metabolism, thymine containing LC/MS Neg 1651 125.1 1135 65‐71‐4; C00178 HMDB00262 1300 30.6 125.3 4.1 259 1566 3‐aminoisobutyrate Nucleotide bolism, thymine containing; Valine, leucine and isoleu GC/MS 1252.2 101.9 64956 10569‐72‐9;214139‐20‐5; C05145 HMDB03911 1516 11.5 184.2 16.0 260 605 uracil Nucleotide Pyrimidine metabolism, uracil containing GC/MS 1370.4 241 1174 66‐22‐8; C00106 HMDB00300 1517 18.7 85.9 4.6 261 1559 5,6‐dihydrouracil Nucleotide Pyrimidine metabolism, uracil containing GC/MS 1505.2 243 649 504‐07‐4; C00429 HMDB00076 1476 26.8 66.3 2.5 262 33442 pseudouridine Nucleotide Pyrimidine metabolism, uracil containing LC/MS Neg 1104 243.1 1445‐07‐4; C02067 HMDB00767 1515 4.5 36.3 8.1 263 33454 gulono‐1,4‐lactone Cofactors and vitamin Ascorbate and aldarate metabolism GC/MS 1862 333.1 608 1128‐23‐0; C01040 HMDB03466 1516 14 69.4 5.0 264 1640 ascorbate (Vitamin C) Cofactors and vitamin Ascorbate and aldarate metabolism GC/MS 1850.1 332.1 134‐03‐2; C00072 HMDB00044 1454 10 290.2 29.0 265 27738 threonate Cofactors and vitamin Ascorbate and aldarate metabolism GC/MS 1560.7 292.1 151152 70753‐61‐6; C01620 HMDB00943 1517 10.7 55.8 5.2 266 37516 arabonate Cofactors and vitamin Ascorbate and aldarate metabolism GC/MS 1736 292.1 122045 7643‐75‐7; HMDB00539 1517 10.8 63.2 5.9 267 1826 folate Cofactors and vitamin Folate metabolism LC/MS Pos 2606 442 6037 59‐30‐3; C00504 HMDB00121 1 268 40173 L‐urobilin Cofactors and vitamin Hemoglobin and porphyrin metabolism LC/MS Neg 4600 593.3 5280818 34217‐90‐8; C05793 HMDB04159 128 144.1 269 2290 5‐aminolevulinate Cofactors and vitamin Hemoglobin and porphyrin metabolism GC/MS 1694.6 174 1,377,048,523 5451‐09‐2; C00430 HMDB01149 1484 19 71.4 3.8 270 32426 urobilinogen Cofactors and vitamin Hemoglobin and porphyrin metabolism LC/MS Neg 4181 591.3 26818 14684‐37‐8 ; C05790 HMDB04157 365 168.3 271 1899 quinolinate Cofactors and vitamin Nicotinate and nicotinamide metabolism GC/MS 1697.7 296.1 1066 89‐00‐9; C03722 HMDB00232 1517 16.7 108.1 6.5 272 35121 nicotinurate Cofactors and vitamin Nicotinate and nicotinamide metabolism LC/MS Neg 1461 179.1 68499 583‐08‐4; C05380 HMDB03269 16 48.2 273 32401 trigonelline (N'‐methylnicotinate)Cofactors and vitamin Nicotinate and nicotinamide metabolism LC/MS Pos 757 138.1 5570 6138‐41‐6 ; C01004 HMDB00875 1515 9.7 165 17.0 274 40469 N1‐Methyl‐2‐pyridone‐5‐carboxamidCofactors and vitamin Nicotinate and nicotinamide metabolism LC/MS Pos 1707.3 153.2 69698 701‐44‐0; C05842 HMDB04193 1516 19.4 57.4 3.0 275 1508 pantothenate Cofactors and vitamin Pantothenate and CoA metabolism LC/MS Pos 2218 220.1 6613 137‐08‐6; C00864 HMDB00210 1509 6.2 145.3 23.4 276 38276 2,3‐dihydroxyisovalerate Cofactors and vitamin Pantothenate and CoA metabolism GC/MS 1422 131 677 1756‐18‐9; C04039 1494 10.6 116.6 11.0 277 1827 riboflavin (Vitamin B2) Cofactors and vitamin Riboflavin metabolism LC/MS Pos 3111 377.2 493570 83‐88‐5; C00255 HMDB00244 1089 12.1 154.3 12.8 278 42381 gamma‐CEHC glucuronide* Cofactors and vitamin Tocopherol metabolism LC/MS Neg 2540 439.2 1489 4.5 72.4 16.1 279 31555 pyridoxate Cofactors and vitamin Vitamin B6 metabolism LC/MS Neg 2210 182.1 6723 82‐82‐6 ; C00847 HMDB00017 1516 6 267.2 44.5 280 608 pyridoxine (Vitamin B6) Cofactors and vitamin Vitamin B6 metabolism LC/MS Pos 1386 170.1 1054 58‐56‐0; C00314 HMDB02075 21 158.8 281 15753 hippurate Xenobiotics Benzoate metabolism LC/MS Neg 2136 178.1 464 495‐69‐2; C01586 HMDB00714 1517 8 116.1 14.5 282 15670 2‐methylhippurate Xenobiotics Benzoate metabolism LC/MS Pos 3125 194.1 91637 42013‐20‐7; HMDB11723 3 0.2 283 42494 3‐methyl catechol sulfate 1 Xenobiotics Benzoate metabolism LC/MS Neg 2779.2 203 1444 8.1 120.2 14.8 284 42495 3‐methyl catechol sulfate 2 Xenobiotics Benzoate metabolism LC/MS Neg 2854.5 203 812 11.6 87.6 7.6 285 42493 4‐methylcatechol sulfate Xenobiotics Benzoate metabolism LC/MS Neg 2666.4 203 1499 5.5 113.2 20.6 286 18281 2‐hydroxyhippurate (salicylurate) Xenobiotics Benzoate metabolism LC/MS Neg 2252 194.1 10253 487‐54‐7; C07588 HMDB00840 1513 8.1 342.4 42.3 287 39600 3‐hydroxyhippurate Xenobiotics Benzoate metabolism LC/MS Neg 1670 194.1 450268 1637‐75‐8; HMDB06116 1490 4.5 253.2 56.3 288 35527 4‐hydroxyhippurate Xenobiotics Benzoate metabolism LC/MS Neg 1439 194 151012 2482‐25‐9; 1516 4.2 132.1 31.5 289 22160 mandelate Xenobiotics Benzoate metabolism GC/MS 1495 178.9 1292 90‐64‐2; C01984 HMDB00703 1504 14 2419 172.8 290 1568 4‐hydroxymandelate Xenobiotics Benzoate metabolism GC/MS 1735.5 267.1 328 7198‐10‐9;184901‐84‐6; C03198 HMDB00822 1516 9.9 79.5 8.0 291 35320 catechol sulfate Xenobiotics Benzoate metabolism LC/MS Neg 1928 188.9 3083879 120‐80‐9; C00090 1514 6.7 275.7 41.1 292 15778 benzoate Xenobiotics Benzoate metabolism GC/MS 1291.5 179 243 65‐85‐0; C00180 HMDB01870 1516 23.6 155.6 6.6 293 41834 4‐hydroxybenzoate Xenobiotics Benzoate metabolism GC/MS 1623.8 223.1 1,353,702,506 99‐96‐7; C00156 HMDB00500 1516 13.4 94.5 7.1 294 36098 4‐vinylphenol sulfate Xenobiotics Benzoate metabolism LC/MS Neg 3323 199.1 6426766 2628‐17‐3; C05627 HMDB04072 1482 8.2 129.8 15.8 295 15737 glycolate (hydroxyacetate) Xenobiotics Chemical GC/MS 1119 177 3,698,251,757 79‐14‐1; C00160 HMDB00115 1517 10.4 99.6 9.6 296 22202 triethanolamine Xenobiotics Chemical GC/MS 1620 262 102‐71‐6; C06771 1515 20 553.5 27.7 297 11837 2‐hydroxyphenylacetate Xenobiotics Chemical GC/MS 1572.5 253.1 11970 614‐75‐5; C05852 HMDB00669 1460 13 55.5 4.3 298 22030 2‐hydroxyisobutyrate Xenobiotics Chemical GC/MS 1107.5 130.9 11671 594‐61‐6; HMDB00729 1516 11.7 111.7 9.5 299 38154 heptaethylene glycol Xenobiotics Chemical LC/MS Pos 2836 327.2 79718 5617‐32‐3; 1126 7.6 357.4 47.0 300 21169 3‐hydroxypyridine Xenobiotics Chemical GC/MS 1187.1 152 7971 109‐00‐2; 1505 20.2 117 5.8 301 20706 oxamate Xenobiotics Chemical GC/MS 1279.9 188 974 471‐47‐6; C01444 1465 14.9 62 4.2 302 37172 bisphenol A monosulfate Xenobiotics Chemical LC/MS Neg 4214 307 501 12.1 101.5 8.4 303 35651 ectoine Xenobiotics Chemical LC/MS Pos 832 143.1 1,260,416,993,223 96702‐03‐3; C06231 1413 30.5 93.8 3.1 304 42585 ethyl vanillate Xenobiotics Chemical GC/MS 1695.5 238.1 12038 617‐05‐0; 1094 24.1 226.8 9.4 305 40700 diglycerol Xenobiotics Chemical LC/MS Pos 1107 167.2 42953 627‐82‐7; 869 95.7 306 40506 1‐(3‐aminopropyl)‐2‐pyrrolidone Xenobiotics Chemical LC/MS Pos 1322 143.2 82111 7663‐77‐6; 1439 22 77.8 3.5 307 42638 meglumine* Xenobiotics Chemical LC/MS Pos 665 196.15 4049 6284‐40‐8; 1 308 39610 codeine Xenobiotics Drug LC/MS Pos 2201.1 300.2 76‐57‐3; HMDB04995 4 309 21320 ofloxacin Xenobiotics Drug LC/MS Pos 2608 362.1 32,885,974,583 82419‐36‐1; C07321 HMDB01929 54 273.4 310 1515 salicylate Xenobiotics Drug GC/MS 1524 267 338 69‐72‐7; C00805 HMDB01895 1260 1236.7 311 33384 salicyluric glucuronide* Xenobiotics Drug LC/MS Neg 1463 370.1 1464 45.8 281.5 6.1 312 37475 4‐acetaminophen sulfate Xenobiotics Drug LC/MS Neg 1779 230.1 83939 10066‐90‐7;32113‐41‐0; C06804 HMDB01859 1254 7.7 503.7 65.4 313 38786 N‐methyl‐acetaminophen sulfate 1* Xenobiotics Drug LC/MS Neg 2052.7 260 1095 9.7 144.6 14.9 314 12032 4‐acetamidophenol Xenobiotics Drug LC/MS Pos 2238 152.1 1983 103‐90‐2; C06804 HMDB01859 682 15.9 211.3 13.3 315 33423 p‐acetamidophenylglucuronide Xenobiotics Drug LC/MS Neg 1377 326.1 4022661 120595‐80‐4; HMDB10316 1116 6.1 459.1 75.3 316 33173 2‐hydroxyacetaminophen sulfate* Xenobiotics Drug LC/MS Neg 1703 246.1 1178 10.4 321.1 30.9 317 33178 2‐methoxyacetaminophen sulfate* Xenobiotics Drug LC/MS Neg 1977 260.1 769 11.2 531.3 47.4 318 34365 3‐(cystein‐S‐yl)acetaminophen* Xenobiotics Drug LC/MS Pos 1960 271.2 5233914 770 6.2 302.4 48.8 319 33161 methoxyacetaminophen glucuronide Xenobiotics Drug LC/MS Pos 2069 375 719 21.2 164.1 7.7 320 22115 4‐acetylphenol sulfate Xenobiotics Drug LC/MS Neg 2399 215 4684006 C00548 1352 7.9 89.1 11.3 321 22290 2‐propylpentanoate (valproate) Xenobiotics Drug GC/MS 1199.8 201.1 3121 99‐66‐1; C07185 HMDB01877 11 30.2 322 42960 clotrimazole Xenobiotics Drug LC/MS Pos 4604 277.3 2812 23593‐75‐1; D00282 HMDB01922 1 323 42984 Gemfibrozil Xenobiotics Drug LC/MS Neg 5164 249.2 3463 25812‐30‐0; D00334 10 98.8 324 35661 lidocaine Xenobiotics Drug LC/MS Pos 2761 235.2 3676 137‐58‐6; D00358 31 32.9 325 39972 metformin Xenobiotics Drug GC/MS 1746.4 284.1 4091 1115‐70‐4; C07151 HMDB01921 2 326 18037 metoprolol Xenobiotics Drug LC/MS Pos 3205 268.2 4171 56392‐17‐7; D02358 HMDB01932 698 18.7 450 24.1 327 34109 metoprolol acid metabolite* Xenobiotics Drug LC/MS Pos 2535 268.2 62936 56392‐14‐4; 977 11.8 438 37.1 328 38321 allopurinol riboside Xenobiotics Drug LC/MS Neg 1662 267.2 16220‐07‐8; 117 93.1 329 42613 famotidine Xenobiotics Drug LC/MS Neg 2639.2 336 570,216,033,255,354,000 76824‐35‐6; D00318 HMDB01919 313 123.3 330 42021 fexofenadine Xenobiotics Drug LC/MS Neg 5107.5 500.2 3348 83799‐24‐0; C06999 HMDB05030 20 70.2 331 35967 methamphetamine Xenobiotics Drug LC/MS Pos 2587 150.2 1206 33817‐09‐3; C07164 4 1 332 35432 pivaloylcarnitine Xenobiotics Drug LC/MS Pos 2370 246.2 126894 1242 14 418.3 29.9 333 42977 methylprednisolone Xenobiotics Drug LC/MS Pos 4852 375.2 6741 83‐43‐2; D00407 131 462.2 334 38327 ibuprofen acyl glucuronide Xenobiotics Drug LC/MS Neg 4806 381.1 163959 115075‐59‐7; 193 8.5 1028 120.9 335 38609 pantoprazole Xenobiotics Drug LC/MS Neg 4401.1 382.1 4679 102625‐70‐7; C11806 HMDB05017 2 336 38595 ranitidine Xenobiotics Drug LC/MS Neg 3764 313.2 3001055 66357‐59‐3; D00422 HMDB01930 154 130.3 337 37033 carbamazepine* Xenobiotics Drug LC/MS Pos 4515 237.2 2554 298‐46‐4; D00252 8 17.8 338 37020 carbamazepine 10,11‐epoxide* Xenobiotics Drug LC/MS Pos 3969 253 2555 36507‐30‐9; C07496 9 45.1 339 39625 hydrochlorothiazide Xenobiotics Drug LC/MS Neg 2041.6 296.1 3639 58‐93‐5; C07041 HMDB01928 15 31.2 340 37056 carbamazepine glucuronide* Xenobiotics Drug LC/MS Pos 4209 413.1 9 42.8 341 38686 gabapentin Xenobiotics Drug LC/MS Pos 2482 172.2 34,466,919,078 60142‐96‐3; D00332 HMDB05015 58 42.7 342 39678 benzoylecgonine Xenobiotics Drug LC/MS Pos 3100.7 290.2 442997 519‐09‐5; C10847 13 204.1 343 38658 atenolol Xenobiotics Drug LC/MS Pos 1988 267.2 2249 29122‐68‐7; D00235 HMDB01924 163 96.2 344 38619 lipitor Xenobiotics Drug LC/MS Neg 5091.5 557.3 60823 134523‐03‐8; D00887 HMDB05006 77 88.6 345 40459 escitalopram Xenobiotics Drug LC/MS Pos 3855 325.2 146570 219861‐08‐2; HMDB05028 15 37.6 346 39798 vancomycin Xenobiotics Drug LC/MS Pos 2153.1 724.9 14969 123409‐00‐7; 116 512.4 347 42990 verapamil Xenobiotics Drug LC/MS Pos 4152 455.4 2520 152‐11‐4; D02356 HMDB01850 7 46.6 348 40421 rosuvastatin Xenobiotics Drug LC/MS Neg 4511.8 480.2 446157 147098‐20‐2; D01915 16 95.7 349 39767 quinine Xenobiotics Drug LC/MS Pos 2877.2 325.3 2728270 130‐95‐0; 6 20.4 350 41725 oxypurinol Xenobiotics Drug LC/MS Pos 1530.5 153 4644 2465‐59‐0; D02365 HMDB00786 26 17.6 351 18299 N‐acetyl‐L‐cystein‐S‐yl) acetaminoph Xenobiotics Drug LC/MS Pos 2622 313.1 83967 52372‐86‐8; 929 7.3 246.2 33.7 352 15336 tartarate Xenobiotics Food component/Plant GC/MS 1626.1 292.1 875 87‐69‐4;6106‐24‐7; C00898 HMDB00956 1512 12.1 317.3 26.2 353 21151 saccharin Xenobiotics Food component/Plant LC/MS Neg 2013 182.1 5143 81‐07‐2; D01085 1266 7.6 314.5 41.4 354 32453 daidzein Xenobiotics Food component/Plant LC/MS Neg 3541 253.1 5281708 486‐66‐8 ; C10208 HMDB03312 125 151.3 355 18335 quinate Xenobiotics Food component/Plant GC/MS 1793 345.1 77‐95‐2; C00296 HMDB03072 1517 10.3 230.3 22.4 356 21156 abscisate Xenobiotics Food component/Plant LC/MS Neg 3195 263.1 5280896 14375‐45‐2; C06082 169 8.9 79.8 9.0 357 40481 dihydroferulic acid Xenobiotics Food component/Plant LC/MS Neg 1929 195.1 14340 1135‐23‐5; 588 9.6 126.6 13.2 358 35639 vanillate Xenobiotics Food component/Plant GC/MS 1728 297 8468 121‐34‐6; C06672 HMDB00484 1377 14.7 209.3 14.2

19 of 39 359 36095 thymol sulfate Xenobiotics Food component/Plant LC/MS Neg 4395 229.1 89‐83‐8; C09908 HMDB01878 1189 10.4 185.2 17.8 360 31536 N‐(2‐furoyl)glycine Xenobiotics Food component/Plant LC/MS Pos 2076 170.1 21863 5657‐19‐2;5657‐19‐2 ; HMDB00439 1513 13.6 170.8 12.6 361 37508 3,5‐dihydroxybenzoic acid Xenobiotics Food component/Plant GC/MS 1767 355.1 7424 99‐10‐5; HMDB13677 1438 14.4 133.2 9.3 362 34384 stachydrine Xenobiotics Food component/Plant LC/MS Pos 860 144.1 115244 4136‐37‐2; C10172 HMDB04827 1516 6.8 173.9 25.6 363 40479 2‐oxindole‐3‐acetate Xenobiotics Food component/Plant LC/MS Pos 3217.2 192.1 3080590 2971‐31‐5; 1451 11.4 121.3 10.6 364 38313 fucitol Xenobiotics Food component/Plant GC/MS 1742 117 3429 13074‐06‐1; 1497 14.3 62 4.3 365 38637 cinnamoylglycine Xenobiotics Food component/Plant LC/MS Neg 3139 204.1 709625 16534‐24‐0 ; 1471 5 184.5 36.9 366 40478 equol sulfate Xenobiotics Food component/Plant LC/MS Neg 3625 321.2 3 77.4 367 41948 equol glucuronide Xenobiotics Food component/Plant LC/MS Neg 3195 417.2 3 23.9 368 569 caffeine Xenobiotics Xanthine metabolism LC/MS Pos 2820 195.1 2519 58‐08‐2; C07481 HMDB01847 1043 18 111.5 6.2 369 18254 paraxanthine Xenobiotics Xanthine metabolism LC/MS Pos 2444 181.2 4687 611‐59‐6; C13747 HMDB01860 977 22.1 96 4.3 370 18392 theobromine Xenobiotics Xanthine metabolism LC/MS Pos 2136 181.1 5429 83‐67‐0; C07480 HMDB02825 1076 39.1 113.9 2.9 371 34395 1‐methylurate Xenobiotics Xanthine metabolism LC/MS Pos 1827 183.1 69726 708‐79‐2; HMDB03099 1515 13.8 59.3 4.3 372 34400 1,7‐dimethylurate Xenobiotics Xanthine metabolism LC/MS Neg 1607 195.1 91611 33868‐03‐0; C16356 HMDB11103 1363 6.2 119.1 19.2 373 34389 1‐methylxanthine Xenobiotics Xanthine metabolism LC/MS Pos 1974 167 80220 6136‐37‐4; C16358 1349 15.2 147.9 9.7 374 32445 3‐methylxanthine Xenobiotics Xanthine metabolism LC/MS Pos 1866 167.1 70639 1076‐22‐8 ; C16357 HMDB01886 1352 16.8 111.4 6.6 375 34390 7‐methylxanthine Xenobiotics Xanthine metabolism LC/MS Pos 1778 167.1 68374 552‐62‐5; C16353 HMDB01991 1193 16.3 193.6 11.9 376 34424 5‐acetylamino‐6‐amino‐3‐methylurac Xenobiotics Xanthine metabolism LC/MS Neg 1025 197.1 88299 19893‐78‐8; C16366 HMDB04400 1495 8.9 136.9 15.4 377 34401 etylamino‐6‐formylamino‐3‐methylu Xenobiotics Xanthine metabolism LC/MS Neg 930 225.1 108214 85438‐96‐6; C16365 HMDB11105 1296 7.1 149.3 21.0 378 553 cotinine Xenobiotics Tobacco metabolite LC/MS Pos 1493 177.1 854019 486‐56‐6; HMDB01046 72 33.5 379 38661 hydroxycotinine Xenobiotics Tobacco metabolite LC/MS Pos 1284.7 193.2 10219774 34834‐67‐8; HMDB01390 106 115.4 380 20699 erythritol Xenobiotics Sugar, sugar substitute, starch GC/MS 1517.5 217 149‐32‐6; C00503 HMDB02994 1517 8.8 105.8 12.0 381 39603 ethyl glucuronide Xenobiotics Detoxification metabolism LC/MS Neg 804 221.1 152226 17685‐04‐0; 50 76.5 382 32587 X ‐ 02249 LC/MS Neg 4025 267.2 1405 12.1 107.5 8.9 383 32709 X ‐ 03056 LC/MS Pos 2264 185.2 1514 4.9 55.2 11.3 384 32560 X ‐ 07765 LC/MS Neg 3705 245.1 1406 5.2 127.1 24.4 385 33782 X ‐ 10346 LC/MS Neg 3989 259.1 807 7.5 188.7 25.2 386 32578 X ‐ 11261 LC/MS Pos 3600 286.2 1511 5.5 144.1 26.2 387 32616 X ‐ 11299 LC/MS Neg 4893 507.2 924 8.6 158.2 18.4 388 32649 X ‐ 11332 LC/MS Pos 935 212.1 1489 12.4 129.7 10.5 389 32650 X ‐ 11333 LC/MS Pos 1019 212.1 1513 7.3 66.6 9.1 390 32651 X ‐ 11334 LC/MS Pos 982 259.1 1516 8.3 71.1 8.6 391 32655 X ‐ 11338 LC/MS Pos 1049 318.1 1403 23.9 77.3 3.2 392 33745 X ‐ 11357 LC/MS Neg 1327 230.2 1501 8.4 72.7 8.7 393 32740 X ‐ 11423 LC/MS Neg 1038 260.1 1514 4.3 50.2 11.7 394 32746 X ‐ 11429 LC/MS Neg 1151 245.1 1510 5.9 36.9 6.3 395 32754 X ‐ 11437 LC/MS Neg 2888 231 1357 11.9 268.3 22.5 396 32757 X ‐ 11440 LC/MS Neg 3571 246.3 1284 6.2 484.2 78.1 397 32761 X ‐ 11444 LC/MS Neg 3983 541.2 1512 3.7 117.9 31.9 398 32769 X ‐ 11452 LC/MS Neg 4109 352.1 1488 5.5 115.2 20.9 399 32787 X ‐ 11470 LC/MS Neg 4151 525.2 1421 5.9 124.7 21.1 400 32808 X ‐ 11491 LC/MS Neg 4846 567.3 1060 6.6 215.7 32.7 401 32838 X ‐ 11521 LC/MS Pos 3650 286.2 1368 6.6 153.7 23.3 402 32846 X ‐ 11529 LC/MS Neg 4845 624.3 1361 8 144.9 18.1 403 32878 X ‐ 11561 LC/MS Neg 1252 267.1 1514 6.2 62 10.0 404 32910 X ‐ 11593 LC/MS Neg 790 189.2 1516 7 43.2 6.2 405 32957 X ‐ 11640 LC/MS Neg 3776 377.1 763 7.8 187.6 24.1 406 33269 X ‐ 11662 LC/MS Pos 3172 251.1 1314 18.4 310.3 16.9 407 32996 X ‐ 11668 LC/MS Neg 1367 215.2 1472 5.3 76.2 14.4 408 33031 X ‐ 11687 LC/MS Pos 2182 384.1 1515 6 48.8 8.1 409 33172 X ‐ 11827 LC/MS Neg 1575 312.1 279 289 410 33248 X ‐ 11837 LC/MS Neg 2484 283.1 1467 6.6 171.5 26.0 411 33183 X ‐ 11838 LC/MS Neg 2314 276 1344 24.8 303.1 12.2 412 33188 X ‐ 11843 LC/MS Neg 2710 230.1 1059 6.4 310.9 48.6 413 33197 X ‐ 11852 LC/MS Neg 3324 233.1 1378 8.7 154.4 17.7 414 33290 X ‐ 11945 LC/MS Pos 1896 283.1 1516 12.7 48.7 3.8 415 33347 X ‐ 12001 LC/MS Neg 1592 229.2 1363 13.5 71.8 5.3 416 33353 X ‐ 12007 LC/MS Neg 1376 223 1505 5.4 83 15.4 417 33360 X ‐ 12014 LC/MS Neg 1998 249.1 1515 5.1 54.5 10.7 418 33370 X ‐ 12024 LC/MS Neg 1900 207.1 1516 4.5 82.8 18.4 419 33377 X ‐ 12026 LC/MS Neg 1683 180.2 1513 12.6 45.7 3.6 420 33390 X ‐ 12039 LC/MS Neg 1617 174 1510 5.6 176.8 31.6 421 33391 X ‐ 12040 LC/MS Neg 942 259 1447 9.7 121.1 12.5 422 33408 X ‐ 12056 LC/MS Pos 1129 156.2 831 17.5 213.4 12.2 423 33507 X ‐ 12092 LC/MS Pos 1142 144.2 1516 12.4 67.2 5.4 424 33508 X ‐ 12093 LC/MS Pos 1063 203.2 1504 6.9 98.8 14.3 425 33510 X ‐ 12095 LC/MS Pos 1638 153.1 1516 7.8 72.3 9.3 426 33511 X ‐ 12096 LC/MS Pos 1578 174.2 1503 8.1 81.2 10.0 427 33512 X ‐ 12097 LC/MS Pos 1526 174.2 1380 8.8 103.8 11.8 428 33515 X ‐ 12100 LC/MS Pos 1793 221.1 1507 12.1 58.4 4.8 429 33516 X ‐ 12101 LC/MS Pos 1646 164.1 1506 11.4 111.7 9.8 430 33519 X ‐ 12104 LC/MS Pos 1755 271.1 1514 6.5 54.7 8.4 431 33522 X ‐ 12107 LC/MS Pos 1786 220.1 1408 11 154.1 14.0 432 33526 X ‐ 12111 LC/MS Pos 1723 144.1 1064 19 139.8 7.4 433 33529 X ‐ 12114 LC/MS Pos 1517 365.2 1503 8.3 76.9 9.3 434 33530 X ‐ 12115 LC/MS Pos 1587 260.2 1453 7.3 101 13.8 435 33531 X ‐ 12116 LC/MS Pos 1640 286.1 1312 19.3 164 8.5 436 33532 X ‐ 12117 LC/MS Pos 1486 204.2 1494 13.4 106.3 7.9 437 33534 X ‐ 12119 LC/MS Pos 1582 208.1 1515 12.3 117 9.5 438 33538 X ‐ 12123 LC/MS Pos 1789 256.2 968 9.1 123.4 13.6 439 33540 X ‐ 12125 LC/MS Pos 1544 245.2 1506 18.3 124.1 6.8 440 33541 X ‐ 12126 LC/MS Pos 1651 326.1 1384 20.5 82.5 4.0 441 33542 X ‐ 12127 LC/MS Pos 1235 226.1 1495 22.9 180.8 7.9 442 33590 X ‐ 12170 LC/MS Pos 2534 181.1 1515 11.8 67.5 5.7 443 33609 X ‐ 12188 LC/MS Neg 2866 228.2 369 8.5 225.6 26.5 444 33610 X ‐ 12189 LC/MS Neg 1249 273.1 1512 4.8 94.6 19.7 445 35760 X ‐ 12193 LC/MS Neg 2343 201.1 1504 6.3 77.6 12.3 446 33616 X ‐ 12195 LC/MS Neg 3369 333 597 13.4 172 12.8 447 33620 X ‐ 12199 LC/MS Pos 3038 263.1 1509 10.2 85.2 8.4 448 33627 X ‐ 12206 LC/MS Neg 654 255.1 1516 7.8 43.3 5.6 449 33632 X ‐ 12211 LC/MS Neg 2582 295.2 1226 4.8 416.1 86.7 450 33633 X ‐ 12212 LC/MS Neg 3607 229.1 1316 8.7 154.6 17.8 451 33637 X ‐ 12216 LC/MS Neg 1701 228.1 1506 5.3 93.6 17.7 452 33642 X ‐ 12221 LC/MS Neg 1468 204.1 1464 10.4 106.8 10.3 453 33646 X ‐ 12225 LC/MS Pos 1009 143.2 1516 6.7 50.6 7.6 454 33652 X ‐ 12230 LC/MS Neg 3360 217.1 1393 7.5 165.6 22.1 455 33653 X ‐ 12231 LC/MS Neg 4186 352.1 1480 7.6 125.2 16.5 456 33658 X ‐ 12236 LC/MS Neg 1321 245.1 1315 37.4 128.9 3.4 457 33666 X ‐ 12244 LC/MS Pos 1147 269.2 1516 5.9 67.3 11.4 458 33675 X ‐ 12253 LC/MS Neg 1703 188.1 1479 7.9 128.6 16.3 459 33680 X ‐ 12258 LC/MS Neg 2270 230.1 1441 8.6 157 18.3 460 33683 X ‐ 12261 LC/MS Neg 1850 258.1 863 6.6 185.2 28.1 461 33684 X ‐ 12262 LC/MS Neg 2318 258.1 1344 8 134.8 16.9 462 34510 X ‐ 12267 LC/MS Neg 2968 265.2 1468 5.4 98.9 18.3 463 33829 X ‐ 12283 LC/MS Neg 2980 243.2 1346 6.1 183.1 30.0 464 33728 X ‐ 12306 LC/MS Neg 2364 247.1 937 7.6 113.8 15.0 465 33730 X ‐ 12308 LC/MS Neg 1327 210.2 1389 6.6 96.2 14.6 466 33751 X ‐ 12329 LC/MS Neg 1867 188.1 1421 5.5 149.1 27.1 467 33792 X ‐ 12364 LC/MS Pos 1800 204.1 1402 14.2 42.4 3.0 468 33807 X ‐ 12379 LC/MS Pos 3304 297.2 1298 12.9 90.6 7.0 469 33813 X ‐ 12385 LC/MS Pos 4009 220.2 1515 6.4 85.9 13.4 470 33814 X ‐ 12386 LC/MS Neg 1001 216.3 1508 5.4 88.7 16.4 471 33835 X ‐ 12407 LC/MS Neg 1902 205.1 1482 6.8 103.9 15.3 472 33836 X ‐ 12408 LC/MS Neg 1790 285.2 1088 6.5 356.7 54.9 473 33838 X ‐ 12410 LC/MS Neg 2130 274.1 1513 10.8 104.5 9.7 474 33839 X ‐ 12411 LC/MS Neg 1077 195.2 1515 13.2 77.8 5.9 475 33877 X ‐ 12435 LC/MS Neg 3174 357.2 1423 17.1 141.1 8.3 476 33904 X ‐ 12459 LC/MS Pos 4408 302.1 1430 6.2 108.6 17.5 477 33910 X ‐ 12465 LC/MS Pos 1475 248.2 1026 23.6 383.1 16.2 478 34040 X ‐ 12510 LC/MS Pos 3343 160.2 706 23.7 61.1 2.6 479 34524 X ‐ 12511 LC/MS Neg 3937 200.2 1500 6.6 111.1 16.8

20 of 39 480 34132 X ‐ 12565 LC/MS Pos 1724 254.2 1372 7.9 181.4 23.0 481 34170 X ‐ 12602 LC/MS Pos 1456 204.2 966 18.3 58.1 3.2 482 34203 X ‐ 12609 LC/MS Neg 2218 485.3 1423 5 335.1 67.0 483 34235 X ‐ 12636 LC/MS Pos 3890 259.2 1466 6.4 113.9 17.8 484 34236 X ‐ 12637 LC/MS Neg 3430 257.2 1497 5 130.4 26.1 485 34289 X ‐ 12680 LC/MS Pos 819 229.3 1474 15.9 75.2 4.7 486 34290 X ‐ 12681 LC/MS Pos 931 176.2 1492 12.5 56.3 4.5 487 34293 X ‐ 12684 LC/MS Pos 1029 108.2 780 11.5 133.3 11.6 488 34294 X ‐ 12685 LC/MS Pos 1060 154.2 1399 11.8 205.2 17.4 489 34307 X ‐ 12687 LC/MS Neg 861 355.1 1388 13 90.9 7.0 490 34297 X ‐ 12688 LC/MS Pos 1210 203.2 1496 11.2 69.9 6.2 491 34299 X ‐ 12690 LC/MS Pos 1386 346.1 1516 13.8 40.9 3.0 492 34311 X ‐ 12701 LC/MS Neg 1100 319.1 1514 6.8 99.2 14.6 493 34315 X ‐ 12705 LC/MS Neg 1245 324.1 1487 4.4 116.3 26.4 494 34316 X ‐ 12706 LC/MS Neg 1280 223 1469 6.6 101.9 15.4 495 34317 X ‐ 12707 LC/MS Neg 1260 249.1 1516 5.7 56.2 9.9 496 34318 X ‐ 12708 LC/MS Neg 1295 269 1514 8.3 70.6 8.5 497 34322 X ‐ 12712 LC/MS Neg 1690 219 1473 8.7 99.5 11.4 498 34323 X ‐ 12713 LC/MS Neg 1645 263.1 1506 6.4 48.1 7.5 499 34324 X ‐ 12714 LC/MS Neg 1640 303.1 1515 4.2 77.1 18.4 500 34325 X ‐ 12715 LC/MS Neg 1700 279.1 1516 5.9 396.9 67.3 501 34332 X ‐ 12722 LC/MS Neg 1915 249.1 926 9.2 119.3 13.0 502 34336 X ‐ 12726 LC/MS Neg 1993 233.1 1511 12.9 78.9 6.1 503 34339 X ‐ 12729 LC/MS Neg 2077 228.1 1466 7.2 145.6 20.2 504 34340 X ‐ 12730 LC/MS Neg 2095 204.1 848 9.6 123.2 12.8 505 34343 X ‐ 12733 LC/MS Neg 2079 339.2 1508 12 136.7 11.4 506 34348 X ‐ 12738 LC/MS Neg 2422 231.1 1001 8 153.9 19.2 507 34350 X ‐ 12740 LC/MS Neg 2499 287.1 1308 5.8 163 28.1 508 34351 X ‐ 12741 LC/MS Neg 2485 294.1 1244 132.2 288.6 2.2 509 34352 X ‐ 12742 LC/MS Neg 2534 241.2 1514 5.5 262.4 47.7 510 34354 X ‐ 12744 LC/MS Neg 2521 557.2 775 4.9 284.5 58.1 511 34355 X ‐ 12745 LC/MS Neg 2541 350.1 1490 10.2 99 9.7 512 34357 X ‐ 12747 LC/MS Neg 2538 507.2 1146 77.2 513 37939 X ‐ 12748 LC/MS Neg 558 320.1 1515 4.6 60.5 13.2 514 34363 X ‐ 12753 LC/MS Pos 1884 220.1 1269 11.5 135.4 11.8 515 34370 X ‐ 12760 LC/MS Pos 2001 302.2 1452 10.1 124.9 12.4 516 34477 X ‐ 12794 LC/MS Pos 859 264.2 1470 24.3 111.9 4.6 517 34485 X ‐ 12802 LC/MS Pos 2731 318.2 1512 7.9 126.6 16.0 518 34495 X ‐ 12812 LC/MS Neg 2443 208.1 1492 5.4 158.1 29.3 519 34497 X ‐ 12814 LC/MS Neg 2597 405.2 1451 6.1 115.5 18.9 520 34502 X ‐ 12819 LC/MS Neg 2702 170.1 1462 6.8 129.3 19.0 521 34503 X ‐ 12820 LC/MS Neg 2727 405.2 1449 5.5 93.4 17.0 522 34504 X ‐ 12821 LC/MS Neg 2756 383.1 1174 9 133.4 14.8 523 34505 X ‐ 12822 LC/MS Neg 2786 389.1 1513 5.5 59.8 10.9 524 40821 X ‐ 12824 LC/MS Pos 3380.1 245.2 1516 5 101.9 20.4 525 34511 X ‐ 12828 LC/MS Neg 2995 237.2 1512 6.6 74 11.2 526 34513 X ‐ 12830 LC/MS Neg 3182 372.2 1016 5.1 330.8 64.9 527 34514 X ‐ 12831 LC/MS Neg 3214 433.2 1507 6.2 116.7 18.8 528 34515 X ‐ 12832 LC/MS Neg 3247 368.1 1318 9.9 114.1 11.5 529 34516 X ‐ 12833 LC/MS Neg 3337 229.1 1362 9.9 126.3 12.8 530 40827 X ‐ 12834 LC/MS Pos 3933.7 281.2 1464 12.4 113.9 9.2 531 34522 X ‐ 12839 LC/MS Neg 3878 269.1 1473 14.2 141.6 10.0 532 34523 X ‐ 12840 LC/MS Neg 3873 338.1 1169 9.5 111.5 11.7 533 34527 X ‐ 12844 LC/MS Neg 4168 539.3 1515 3.9 97.2 24.9 534 34528 X ‐ 12845 LC/MS Neg 4234 461.3 1376 7.6 117.9 15.5 535 34529 X ‐ 12846 LC/MS Neg 4218 481.3 1507 4.6 104.4 22.7 536 34530 X ‐ 12847 LC/MS Neg 4240 227.1 1197 7.5 138.1 18.4 537 34531 X ‐ 12848 LC/MS Neg 4288 350.1 1157 7.5 111.3 14.8 538 34532 X ‐ 12849 LC/MS Neg 4726 331.2 1515 5 329.1 65.8 539 34533 X ‐ 12850 LC/MS Neg 4847 263.8 1395 7.5 185.1 24.7 540 34539 X ‐ 12855 LC/MS Pos 1404 248.2 1193 18 99.9 5.6 541 34544 X ‐ 12860 LC/MS Pos 2264 276.2 1383 10.7 333.1 31.1 542 34568 X ‐ 12884 LC/MS Neg 5903 464.4 1 543 34590 X ‐ 12906 LC/MS Neg 810 158.3 1504 9.4 62.8 6.7 544 34737 X ‐ 13046 LC/MS Neg 1459 315.2 1459 6.5 65.3 10.0 545 34798 X ‐ 13106 LC/MS Pos 1029 167.1 1472 26.4 74.4 2.8 546 34947 X ‐ 13249 LC/MS Neg 803 253.1 1486 5.8 62.9 10.8 547 34953 X ‐ 13255 LC/MS Neg 2239 367.1 787 12.6 183.4 14.6 548 35065 X ‐ 13365 LC/MS Neg 1205 161 591 8.1 139.7 17.2 549 35068 X ‐ 13368 LC/MS Neg 2228 261 1391 10.5 474.3 45.2 550 35187 X ‐ 13429 LC/MS Neg 4880 263.7 1083 6.3 160.2 25.4 551 35189 X ‐ 13431 LC/MS Pos 4230 302.3 1499 4.9 119.4 24.4 552 35225 X ‐ 13462 LC/MS Neg 1594 289.1 1515 7 74.3 10.6 553 35290 X ‐ 13529 LC/MS Pos 3088 190.1 1403 14.2 56.9 4.0 554 35331 X ‐ 13553 LC/MS Neg 1406 263 1510 5.5 56.9 10.3 555 35497 X ‐ 13688 LC/MS Neg 1720 373.1 1503 6.3 72 11.4 556 35498 X ‐ 13689 LC/MS Neg 3696 495.2 1188 20.9 138 6.6 557 35502 X ‐ 13693 LC/MS Neg 1772 375.1 1419 10.2 79.9 7.8 558 35507 X ‐ 13698 LC/MS Pos 1900 413 1470 8.6 62.8 7.3 559 35512 X ‐ 13703 LC/MS Neg 2172 367.1 624 11.1 141.8 12.8 560 35518 X ‐ 13709 LC/MS Pos 1938 302.1 1250 13.8 118.3 8.6 561 35533 X ‐ 13723 LC/MS Neg 1316 211.1 1460 5.6 144.8 25.9 562 35534 X ‐ 13724 LC/MS Neg 1330 243 1053 12.2 107.7 8.8 563 35536 X ‐ 13726 LC/MS Neg 1523 379 1458 5.9 173.3 29.4 564 35537 X ‐ 13727 LC/MS Neg 1551 232 1453 4.9 631.6 128.9 565 35539 X ‐ 13729 LC/MS Neg 1754 242 1261 10.9 70.1 6.4 566 35540 X ‐ 13730 LC/MS Neg 1806 233 1309 5.3 174.6 32.9 567 35543 X ‐ 13733 LC/MS Neg 2186 317 766 8.2 146.8 17.9 568 35544 X ‐ 13734 LC/MS Neg 2265 410.1 43 84.1 569 35545 X ‐ 13735 LC/MS Pos 1514 238.2 1285 15.1 113.5 7.5 570 35547 X ‐ 13737 LC/MS Pos 923 129.1 1513 26.3 68.4 2.6 571 35729 X ‐ 13834 LC/MS Neg 948 185.1 1497 7 82.1 11.7 572 35730 X ‐ 13835 LC/MS Neg 1014 169.1 1467 7.3 101.1 13.8 573 35731 X ‐ 13836 LC/MS Neg 987 231 1048 12.5 110.1 8.8 574 35732 X ‐ 13837 LC/MS Neg 1002 299.1 1499 6.2 95.9 15.5 575 35733 X ‐ 13838 LC/MS Neg 1012 173.1 694 6.9 140.6 20.4 576 35735 X ‐ 13840 LC/MS Neg 1130 270.1 839 5.2 134 25.8 577 35737 X ‐ 13842 LC/MS Neg 1398 303.1 852 13.6 90.5 6.7 578 35739 X ‐ 13844 LC/MS Neg 1425 209.1 1504 6.9 127 18.4 579 40102 X ‐ 13846 LC/MS Pos 2012.3 303.2 1367 8.5 155.7 18.3 580 35742 X ‐ 13847 LC/MS Neg 1426 383.1 1256 4.8 169.1 35.2 581 35743 X ‐ 13848 LC/MS Neg 1600 162 1036 7 407 58.1 582 35745 X ‐ 13850 LC/MS Neg 1986 292.1 1053 205.9 583 35759 X ‐ 13864 LC/MS Neg 2335 180.1 1344 6.9 166.3 24.1 584 35906 X ‐ 13994 LC/MS Pos 3682 329.1 38 38.2 585 35917 X ‐ 14005 LC/MS Pos 3032 505 52 50.4 586 35927 X ‐ 14015 LC/MS Neg 3539 305.1 23 68 587 35977 X ‐ 14056 LC/MS Pos 914 192.1 1477 23.9 77.7 3.3 588 35985 X ‐ 14062 LC/MS Neg 3139 283.1 743 10.1 116.1 11.5 589 36000 X ‐ 14077 LC/MS Pos 3749 263.1 792 26.7 166.4 6.2 590 36004 X ‐ 14081 LC/MS Pos 3886 346.2 952 24.5 517.1 21.1 591 36005 X ‐ 14082 LC/MS Neg 4487 511.2 1326 12.7 146.2 11.5 592 36228 X ‐ 14302 LC/MS Neg 2240 241.1 1502 7.1 79.3 11.2 593 36244 X ‐ 14318 LC/MS Pos 2509 247.1 1499 7.7 80.9 10.5 594 36278 X ‐ 14352 LC/MS Pos 2635 247.2 1513 10.1 97.3 9.6 595 36552 X ‐ 14625 LC/MS Neg 742 308.1 1472 9.5 96.6 10.2 596 36558 X ‐ 14631 LC/MS Neg 1475 246.1 1134 12.5 123.8 9.9 597 36592 X ‐ 14665 LC/MS Neg 1498 232.1 912 7.3 116.6 16.0 598 36856 X ‐ 14838 LC/MS Pos 800 141.2 1486 15.3 107.6 7.0 599 36860 X ‐ 14839 LC/MS Pos 1919 144.2 1241 23.5 104 4.4 600 37573 X ‐ 15326 LC/MS Neg 1238 503.1 917 7.2 81.5 11.3

21 of 39 601 37629 X ‐ 15382 LC/MS Pos 2540 271 1200 20 63.9 3.2 602 37709 X ‐ 15461 LC/MS Pos 2125 160.1 1484 10.1 99.9 9.9 603 37720 X ‐ 15472 LC/MS Pos 3653 204.1 761 18.6 49.1 2.6 604 37732 X ‐ 15484 LC/MS Pos 2506 271 1065 16.5 72.3 4.4 605 37734 X ‐ 15486 LC/MS Neg 3638 269.2 1516 4.4 94 21.4 606 37745 X ‐ 15497 LC/MS Neg 2877 236.1 1477 13.1 78.8 6.0 607 37751 X ‐ 15503 LC/MS Neg 1882 193.1 1516 5.4 75.2 13.9 608 37771 X ‐ 15523 LC/MS Neg 4203 309.1 1306 54.6 277.5 5.1 609 37821 X ‐ 15572 LC/MS Pos 2475 296.3 455 24 278.9 11.6 610 37850 X ‐ 15601 LC/MS Neg 2711 320.9 98 138.3 611 37855 X ‐ 15606 LC/MS Neg 3196 247 598 462.4 612 37858 X ‐ 15609 LC/MS Neg 3664 273.1 1238 9.7 508.4 52.4 613 37885 X ‐ 15636 LC/MS Neg 3814 243.1 1416 8.4 172.7 20.6 614 37895 X ‐ 15646 LC/MS Neg 2888 343.2 1390 10.1 130.7 12.9 615 37957 X ‐ 15708 LC/MS Neg 2745 589.3 1124 5.2 307.2 59.1 616 37973 X ‐ 15724 LC/MS Neg 3361 647.4 980 7.2 199.7 27.7 617 37977 X ‐ 15728 LC/MS Neg 3972 231.1 661 8.1 184.9 22.8 618 37986 X ‐ 15737 LC/MS Pos 3125 652.2 1102 7.1 289.6 40.8 619 38081 X ‐ 15824 LC/MS Pos 3114 254.1 1359 17.8 283.1 15.9 620 38082 X ‐ 15825 LC/MS Pos 2576 291.1 1341 32.8 949.5 28.9 621 38526 X ‐ 16083 LC/MS Neg 1503.6 309.2 1472 5.3 290.2 54.8 622 38530 X ‐ 16087 LC/MS Neg 4164 443.1 1499 5.8 85.9 14.8 623 38857 X ‐ 16288 LC/MS Neg 2568.2 193.1 540 23 127.4 5.5 624 38966 X ‐ 16397 LC/MS Pos 2200.8 248.1 1515 6.6 72 10.9 625 39136 X ‐ 16563 LC/MS Neg 2835.3 172.2 1263 7.3 162.5 22.3 626 40131 X ‐ 16570 LC/MS Pos 4263.5 200.2 1501 11.1 83.8 7.5 627 39153 X ‐ 16580 LC/MS Pos 1681.8 222 1499 8.6 86.1 10.0 628 39250 X ‐ 16674 LC/MS Neg 1361.5 217.2 1427 6.4 250 39.1 629 39384 X ‐ 16803 LC/MS Neg 4643.1 464.3 1309 12.1 338.1 27.9 630 39523 X ‐ 16940 LC/MS Neg 1694.1 204.9 1494 6.7 150.7 22.5 631 39530 X ‐ 16947 LC/MS Neg 3808.5 345.2 1505 5.2 282.3 54.3 632 39561 X ‐ 16975 LC/MS Pos 2291.8 255.1 1485 7.4 85 11.5 633 39641 X ‐ 17010 LC/MS Neg 3169.5 189 858 7 69.1 9.9 634 39802 X ‐ 17138 LC/MS Pos 4104.9 310.2 1452 5.5 110.4 20.1 635 39827 X ‐ 17162 LC/MS Pos 4040.8 593.4 723 15.5 253.4 16.3 636 39855 X ‐ 17185 LC/MS Neg 3069.1 215.2 1255 8.9 251.3 28.2 637 39862 X ‐ 17192 LC/MS Pos 3839.2 603.2 11 23.2 638 40097 X ‐ 17299 LC/MS Pos 1265.9 229.2 1516 9.8 77.3 7.9 639 40098 X ‐ 17300 LC/MS Pos 1475.2 116.2 864 13.1 167.4 12.8 640 40101 X ‐ 17303 LC/MS Pos 1916.8 176.1 828 8.2 127.4 15.5 641 40103 X ‐ 17305 LC/MS Pos 2157.1 296.2 1416 9.7 186.3 19.2 642 40111 X ‐ 17313 LC/MS Pos 2532.5 200.1 1234 15.9 92.2 5.8 643 40113 X ‐ 17315 LC/MS Pos 2603.4 340.2 753 17.9 102.3 5.7 644 40118 X ‐ 17320 LC/MS Pos 2715.6 184.1 1074 11.9 163.9 13.8 645 40121 X ‐ 17323 LC/MS Pos 3126.7 272.2 1425 10.2 86.2 8.5 646 40122 X ‐ 17324 LC/MS Pos 3320.5 342.3 1040 8.4 108.5 12.9 647 40123 X ‐ 17325 LC/MS Pos 3542.6 186.1 1142 12.4 138.6 11.2 648 40125 X ‐ 17327 LC/MS Pos 3601.2 257.2 1515 4.9 137.2 28.0 649 40128 X ‐ 17330 LC/MS Pos 4188.6 458.1 1516 8.3 67.1 8.1 650 40133 X ‐ 17335 LC/MS Pos 4521.7 248.2 1515 5.6 92.1 16.4 651 40135 X ‐ 17337 LC/MS Pos 4692.5 328.3 1264 5.9 200.7 34.0 652 40137 X ‐ 17339 LC/MS Pos 4740.1 543.1 1372 11 81.7 7.4 653 40138 X ‐ 17340 LC/MS Pos 4818.9 541 1390 11.1 91.6 8.3 654 40139 X ‐ 17341 LC/MS Pos 4846.4 560.2 1493 8.7 102.6 11.8 655 40141 X ‐ 17342 LC/MS Neg 1207.3 247.1 1509 7.5 59.4 7.9 656 40142 X ‐ 17343 LC/MS Neg 1312.5 191.1 1427 9 143.4 15.9 657 40153 X ‐ 17346 LC/MS Neg 1742.8 253.2 1477 5.7 129.7 22.8 658 40154 X ‐ 17347 LC/MS Neg 1851.9 219.1 1478 6.5 145.3 22.4 659 40156 X ‐ 17349 LC/MS Neg 2500.1 367.2 844 11.3 193.2 17.1 660 40157 X ‐ 17350 LC/MS Neg 2728.5 345.3 1331 4.6 233.3 50.7 661 40159 X ‐ 17351 LC/MS Neg 3391.8 243.2 1306 8.2 116.9 14.3 662 40160 X ‐ 17352 LC/MS Neg 3398.6 495.3 1469 5.7 184.7 32.4 663 40161 X ‐ 17353 LC/MS Neg 3435 507.3 1177 5 144.8 29.0 664 40162 X ‐ 17354 LC/MS Neg 3505.9 473.2 1046 5 247.3 49.5 665 40163 X ‐ 17355 LC/MS Neg 3799.5 474.2 10 666 40165 X ‐ 17357 LC/MS Neg 4095 541.3 1506 4.2 84 20.0 667 40167 X ‐ 17359 LC/MS Neg 4607.5 541.4 1516 4.1 99.2 24.2 668 40168 X ‐ 17360 LC/MS Neg 4656.6 407.4 809 7.4 393.2 53.1 669 40169 X ‐ 17361 LC/MS Neg 4849.4 331.2 1400 4.9 224.1 45.7 670 40185 X ‐ 17365 LC/MS Neg 1810.9 282.1 1481 6.2 133.9 21.6 671 40186 X ‐ 17366 LC/MS Neg 2290 336.1 1463 6.1 54.9 9.0 672 40187 X ‐ 17367 LC/MS Neg 2704.1 182.2 1276 6.3 189.7 30.1 673 40189 X ‐ 17369 LC/MS Neg 3343.4 372.2 659 7.4 124.5 16.8 674 40190 X ‐ 17370 LC/MS Neg 3513.6 368.2 1084 7.9 127.8 16.2 675 40191 X ‐ 17371 LC/MS Neg 3784.8 452.3 611 6.5 124.4 19.1 676 40218 X ‐ 17393 LC/MS Neg 1968.3 308.2 1460 5.4 173.8 32.2 677 40223 X ‐ 17398 LC/MS Neg 1778.6 391.1 1065 19.1 245.2 12.8 678 40267 X ‐ 17438 LC/MS Neg 1986.1 245.2 1420 9.3 334.5 36.0 679 40282 X ‐ 17453 LC/MS Pos 4000.6 591.3 1042 19.1 192.6 10.1 680 40291 X ‐ 17462 LC/MS Neg 4243.6 434.3 41 115.9 681 40292 X ‐ 17463 LC/MS Neg 3461.3 221.1 14 6.4 47.9 7.5 682 40553 X ‐ 17612 LC/MS Neg 4298.5 567.4 994 5 169.2 33.8 683 40597 X ‐ 17655 LC/MS Neg 4172.2 421.1 363 6 374.4 62.4 684 40770 X ‐ 17673 LC/MS Neg 1321.9 149.1 1036 7.2 200.4 27.8 685 40771 X ‐ 17674 LC/MS Neg 1439 193 1504 7.3 79.3 10.9 686 40772 X ‐ 17675 LC/MS Neg 1482.2 244 1409 12.4 86 6.9 687 40773 X ‐ 17676 LC/MS Neg 1509.5 167.1 1516 7.5 60.3 8.0 688 40774 X ‐ 17677 LC/MS Neg 1552.2 205 1411 5.2 240.9 46.3 689 40779 X ‐ 17682 LC/MS Neg 2427.8 485.2 1514 4.8 61.4 12.8 690 40782 X ‐ 17685 LC/MS Neg 2592.6 233.1 865 9.3 134.8 14.5 691 40783 X ‐ 17686 LC/MS Neg 2634.2 259.1 1358 7.1 275.8 38.8 692 40785 X ‐ 17688 LC/MS Neg 2816.5 350.1 1508 16.5 101.6 6.2 693 40789 X ‐ 17692 LC/MS Neg 3071.9 370.1 1479 6.7 149.7 22.3 694 40790 X ‐ 17693 LC/MS Neg 3121.4 267.2 1462 7.9 85.8 10.9 695 40791 X ‐ 17694 LC/MS Neg 3178.7 447.1 1217 4.1 234.2 57.1 696 40792 X ‐ 17695 LC/MS Neg 3249.8 267.2 1503 6.5 131.3 20.2 697 40795 X ‐ 17698 LC/MS Neg 3416.5 389.2 1328 9.1 133.2 14.6 698 40796 X ‐ 17699 LC/MS Neg 3503.8 216.2 1233 8.1 119.9 14.8 699 40797 X ‐ 17700 LC/MS Neg 3769.1 248.2 1423 9.9 96 9.7 700 40798 X ‐ 17701 LC/MS Neg 3802 317.1 1298 11.1 107.9 9.7 701 40799 X ‐ 17702 LC/MS Neg 3986.3 387.2 1478 4.8 102.3 21.3 702 40800 X ‐ 17703 LC/MS Neg 4288 479.3 1469 4.5 119.4 26.5 703 40801 X ‐ 17704 LC/MS Neg 4648.4 489.3 1053 6 159.5 26.6 704 40802 X ‐ 17705 LC/MS Neg 4637.4 594.4 1387 4.6 253 55.0 705 40811 X ‐ 17711 LC/MS Pos 1361.1 163.2 53 52.6 706 40815 X ‐ 17715 LC/MS Pos 2364.4 330.1 1455 13.4 57.9 4.3 707 40817 X ‐ 17717 LC/MS Pos 2593 304.2 1481 9.3 122.9 13.2 708 40823 X ‐ 17723 LC/MS Pos 3544.3 343.3 1063 16.7 102.9 6.2 709 40830 X ‐ 17730 LC/MS Pos 4185.1 326.3 1443 5.1 121.6 23.8 710 40835 X ‐ 17735 LC/MS Pos 4860 274.2 1468 7.5 101.2 13.5 711 40863 X ‐ 17761 LC/MS Pos 4910.6 274.2 1216 7 98.5 14.1 712 40867 X ‐ 17765 LC/MS Neg 4596.2 489.3 1083 7.5 133.4 17.8 713 41161 X ‐ 18059 LC/MS Neg 2863.9 175.2 1050 13.1 106.6 8.1 714 41344 X ‐ 18240 LC/MS Neg 2056.8 194.1 841 16.2 126.3 7.8 715 41418 X ‐ 18271 LC/MS Neg 2466.7 529.3 1322 5.5 290.1 52.7 716 41421 X ‐ 18273 LC/MS Neg 2615 573.3 768 8.1 260.1 32.1 717 41423 X ‐ 18275 LC/MS Neg 2751.6 617.3 922 6.8 214.3 31.5 718 41560 X ‐ 18342 LC/MS Neg 1013.8 288.1 1311 16.7 89.3 5.3 719 41563 X ‐ 18345 LC/MS Neg 1404 245.1 1278 9.1 107.3 11.8 720 41702 X ‐ 18475 LC/MS Neg 1472.7 210.2 1408 15.1 52 3.4 721 41709 X ‐ 18482 LC/MS Neg 3989 277.1 352 9.1 139 15.3

22 of 39 722 41712 X ‐ 18485 LC/MS Pos 997.5 132.1 644 34.4 393.4 11.4 723 41713 X ‐ 18486 LC/MS Pos 1162.5 247.1 1510 16.9 28.9 1.7 724 41756 X ‐ 18527 LC/MS Neg 2324.9 367 655 12.7 144.8 11.4 725 41830 X ‐ 18599 LC/MS Pos 1137 86.1 1494 211.4 147.3 0.7 726 41837 X ‐ 18603 LC/MS Neg 1402.6 142.1 1205 11.5 155 13.5 727 41840 X ‐ 18606 LC/MS Neg 3940.3 429.2 1415 16.8 84.4 5.0 728 42223 X ‐ 18838 LC/MS Neg 1777.7 338.1 1450 5.2 75.8 14.6 729 42271 X ‐ 18886 LC/MS Neg 1905.6 217.1 1419 8.7 112.3 12.9 730 42272 X ‐ 18887 LC/MS Neg 2269.3 328.2 1511 8.5 79.6 9.4 731 42314 X ‐ 18929 LC/MS Pos 3881.1 319.2 946 26.9 62.7 2.3 732 42319 X ‐ 18932 LC/MS Neg 1516.4 199.1 815 23.1 98.4 4.3 733 42322 X ‐ 18935 LC/MS Neg 1932.1 228.1 1439 4.7 107 22.8 734 42329 X ‐ 18942 LC/MS Neg 2651.4 201.1 1316 10 90.8 9.1 735 42330 X ‐ 18943 LC/MS Neg 2624.9 279.1 1171 6.1 243.5 39.9 736 42332 X ‐ 18945 LC/MS Neg 2677.1 259.2 1381 6.1 436.4 71.5 737 42335 X ‐ 18948 LC/MS Neg 2728.6 389.1 1503 10.2 78.9 7.7 738 42342 X ‐ 18955 LC/MS Neg 3503.8 335.2 1504 10.8 73.2 6.8 739 42352 X ‐ 18965 LC/MS Neg 4727 442.2 1500 7.6 118.1 15.5 740 42379 X ‐ 18981 LC/MS Pos 2445.2 284.1 1330 6.8 341.7 50.3 741 42531 X ‐ 19124 LC/MS Neg 3431.4 457.3 489 6.6 204.8 31.0 742 42547 X ‐ 19140 LC/MS Neg 3638.6 309.2 1244 27.9 302 10.8 743 42551 X ‐ 19144 LC/MS Neg 4062.4 509.3 803 16.2 70.3 4.3 744 42658 X ‐ 19182 LC/MS Neg 1639.8 198.2 959 14.6 230.7 15.8 745 42908 X ‐ 19432 LC/MS Neg 4121.4 527.5 959 11 175.8 16.0 746 42909 X ‐ 19433 LC/MS Neg 4390 525.4 1039 10.1 112.1 11.1 747 42910 X ‐ 19434 LC/MS Neg 4538 583.4 775 8.7 130.7 15.0 748 42912 X ‐ 19436 LC/MS Neg 4747 467.4 1105 18.9 67.1 3.6 749 42915 X ‐ 19439 LC/MS Neg 1994.6 326.2 1249 6.7 133.7 20.0

23 of 39 Supplemental Table 2. Characteristics of CTOT-04 kidney transplant recipients included in the analysis to develop metabolite/mRNA signatures discriminating ACR biopsies from No Rejection biopsies

Recipient Characteristics Total* Patients with Patients with No p-value (N=185) ACR Biopsies Rejection Biopsies (N=36) (N=149) Age, years Mean (SD) 46.9 (12.8) 45.1 (12.3) 47.3 (12.9) 0.20 Median 46 42 47 Min, Max 0.1, 76 24, 72 0.1, 77 Gender, N (%) Female 58 (31.4) 9 (25.0) 49 (32.9) 0.87 Male 127 (68.6) 27 (75.0) 100 (67.1) Ethnicity, N (%) Hispanic or Latino 28 (15.1) 3 (8.3) 25 (16.8) 0.18 Not Hispanic or Latino 149 (80.5) 31 (86.1) 118 (79.2) Unknown or Not Reported 8 (4.3) 2 (5.6) 6 (4.0) Race, N (%) Black or African American 68 (36.8) 14 (38.9) 54 (36.2) 0.54 White 99 (53.5) 21 (58.3) 78 (52.4) Asian 11 (6.0) 1 (2.8) 10 (6.7) American Indian or Alaska 0 0 0 Native Other 2 (1.1) 0 2 (1.1) More Than One Race 0 0 0 Unknown or Not Reported 5 (2.7) 0 5 (3.4) Induction Therapy, N (%) IL-2 Receptor Antibody 23 (13.1) 7 (19.4) 16 (11.4) 0.83 CAMPATH-1H 72 (40.9) 11 (30.6) 61 (43.6) Thymoglobulin 69 (39.2) 15 (41.7) 54 (38.6) More than one induction therapy 9 (5.1) 2 (5.6) 7 (5.0) No lnduction Therapy 3 (1.7) 1 (2.8) 2 (1.4) Missing Information, N 9 0 9 BMI Mean (SD) 28.8 (6.3) 30.1 (6.5) 28.4 (6.2) 0.09 Median 27.5 28.6 27.5 Min, Max 17, 45 20, 43 17, 45 < 18.5, N (%) 55 (31.6) 11 (33.3) 44 (31.2) 0.29 18.5 – 24.9, N (%) 53 (30.5) 7 (21.2) 46 (32.6) 25.0 – 29.9, N (%) 3 (1.7) 0 3 (2.1) ≥ 30.0, N (%) 63 (36.2) 15 (45.5) 48 (34.0) Missing 11 3 8 eGFR (mL/min)** Number of Measurements 183 37 146 Mean (SD) 30 (17) 43 (23) 0.003 Median 27 40 Min, Max 4.7, 72.3 3.4, 107.7 24 of 39 Serum Creatinine (mg/dL) Number of Measurements 183 37 146 Mean (SD) 3.7 (2.9) 2.5 (2.3) 0.0003 Median 2.8 1.9 Min, Max 1.1, 13.3 0.7, 21.2

Donor Characteristics

Age Mean (SD) 40.8 (14.4) 42.4 (11.5) 40.4 (15.1) 0.28 Median 41 40 41 Min, Max 0.4, 66 24, 65 0.4, 66 Missing 3 0 3 Gender, N (%) Female 88 (47.6) 18 (50.0) 70 (47.0) 0.54 Male 97 (52.4) 18 (50.0) 79 (53.0) Ethnicity, N (%) Hispanic or Latino 29 (15.7) 4 (11.1) 25 (16.8) 0.27 Not Hispanic or Latino 138 (74.6) 29 (80.6) 109 (58.9) Unknown or Not Reported 18 (9.7) 3 (8.3) 15 (10.1) Race, N (%) Black or African American 47 (25.4) 10 (27.8) 37 (24.8) 0.78 White 122 (66.0) 24 (66.7) 98 (65.8) Asian 5 (2.7) 1 (2.8) 4 (2.7) American Indian or Alaska 0 0 0 Native Other 1 (0.5) 0 1 (0.7) Unknown or Not Reported 10 (5.4) 1 (2.8) 9 (6.0) Source of Donor, N (%) Deceased 86 (46.5) 17 (47.2) 69 (46.3) 0.43 Living, related 53 (28.7) 9 (25.0) 44 (29.5) Living, unrelated 46 (24.9) 10 (27.8) 36 (24.2)

* Baseline characteristics are reported for 149 patients who had at least one No Rejection biopsy with a matching urine sample and 36 patients who had at least one Acute Cellular Rejection biopsy with a matching urine sample; 12 patients are in both groups. Note: one patient was 1-2 months old at the time of transplant; this patient is in both groups. ** Estimated glomerular filtration rate was measured using the modified 4-variable (serum creatinine, age, ethnicity, and gender) Modification of Diet in Renal Disease formula.

25 of 39 Supplemental Table 3 Linear regression [Mi = f(ACR)] using all samples with matching biopsy numerator denominator ncases beta p‐value p‐gain quinolinate X‐16397 248 0.89 7.3E‐09 25,031 quinolinate 4‐hydroxymandelate 248 0.88 1.1E‐08 16,954 neopterin xanthosine 234 0.90 2.0E‐08 72,220 quinolinate X‐16570 242 0.87 2.6E‐08 7,103 proline X‐13723 233 0.87 2.8E‐08 1,749 quinolinate xylitol 247 0.86 3.1E‐08 5,796 quinolinate X‐12748 248 0.85 3.5E‐08 5,259 3‐sialyllactose xanthosine 242 0.86 5.0E‐08 201,100 X‐12117 X‐13723 229 0.86 6.6E‐08 5,568 quinolinate cis‐aconitate 246 0.83 9.0E‐08 2,012 quinolinate ribulose 241 0.85 9.2E‐08 1,984 neopterin X‐16570 235 0.84 9.7E‐08 14,493 neopterin N1‐methylguanosine 238 0.83 1.2E‐07 11,361 quinolinate homovanillate (HVA) 247 0.82 1.4E‐07 1,273 quinolinate X‐13723 235 0.83 1.6E‐07 1,155 X‐12690 xanthosine 242 0.83 1.6E‐07 61,644 X‐12117 xanthosine 236 0.83 2.0E‐07 2,374 3‐sialyllactose X‐16397 247 0.77 5.9E‐07 26,062 fucitol xylitol 244 0.77 6.1E‐07 4,288 neopterin X‐12748 240 0.77 9.9E‐07 1,433 kynurenate xanthosine 241 0.78 1.0E‐06 1,262 neopterin xylitol 239 0.77 1.2E‐06 1,165 neopterin X‐16674 224 0.78 1.4E‐06 1,002 neopterin N2‐methylguanosine 236 0.77 1.6E‐06 910 X‐12690 N2‐methylguanosine 242 0.76 1.6E‐06 3,297 fucitol N2‐methylguanosine 239 0.76 1.6E‐06 3,259 X‐17366 N2‐methylguanosine 237 0.75 2.9E‐06 1,830 3‐sialyllactose X‐16570 242 0.73 3.1E‐06 1,115 fucitol 4‐hydroxymandelate 244 0.72 3.7E‐06 1,493 X‐17366 xanthosine 237 0.74 3.7E‐06 2,732 X‐18486 N2‐methylguanosine 243 0.74 3.9E‐06 1,338 N‐acetylneuraminate N2‐methylguanosine 242 0.74 4.0E‐06 1,325 X‐18486 xanthosine 242 0.73 4.7E‐06 2,172 X‐12715 4‐hydroxyphenylacetat 247 0.71 5.1E‐06 814 hydantoin‐5‐propionic acid xanthosine 223 0.74 5.4E‐06 1,894 5‐oxoproline xanthosine 242 0.70 1.1E‐05 898 X‐11945 xanthosine 242 0.70 1.3E‐05 788 X‐11491 4‐imidazoleacetate 132 1.07 1.4E‐05 3,701 fucitol isocitrate 243 0.67 1.7E‐05 3,699 fucitol ribitol 243 0.66 2.3E‐05 886 X‐11491 7‐methylxanthine 128 1.00 2.6E‐05 1,977 fucitol xylulose 244 0.64 4.5E‐05 899 proline one 245 0.63 4.9E‐05 1 fucitol ribulose 238 0.64 6.3E‐05 835 fucitol 5‐aminolevulinate 233 0.64 7.4E‐05 849 X‐11491 X‐12267 150 0.86 8.7E‐05 1,547 3‐hydroxybutyrate (BHBA) X‐13724 165 0.77 9.1E‐05 825 diglycerol X‐12747 91 1.09 9.7E‐05 947 quinolinate one 247 0.59 1.8E‐04 1 X‐12832 X‐11452 202 0.64 2.0E‐04 811 isoleucine one 244 0.58 2.1E‐04 1

26 of 39 X‐13723 one 235 ‐0.57 3.7E‐04 1 X‐12117 one 242 0.55 4.9E‐04 1 X‐16940 one 241 ‐0.55 5.8E‐04 1 leucine one 246 0.52 9.9E‐04 1 pipecolate one 239 0.52 1.1E‐03 1 paraxanthine one 158 ‐0.63 1.1E‐03 1 1,5‐anhydroglucitol (1,5‐AG) one 197 0.57 1.1E‐03 1 X‐19434 one 126 0.77 1.3E‐03 1 kynurenate one 245 0.51 1.3E‐03 1 neopterin one 240 0.51 1.4E‐03 1 myo‐inositol one 248 0.49 1.7E‐03 1 gentisate one 243 ‐0.50 1.8E‐03 1 valine one 243 0.49 1.9E‐03 1 N‐methyl‐acetaminophen sulfate 1* one 175 0.58 2.5E‐03 1 xylitol one 248 ‐0.47 2.6E‐03 1 homovanillate (HVA) one 247 ‐0.47 3.1E‐03 1 uracil one 248 ‐0.47 3.1E‐03 1 trigonelline (N'‐methylnicotinate) one 247 ‐0.46 3.3E‐03 1 X‐16570 one 242 ‐0.47 3.5E‐03 1 4‐hydroxyphenylacetate one 248 ‐0.45 4.2E‐03 1 phenylalanine one 245 0.45 4.2E‐03 1 X‐12230 one 212 ‐0.50 4.6E‐03 1 N2‐methylguanosine one 244 ‐0.45 5.3E‐03 1 4‐hydroxymandelate one 248 ‐0.44 5.5E‐03 1 xanthine one 247 ‐0.43 5.8E‐03 1 3‐(N‐acetyl‐L‐cystein‐S‐yl) acetaminophen* one 147 0.55 5.9E‐03 1 kynurenine one 245 0.43 6.4E‐03 1 phosphate one 248 0.43 6.6E‐03 1 X‐14625 one 239 0.42 8.9E‐03 1 stachydrine one 248 0.41 9.3E‐03 1 X‐17320 one 154 ‐0.51 9.9E‐03 1 acetylcarnitine one 248 0.41 9.9E‐03 1 xanthosine one 242 ‐0.42 1.0E‐02 1 X‐17366 one 240 0.40 1.2E‐02 1 2‐methoxyacetaminophen sulfate* one 124 0.53 1.2E‐02 1 X‐17185 one 194 ‐0.48 1.4E‐02 1 4‐vinylphenol sulfate one 239 ‐0.39 1.4E‐02 1 X‐12114 one 247 0.39 1.5E‐02 1 3‐sialyllactose one 248 0.38 1.5E‐02 1 X‐17370 one 161 0.48 1.6E‐02 1 X‐12221 one 234 ‐0.39 1.6E‐02 1 urate one 248 0.38 1.6E‐02 1 lysine one 246 0.38 1.7E‐02 1 3‐hydroxykynurenine one 162 0.43 1.7E‐02 1 p‐acetamidophenylglucuronide one 170 0.46 1.7E‐02 1 hydantoin‐5‐propionic acid one 229 0.38 1.7E‐02 1 1‐(3‐aminopropyl)‐2‐pyrrolidone one 235 0.38 1.7E‐02 1 citrate one 248 ‐0.37 1.8E‐02 1 X‐17735 one 232 ‐0.39 1.8E‐02 1 1‐methylxanthine one 222 ‐0.41 1.8E‐02 1 X‐18945 one 232 0.38 1.8E‐02 1 3‐(3‐hydroxyphenyl)propionate one 170 ‐0.45 1.9E‐02 1 arginine one 229 0.39 1.9E‐02 1 propionylcarnitine one 233 0.37 1.9E‐02 1

27 of 39 5‐hydroxymethyl‐2‐furoic acid one 238 ‐0.38 2.0E‐02 1 ribitol one 247 ‐0.37 2.1E‐02 1 X‐12690 one 247 0.37 2.1E‐02 1 3,4‐dihydroxyphenylacetate one 244 ‐0.37 2.1E‐02 1 X‐18887 one 246 0.36 2.1E‐02 1 2‐methylbutyrylcarnitine (C5) one 239 0.37 2.2E‐02 1 imidazole lactate one 245 ‐0.36 2.3E‐02 1 X‐16947 one 245 0.36 2.3E‐02 1 X‐11843 one 161 0.45 2.4E‐02 1 3,5‐dihydroxybenzoic acid one 233 ‐0.37 2.4E‐02 1 glucose one 248 0.35 2.5E‐02 1 X‐15503 one 248 0.35 2.6E‐02 1 X‐16674 one 232 ‐0.36 2.6E‐02 1 pantothenate one 244 ‐0.35 2.8E‐02 1 X‐12379 one 205 0.38 2.9E‐02 1 X‐12738 one 155 ‐0.44 2.9E‐02 1 1,7‐dimethylurate one 223 ‐0.37 2.9E‐02 1 X‐17369 one 93 0.55 2.9E‐02 1 3‐aminoisobutyrate one 248 ‐0.34 3.1E‐02 1 X‐12306 one 133 0.61 3.1E‐02 1 catechol sulfate one 247 ‐0.34 3.1E‐02 1 X‐18486 one 248 0.34 3.1E‐02 1 5alpha‐androstan‐3beta,17beta‐diol disulfate one 97 0.55 3.2E‐02 1 X‐12107 one 229 ‐0.37 3.3E‐02 1 X‐12039 one 246 ‐0.34 3.3E‐02 1 X‐12685 one 225 ‐0.35 3.3E‐02 1 androsterone sulfate one 233 0.34 3.4E‐02 1 X‐12364 one 228 0.36 3.4E‐02 1 homocitrulline one 244 0.34 3.4E‐02 1 X‐12840 one 183 0.39 3.5E‐02 1 3‐(cystein‐S‐yl)acetaminophen* one 133 0.47 3.5E‐02 1 X‐17398 one 178 ‐0.40 3.6E‐02 1 1,6‐anhydroglucose one 248 0.33 3.6E‐02 1 X‐13726 one 240 0.34 3.7E‐02 1 4‐acetaminophen sulfate one 203 0.36 3.9E‐02 1 hypoxanthine one 246 ‐0.33 3.9E‐02 1 X‐17350 one 211 0.35 3.9E‐02 1 azelate (nonanedioate) one 220 ‐0.34 4.1E‐02 1 xylulose one 247 ‐0.32 4.1E‐02 1 cystine one 188 0.37 4.2E‐02 1 methionine one 246 0.32 4.2E‐02 1 X‐17300 one 130 ‐0.45 4.4E‐02 1 X‐12125 one 247 0.31 4.8E‐02 1 X‐12855 one 211 0.34 4.8E‐02 1

28 of 39 Supplemental Table 4

Logistic regression [ACR = f(Mi)] using all samples with matching biopsy numerator denominator ncases OR p‐value p‐gain quinolinate X‐16397 248 2.59 1.9E‐07 1,769 neopterin xanthosine 234 2.54 6.0E‐07 3,658 3‐sialyllactose xanthosine 242 2.47 9.9E‐07 11,463 neopterin X‐16570 235 2.46 1.3E‐06 1,642 X‐12690 xanthosine 242 2.54 1.7E‐06 6,590 neopterin N1‐methylguanosine 238 2.36 1.9E‐06 1,126 fucitol xylitol 244 2.33 4.1E‐06 833 3‐sialyllactose X‐16397 247 2.24 4.5E‐06 3,778 fucitol isocitrate 243 2.02 4.9E‐05 1,324 proline one 245 1.79 1.4E‐04 1 quinolinate one 247 1.81 3.4E‐04 1 isoleucine one 244 1.77 4.5E‐04 1 X‐13723 one 235 0.52 6.1E‐04 1 X‐12117 one 242 1.70 8.5E‐04 1 X‐16940 one 241 0.57 9.0E‐04 1 pipecolate one 239 1.63 1.6E‐03 1 leucine one 246 1.62 1.7E‐03 1 kynurenate one 245 1.63 2.0E‐03 1 neopterin one 240 1.61 2.2E‐03 1 myo‐inositol one 248 1.70 2.3E‐03 1 gentisate one 243 0.59 2.4E‐03 1 valine one 243 1.63 2.5E‐03 1 xylitol one 248 0.62 3.4E‐03 1 uracil one 248 0.62 3.9E‐03 1 trigonelline (N'‐methylnicotinate) one 247 0.62 4.2E‐03 1 X‐16570 one 242 0.62 4.3E‐03 1 homovanillate (HVA) one 247 0.64 4.7E‐03 1 4‐hydroxyphenylacetate one 248 0.62 5.1E‐03 1 phenylalanine one 245 1.55 5.3E‐03 1 X‐12230 one 212 0.59 5.7E‐03 1 N2‐methylguanosine one 244 0.63 6.2E‐03 1 4‐hydroxymandelate one 248 0.64 6.6E‐03 1 xanthine one 247 0.64 6.7E‐03 1 phosphate one 248 1.64 7.2E‐03 1 kynurenine one 245 1.56 7.5E‐03 1 X‐14625 one 239 1.52 1.0E‐02 1 stachydrine one 248 1.54 1.0E‐02 1 xanthosine one 242 0.65 1.1E‐02 1 acetylcarnitine one 248 1.51 1.1E‐02 1 X‐17366 one 240 1.49 1.4E‐02 1 4‐vinylphenol sulfate one 239 0.67 1.5E‐02 1 X‐12114 one 247 1.49 1.6E‐02 1 3‐sialyllactose one 248 1.44 1.7E‐02 1 X‐12221 one 234 0.67 1.7E‐02 1 urate one 248 1.49 1.8E‐02 1 lysine one 246 1.46 1.8E‐02 1 hydantoin‐5‐propionic acid one 229 1.44 1.9E‐02 1 1‐(3‐aminopropyl)‐2‐pyrrolidone one 235 1.45 1.9E‐02 1 citrate one 248 0.69 2.0E‐02 1 X‐17735 one 232 0.66 2.0E‐02 1 1‐methylxanthine one 222 0.65 2.0E‐02 1

29 of 39 X‐18945 one 232 1.47 2.0E‐02 1 arginine one 229 1.46 2.1E‐02 1 propionylcarnitine one 233 1.47 2.1E‐02 1 5‐hydroxymethyl‐2‐furoic acid one 238 0.67 2.1E‐02 1 X‐18887 one 246 1.46 2.2E‐02 1 ribitol one 247 0.70 2.3E‐02 1 X‐12690 one 247 1.44 2.3E‐02 1 3,4‐dihydroxyphenylacetate one 244 0.70 2.4E‐02 1 2‐methylbutyrylcarnitine (C5) one 239 1.48 2.4E‐02 1 imidazole lactate one 245 0.69 2.4E‐02 1 X‐16947 one 245 1.43 2.5E‐02 1 3,5‐dihydroxybenzoic acid one 233 0.66 2.6E‐02 1 X‐15503 one 248 1.43 2.7E‐02 1 glucose one 248 1.40 2.7E‐02 1 X‐16674 one 232 0.68 2.7E‐02 1 pantothenate one 244 0.70 2.9E‐02 1 1,7‐dimethylurate one 223 0.70 3.1E‐02 1 X‐12379 one 205 1.45 3.1E‐02 1 3‐aminoisobutyrate one 248 0.70 3.2E‐02 1 X‐18486 one 248 1.44 3.3E‐02 1 catechol sulfate one 247 0.72 3.4E‐02 1 X‐12107 one 229 0.67 3.4E‐02 1 X‐12039 one 246 0.71 3.5E‐02 1 X‐12685 one 225 0.68 3.5E‐02 1 X‐12364 one 228 1.46 3.5E‐02 1 androsterone sulfate one 233 1.42 3.6E‐02 1 homocitrulline one 244 1.40 3.6E‐02 1 1,6‐anhydroglucose one 248 1.40 3.8E‐02 1 X‐13726 one 240 1.42 3.9E‐02 1 hypoxanthine one 246 0.71 4.0E‐02 1 4‐acetaminophen sulfate one 203 1.44 4.1E‐02 1 X‐17350 one 211 1.41 4.1E‐02 1 azelate (nonanedioate) one 220 0.69 4.3E‐02 1 xylulose one 247 0.73 4.3E‐02 1 methionine one 246 1.36 4.5E‐02 1 X‐12125 one 247 1.36 4.9E‐02 1

30 of 39 Supplemental Table 5

Linear regression [Mi = f(ACR)] using all samples with matching biopsy and available mRNA signature numerator denominator ncases beta p‐value p‐gain quinolinate 4‐hydroxymandelate 206 0.96 6.7E‐09 59,573 quinolinate xylitol 205 0.93 2.3E‐08 17,578 3‐sialyllactose xanthosine 201 0.93 4.5E‐08 191,212 proline 4‐hydroxyphenylacetat 205 0.90 5.1E‐08 823 quinolinate X‐16397 206 0.89 8.2E‐08 4,854 neopterin xanthosine 195 0.92 1.0E‐07 50,721 quinolinate xylulose 206 0.86 2.2E‐07 1,773 quinolinate homovanillate (HVA) 205 0.87 2.3E‐07 1,692 pipecolate 4‐hydroxyphenylacetat 199 0.87 2.4E‐07 1,763 X‐12117 xanthosine 196 0.89 2.7E‐07 2,971 neopterin N1‐methylguanosine 198 0.87 3.0E‐07 17,759 quinolinate arabitol 206 0.85 3.4E‐07 1,158 X‐17366 xanthosine 197 0.88 3.5E‐07 24,658 X‐12117 X‐13723 193 0.87 4.0E‐07 2,048 X‐18486 xanthosine 201 0.87 4.7E‐07 18,221 quinolinate X‐16570 203 0.85 4.9E‐07 802 neopterin 4‐hydroxyphenylacetat 200 0.85 5.3E‐07 796 pipecolate xanthosine 194 0.87 6.2E‐07 912 X‐12117 xylonate 201 0.84 6.3E‐07 1,292 kynurenate xanthosine 201 0.85 7.1E‐07 12,163 X‐12117 galactose 200 0.84 7.2E‐07 1,126 X‐12114 4‐hydroxymandelate 204 0.82 8.2E‐07 1,364 X‐12117 N2‐methylguanosine 195 0.86 8.4E‐07 961 neopterin xylitol 199 0.84 8.4E‐07 816 X‐12117 4‐hydroxymandelate 201 0.83 8.8E‐07 917 X‐12117 aconate (methylfumar 198 0.83 1.1E‐06 751 2‐methylbutyrylcarnitine (C5) xanthine 199 0.82 1.3E‐06 865 X‐11491 X‐13723 126 1.09 1.3E‐06 929 neopterin X‐16570 197 0.82 1.5E‐06 3,384 X‐12690 xanthosine 201 0.82 1.8E‐06 4,760 neopterin X‐16674 186 0.83 2.0E‐06 2,612 3‐sialyllactose X‐16397 206 0.79 2.3E‐06 11,237 methionine xanthosine 199 0.82 2.4E‐06 1,764 X‐17366 N2‐methylguanosine 197 0.82 2.4E‐06 3,487 neopterin aconate (methylfumar 197 0.80 2.7E‐06 1,928 stachydrine nelline (N'‐methylnicot 206 0.78 2.8E‐06 770 X‐11491 4‐imidazoleacetate 114 1.21 2.8E‐06 16,735 neopterin X‐12748 200 0.79 3.5E‐06 1,514 X‐18486 N2‐methylguanosine 201 0.80 4.1E‐06 2,026 neopterin trans‐urocanate 199 0.78 4.1E‐06 1,268 neopterin hylarginine (SDMA + A 200 0.77 5.2E‐06 1,005 X‐11491 X‐12267 126 1.02 1.0E‐05 4,717 3‐sialyllactose X‐12748 205 0.73 1.2E‐05 2,059 X‐11491 7‐methylxanthine 109 1.05 1.6E‐05 2,946 glucose imelate (heptanedioat 185 0.77 1.7E‐05 784 3‐sialyllactose aconate (methylfumar 203 0.71 2.4E‐05 1,055 3‐sialyllactose N1‐methylguanosine 205 0.71 2.6E‐05 985 5‐oxoproline aconate (methylfumar 203 0.71 2.8E‐05 1,303 pregnen‐diol disulfate* droxypregnenolone dis 163 0.83 3.5E‐05 3,508 proline one 204 0.69 4.2E‐05 1 leucine one 203 0.69 4.4E‐05 1

31 of 39 X‐11491 X‐12712 128 0.94 5.1E‐05 919 X‐11491 X‐17349 66 1.16 5.2E‐05 899 X‐11491 1‐methylxanthine 120 1.00 5.4E‐05 868 X‐12637 X‐17699 165 0.79 5.5E‐05 994 1,5‐anhydroglucitol (1,5‐AG) one 165 0.74 7.0E‐05 1 X‐17327 X‐12819 201 0.66 1.1E‐04 1,256 X‐12116 galactose 184 0.70 1.4E‐04 882 3‐hydroxybutyrate (BHBA) X‐13724 136 0.80 1.6E‐04 759 X‐12742 X‐12819 202 0.64 1.8E‐04 771 fucitol allo‐threonine 203 0.63 2.1E‐04 935 isoleucine one 202 0.62 2.8E‐04 1 N‐methyl‐acetaminophen sulfate 1* one 145 0.74 3.4E‐04 1 fucitol arabitol 204 0.61 3.5E‐04 779 quinolinate one 205 0.60 4.0E‐04 1 4‐hydroxyphenylacetate one 206 ‐0.60 4.3E‐04 1 X‐16940 one 201 ‐0.60 5.5E‐04 1 pipecolate one 199 0.59 5.6E‐04 1 xylitol one 206 ‐0.58 6.9E‐04 1 X‐12117 one 201 0.58 8.1E‐04 1 homovanillate (HVA) one 205 ‐0.57 8.2E‐04 1 3‐(N‐acetyl‐L‐cystein‐S‐yl) acetaminophen* one 119 0.71 1.1E‐03 1 gentisate one 201 ‐0.57 1.1E‐03 1 4‐hydroxymandelate one 205 ‐0.55 1.1E‐03 1 xanthine one 206 ‐0.55 1.1E‐03 1 X‐13723 one 198 ‐0.56 1.3E‐03 1 uracil one 206 ‐0.55 1.3E‐03 1 phenylalanine one 204 0.53 1.8E‐03 1 stachydrine one 206 0.52 2.2E‐03 1 valine one 203 0.52 2.4E‐03 1 trigonelline (N'‐methylnicotinate) one 206 ‐0.51 2.5E‐03 1 X‐12855 one 174 0.56 2.8E‐03 1 lysine one 204 0.51 2.8E‐03 1 X‐17320 one 128 ‐0.63 3.1E‐03 1 paraxanthine one 131 ‐0.62 3.3E‐03 1 X‐12230 one 177 ‐0.56 3.4E‐03 1 3,4‐dihydroxyphenylacetate one 203 ‐0.50 3.7E‐03 1 2‐methoxyacetaminophen sulfate* one 101 0.67 3.8E‐03 1 methionine one 204 0.49 4.2E‐03 1 kynurenine one 204 0.48 4.8E‐03 1 X‐16570 one 203 ‐0.48 5.1E‐03 1 X‐19434 one 110 0.71 5.1E‐03 1 neopterin one 200 0.48 5.2E‐03 1 X‐12221 one 195 ‐0.48 6.0E‐03 1 p‐acetamidophenylglucuronide one 137 0.55 7.8E‐03 1 5‐hydroxymethyl‐2‐furoic acid one 199 ‐0.46 8.2E‐03 1 N2‐methylguanosine one 202 ‐0.46 8.4E‐03 1 acetylcarnitine one 206 0.45 8.6E‐03 1 xanthosine one 201 ‐0.46 8.6E‐03 1 kynurenate one 204 0.45 9.0E‐03 1 X‐17366 one 200 0.45 9.3E‐03 1 ornithine one 204 0.44 9.4E‐03 1 X‐14625 one 200 0.45 9.6E‐03 1 propionylcarnitine one 193 0.45 1.0E‐02 1 2‐methylbutyrylcarnitine (C5) one 198 0.45 1.0E‐02 1

32 of 39 isobutyrylglycine one 200 ‐0.45 1.1E‐02 1 myo‐inositol one 206 0.43 1.2E‐02 1 imidazole lactate one 203 ‐0.43 1.2E‐02 1 isovalerylglycine one 203 ‐0.43 1.3E‐02 1 urate one 206 0.42 1.3E‐02 1 glucose one 206 0.42 1.3E‐02 1 mannose one 206 0.42 1.4E‐02 1 5alpha‐androstan‐3beta,17beta‐diol disulfate one 83 0.68 1.4E‐02 1 3‐hydroxykynurenine one 133 0.48 1.5E‐02 1 X‐12039 one 205 ‐0.42 1.5E‐02 1 isocitrate one 206 ‐0.41 1.6E‐02 1 X‐12107 one 187 ‐0.46 1.6E‐02 1 X‐11843 one 130 0.53 1.6E‐02 1 ribitol one 205 ‐0.41 1.6E‐02 1 arginine one 191 0.42 1.8E‐02 1 4‐hydroxybenzoate one 206 ‐0.40 1.8E‐02 1 3‐(cystein‐S‐yl)acetaminophen* one 106 0.57 1.8E‐02 1 X‐17369 one 74 0.65 2.0E‐02 1 X‐12685 one 187 ‐0.42 2.0E‐02 1 X‐17735 one 194 ‐0.42 2.0E‐02 1 3‐aminoisobutyrate one 206 ‐0.40 2.0E‐02 1 X‐12193 one 204 ‐0.40 2.1E‐02 1 X‐16674 one 192 ‐0.41 2.1E‐02 1 allantoin one 197 0.40 2.2E‐02 1 X‐17185 one 161 ‐0.49 2.2E‐02 1 X‐12114 one 204 0.39 2.3E‐02 1 hydantoin‐5‐propionic acid one 190 0.40 2.3E‐02 1 xylulose one 205 ‐0.39 2.4E‐02 1 lactate one 202 0.39 2.4E‐02 1 1,7‐dimethylurate one 187 ‐0.41 2.5E‐02 1 X‐12849 one 206 ‐0.38 2.5E‐02 1 trans‐urocanate one 204 ‐0.38 2.5E‐02 1 tyrosine one 204 0.38 2.5E‐02 1 3‐sialyllactose one 206 0.38 2.6E‐02 1 3‐(3‐hydroxyphenyl)propionate one 141 ‐0.46 2.6E‐02 1 X‐15461 one 200 ‐0.39 2.6E‐02 1 citrate one 206 ‐0.38 2.6E‐02 1 cystine one 164 0.42 2.8E‐02 1 pantothenate one 203 ‐0.38 2.8E‐02 1 X‐18965 one 201 ‐0.38 2.9E‐02 1 X‐12726 one 204 ‐0.38 3.1E‐02 1 hypoxanthine one 206 ‐0.37 3.2E‐02 1 X‐17335 one 205 ‐0.37 3.3E‐02 1 X‐18486 one 206 0.36 3.4E‐02 1 3,5‐dihydroxybenzoic acid one 194 ‐0.38 3.4E‐02 1 X‐15484 one 142 0.43 3.4E‐02 1 X‐18887 one 205 0.36 3.5E‐02 1 X‐11668 one 199 ‐0.37 3.5E‐02 1 X‐13850 one 139 0.43 3.6E‐02 1 N‐(2‐furoyl)glycine one 206 ‐0.36 3.6E‐02 1 guanine one 198 ‐0.37 3.6E‐02 1 mesaconate (methylfumarate) one 202 ‐0.36 3.7E‐02 1 glutamine one 204 0.36 3.7E‐02 1 X‐17361 one 192 ‐0.37 3.8E‐02 1

33 of 39 taurine one 193 ‐0.38 4.2E‐02 1 X‐17398 one 153 ‐0.42 4.2E‐02 1 X‐12747 one 132 ‐0.43 4.4E‐02 1 4‐acetaminophen sulfate one 169 0.38 4.5E‐02 1 X‐12738 one 128 ‐0.46 4.5E‐02 1 X‐18945 one 195 0.35 4.5E‐02 1 2‐hydroxyacetaminophen sulfate* one 143 0.40 4.6E‐02 1 phosphate one 206 0.34 4.7E‐02 1 X‐11491 one 127 0.48 4.7E‐02 1 dimethylarginine (SDMA + ADMA) one 206 ‐0.34 4.8E‐02 1 catechol sulfate one 206 ‐0.34 5.0E‐02 1

34 of 39 Supplemental Table 6

Logistic regression [ACR = f(Mi)] using all samples with matching biopsy and available mRNA signature numerator denominator ncases OR p‐value p‐gain quinolinate 4‐hydroxymandelate 206 2.67 7.3E‐07 1,139 quinolinate xylitol 205 2.56 9.1E‐07 909 3‐sialyllactose xanthosine 201 2.74 1.7E‐06 5,814 neopterin xanthosine 195 2.65 2.7E‐06 2,631 X‐17366 xanthosine 197 2.59 4.5E‐06 2,194 neopterin N1‐methylguanosine 198 2.47 5.4E‐06 1,315 X‐18486 xanthosine 201 2.61 6.1E‐06 1,626 kynurenate xanthosine 201 2.62 7.1E‐06 1,392 X‐12690 xanthosine 201 2.51 1.2E‐05 818 3‐sialyllactose X‐16397 206 2.31 1.9E‐05 1,499 pregnen‐diol disulfate* droxypregnenolone dis 163 2.39 1.2E‐04 992 proline one 204 1.89 1.5E‐04 1 leucine one 203 1.97 1.8E‐04 1 1,5‐anhydroglucitol (1,5‐AG) one 165 2.30 2.1E‐04 1 isoleucine one 202 1.87 5.6E‐04 1 4‐hydroxyphenylacetate one 206 0.53 7.7E‐04 1 quinolinate one 205 1.79 8.3E‐04 1 N‐methyl‐acetaminophen sulfate 1* one 145 2.07 8.8E‐04 1 X‐16940 one 201 0.55 9.3E‐04 1 pipecolate one 199 1.77 1.0E‐03 1 xylitol one 206 0.55 1.2E‐03 1 X‐12117 one 201 1.72 1.4E‐03 1 gentisate one 201 0.55 1.6E‐03 1 xanthine one 206 0.55 1.7E‐03 1 X‐13723 one 198 0.52 1.9E‐03 1 uracil one 206 0.57 1.9E‐03 1 4‐hydroxymandelate one 205 0.58 1.9E‐03 1 homovanillate (HVA) one 205 0.58 2.0E‐03 1 3‐(N‐acetyl‐L‐cystein‐S‐yl) acetaminophen* one 119 2.25 2.3E‐03 1 phenylalanine one 204 1.67 2.8E‐03 1 stachydrine one 206 1.77 2.8E‐03 1 valine one 203 1.67 3.3E‐03 1 trigonelline (N'‐methylnicotinate) one 206 0.59 3.5E‐03 1 lysine one 204 1.67 3.8E‐03 1 X‐12855 one 174 1.76 4.1E‐03 1 X‐17320 one 128 0.49 4.5E‐03 1 X‐12230 one 177 0.54 4.5E‐03 1 paraxanthine one 131 0.51 5.1E‐03 1 methionine one 204 1.61 5.8E‐03 1 kynurenine one 204 1.67 5.8E‐03 1 3,4‐dihydroxyphenylacetate one 203 0.62 6.1E‐03 1 X‐16570 one 203 0.60 6.2E‐03 1 2‐methoxyacetaminophen sulfate* one 101 2.09 7.1E‐03 1 neopterin one 200 1.57 7.1E‐03 1 X‐12221 one 195 0.61 7.3E‐03 1 X‐19434 one 110 2.07 8.2E‐03 1 5‐hydroxymethyl‐2‐furoic acid one 199 0.61 9.6E‐03 1 N2‐methylguanosine one 202 0.62 9.8E‐03 1 xanthosine one 201 0.61 9.9E‐03 1 p‐acetamidophenylglucuronide one 137 1.83 1.0E‐02 1

35 of 39 acetylcarnitine one 206 1.57 1.0E‐02 1 X‐17366 one 200 1.58 1.1E‐02 1 kynurenate one 204 1.55 1.1E‐02 1 ornithine one 204 1.54 1.1E‐02 1 X‐14625 one 200 1.57 1.1E‐02 1 propionylcarnitine one 193 1.59 1.2E‐02 1 2‐methylbutyrylcarnitine (C5) one 198 1.62 1.2E‐02 1 isobutyrylglycine one 200 0.63 1.2E‐02 1 myo‐inositol one 206 1.58 1.3E‐02 1 imidazole lactate one 203 0.64 1.4E‐02 1 urate one 206 1.58 1.4E‐02 1 isovalerylglycine one 203 0.64 1.4E‐02 1 glucose one 206 1.50 1.5E‐02 1 mannose one 206 1.50 1.6E‐02 1 X‐12039 one 205 0.65 1.7E‐02 1 X‐12107 one 187 0.61 1.8E‐02 1 3‐hydroxykynurenine one 133 1.60 1.8E‐02 1 ribitol one 205 0.67 1.9E‐02 1 X‐11843 one 130 1.70 1.9E‐02 1 4‐hydroxybenzoate one 206 0.63 2.0E‐02 1 arginine one 191 1.51 2.0E‐02 1 5alpha‐androstan‐3beta,17beta‐diol disulfate one 83 1.94 2.1E‐02 1 isocitrate one 206 0.68 2.1E‐02 1 X‐12685 one 187 0.62 2.2E‐02 1 3‐(cystein‐S‐yl)acetaminophen* one 106 1.80 2.2E‐02 1 3‐aminoisobutyrate one 206 0.66 2.2E‐02 1 X‐17735 one 194 0.64 2.2E‐02 1 X‐12193 one 204 0.66 2.3E‐02 1 X‐16674 one 192 0.65 2.3E‐02 1 allantoin one 197 1.52 2.4E‐02 1 X‐17185 one 161 0.54 2.4E‐02 1 X‐12114 one 204 1.51 2.5E‐02 1 X‐17369 one 74 2.10 2.5E‐02 1 hydantoin‐5‐propionic acid one 190 1.47 2.6E‐02 1 trans‐urocanate one 204 0.68 2.7E‐02 1 xylulose one 205 0.68 2.7E‐02 1 1,7‐dimethylurate one 187 0.67 2.7E‐02 1 X‐12849 one 206 0.69 2.8E‐02 1 lactate one 202 1.44 2.8E‐02 1 tyrosine one 204 1.47 2.8E‐02 1 X‐15461 one 200 0.67 2.8E‐02 1 3‐(3‐hydroxyphenyl)propionate one 141 0.62 2.8E‐02 1 3‐sialyllactose one 206 1.44 2.8E‐02 1 citrate one 206 0.68 2.8E‐02 1 pantothenate one 203 0.68 3.1E‐02 1 cystine one 164 1.54 3.1E‐02 1 X‐18965 one 201 0.68 3.1E‐02 1 X‐12726 one 204 0.68 3.4E‐02 1 hypoxanthine one 206 0.68 3.4E‐02 1 X‐17335 one 205 0.69 3.5E‐02 1 X‐18486 one 206 1.48 3.6E‐02 1 3,5‐dihydroxybenzoic acid one 194 0.65 3.6E‐02 1 X‐15484 one 142 1.58 3.7E‐02 1 X‐18887 one 205 1.45 3.7E‐02 1

36 of 39 N‐(2‐furoyl)glycine one 206 0.69 3.8E‐02 1 X‐11668 one 199 0.70 3.8E‐02 1 X‐13850 one 139 1.53 3.9E‐02 1 guanine one 198 0.69 3.9E‐02 1 glutamine one 204 1.44 3.9E‐02 1 mesaconate (methylfumarate) one 202 0.69 3.9E‐02 1 X‐17361 one 192 0.68 4.0E‐02 1 taurine one 193 0.69 4.4E‐02 1 X‐17398 one 153 0.62 4.4E‐02 1 4‐acetaminophen sulfate one 169 1.45 4.7E‐02 1 X‐18945 one 195 1.42 4.8E‐02 1 X‐12738 one 128 0.62 4.9E‐02 1 2‐hydroxyacetaminophen sulfate* one 143 1.52 4.9E‐02 1 phosphate one 206 1.43 4.9E‐02 1 X‐12747 one 132 0.67 5.0E‐02 1

37 of 39 Supplemental Table 7

Logistic regression [ACR = f(Mi, RNA_signature)] using all samples with matching biopsy and available mRNA signature ‐ ‐ ‐ ‐ Metabolite ‐ ‐ ‐ ‐ mRNA signature numerator denominator ncases OR p‐value p‐gain OR p‐value 3‐sialyllactose xanthosine 200 3.81 1.6E‐06 3,702 3.25 3.8E‐08 X‐12690 xanthosine 200 3.72 4.3E‐06 1,024 3.17 1.0E‐08 X‐12715 4‐hydroxyphenylacetat 205 3.02 1.0E‐05 1,199 3.42 4.7E‐09 X‐12225 homovanillate (HVA) 202 3.13 1.1E‐05 1,255 3.38 2.5E‐09 X‐11334 xanthosine 200 2.81 3.8E‐05 1,189 3.17 3.0E‐09 X‐11945 xanthosine 200 2.95 3.9E‐05 784 3.02 1.0E‐08 X‐12117 one 200 2.66 6.9E‐05 1 3.25 1.7E‐09 leucine one 202 2.14 5.4E‐04 1 2.81 7.2E‐09 quinolinate one 204 2.16 7.9E‐04 1 2.91 1.8E‐09 stachydrine one 205 2.21 9.4E‐04 1 3.00 1.7E‐09 X‐16940 one 200 0.50 1.3E‐03 1 2.83 1.5E‐08 phenylalanine one 203 2.03 1.4E‐03 1 2.86 1.8E‐09 X‐17366 one 199 2.07 1.8E‐03 1 3.00 1.3E‐09 proline one 203 1.95 1.9E‐03 1 2.69 4.9E‐09 methionine one 203 2.04 1.9E‐03 1 2.90 1.4E‐09 lysine one 203 1.92 2.1E‐03 1 2.84 1.8E‐09 trigonelline (N'‐methylnicotinate) one 205 0.50 2.7E‐03 1 2.85 1.8E‐09 1,5‐anhydroglucitol (1,5‐AG) one 165 2.44 2.8E‐03 1 3.43 6.5E‐08 X‐18887 one 204 1.93 3.3E‐03 1 2.99 9.8E‐10 neopterin one 199 1.81 3.4E‐03 1 2.79 2.3E‐09 lactate one 201 1.88 3.9E‐03 1 2.89 2.3E‐09 X‐12690 one 204 1.90 4.4E‐03 1 3.00 6.6E‐10 isoleucine one 201 1.95 4.5E‐03 1 2.74 6.3E‐09 X‐18486 one 205 2.06 5.1E‐03 1 2.93 7.3E‐10 kynurenine one 203 1.87 5.4E‐03 1 2.86 3.4E‐09 3‐sialyllactose one 205 1.78 5.8E‐03 1 2.93 8.6E‐10 gentisate one 200 0.52 5.9E‐03 1 2.67 1.1E‐08 myo‐inositol one 205 1.94 6.0E‐03 1 2.94 1.6E‐09 5‐oxoproline one 204 1.86 6.8E‐03 1 3.03 6.4E‐10 xanthine one 205 0.54 7.1E‐03 1 2.79 4.4E‐09 kynurenate one 203 1.80 7.4E‐03 1 2.79 2.4E‐09 ornithine one 203 1.79 7.6E‐03 1 2.84 2.1E‐09 X‐12364 one 188 1.92 8.4E‐03 1 2.99 1.7E‐08 X‐12125 one 204 1.81 9.4E‐03 1 2.98 1.1E‐09 X‐12100 one 202 1.87 9.5E‐03 1 3.15 6.5E‐10 X‐12637 one 203 1.76 1.0E‐02 1 2.98 1.5E‐09 X‐12855 one 173 1.92 1.0E‐02 1 2.58 1.7E‐07 valine one 202 1.75 1.1E‐02 1 2.72 4.2E‐09 X‐13737 one 203 1.76 1.2E‐02 1 2.96 9.7E‐10 arginine one 190 1.78 1.2E‐02 1 2.75 5.5E‐09 4‐hydroxyphenylacetate one 205 0.57 1.2E‐02 1 2.69 5.0E‐09 pipecolate one 198 1.72 1.3E‐02 1 2.69 4.1E‐09 1‐(3‐aminopropyl)‐2‐pyrrolidone one 195 1.81 1.3E‐02 1 2.90 2.0E‐09 X‐12221 one 194 0.57 1.3E‐02 1 2.67 9.0E‐09 homovanillate (HVA) one 204 0.57 1.3E‐02 1 2.67 5.1E‐09 X‐13723 one 197 0.54 1.4E‐02 1 2.63 7.6E‐09 X‐12707 one 205 1.76 1.4E‐02 1 3.10 5.2E‐10 X‐12039 one 204 0.59 1.4E‐02 1 2.80 1.6E‐09 phosphate one 205 1.75 1.5E‐02 1 2.90 1.1E‐09 X‐16947 one 202 1.67 1.6E‐02 1 2.84 8.0E‐10 cysteine one 197 1.65 1.6E‐02 1 3.02 1.1E‐09 X‐18475 one 194 1.76 1.7E‐02 1 2.87 3.3E‐09 X‐14625 one 199 1.64 1.8E‐02 1 2.74 1.6E‐09 urate one 205 1.73 1.9E‐02 1 2.77 1.5E‐09 X‐12906 one 203 1.70 2.2E‐02 1 3.04 1.6E‐09 uracil one 205 0.61 2.2E‐02 1 2.62 3.2E‐09 X‐12685 one 186 0.56 2.2E‐02 1 2.64 1.1E‐08 vanillylmandelate (VMA) one 203 1.65 2.4E‐02 1 3.12 1.1E‐09 X‐12225 one 203 1.60 2.6E‐02 1 2.98 9.7E‐10 acetylcarnitine one 205 1.61 2.6E‐02 1 2.77 3.1E‐09 homocitrulline one 201 1.55 2.7E‐02 1 2.85 1.8E‐09 glutamine one 203 1.67 2.7E‐02 1 2.77 1.5E‐09

38 of 39 X‐18935 one 188 1.57 3.0E‐02 1 3.00 4.9E‐09 2‐hydroxybutyrate (AHB) one 194 1.64 3.0E‐02 1 3.27 2.0E‐09 X‐11945 one 202 1.63 3.1E‐02 1 2.90 8.3E‐10 hydantoin‐5‐propionic acid one 189 1.57 3.2E‐02 1 2.83 3.5E‐09 1,2‐propanediol one 204 0.63 3.3E‐02 1 2.97 8.6E‐10 scyllo‐inositol one 204 1.59 3.4E‐02 1 3.03 8.4E‐10 galactitol (dulcitol) one 192 1.58 3.4E‐02 1 2.78 3.3E‐09 X‐12379 one 168 1.62 3.6E‐02 1 2.54 9.2E‐08 X‐11423 one 205 1.55 3.6E‐02 1 2.91 6.1E‐10 1‐methylxanthine one 186 0.59 3.6E‐02 1 2.75 1.0E‐07 fucitol one 203 1.60 3.6E‐02 1 2.90 8.3E‐10 fructose one 205 1.58 3.8E‐02 1 2.88 8.6E‐10 alanine one 203 1.58 3.8E‐02 1 2.79 1.2E‐09 4‐imidazoleacetate one 174 0.58 3.8E‐02 1 2.77 1.5E‐07 azelate (nonanedioate) one 185 0.62 3.9E‐02 1 2.68 1.4E‐08 tryptophan one 205 1.59 3.9E‐02 1 2.84 9.8E‐10 inosine one 175 1.59 3.9E‐02 1 3.00 2.0E‐08 threonate one 205 1.57 3.9E‐02 1 2.93 6.8E‐10 X‐12230 one 176 0.60 4.0E‐02 1 2.46 7.4E‐07 sucrose one 204 1.56 4.0E‐02 1 2.87 5.3E‐10 4‐acetamidobutanoate one 204 1.52 4.1E‐02 1 2.89 7.2E‐10 X‐12860 one 189 1.57 4.3E‐02 1 2.87 1.2E‐08 X‐12708 one 203 1.50 4.5E‐02 1 2.95 9.9E‐10 xanthosine one 200 0.63 4.6E‐02 1 2.58 1.4E‐08 X‐15503 one 203 1.48 4.8E‐02 1 2.83 9.8E‐10 X‐12206 one 205 1.57 4.8E‐02 1 2.89 8.5E‐10 X‐13726 one 199 1.53 4.9E‐02 1 2.77 4.0E‐09 allantoin one 196 1.60 5.0E‐02 1 2.70 3.6E‐09

39 of 39