Summary of Study ST001192

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000804. The data can be accessed directly via it's Project DOI: 10.21228/M8RM32 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST001192
Study Title	A library of human gut bacterial isolates paired with longitudinal multiomics data enables mechanistic microbiome research
Study Type	Stool metabolite profiling
Study Summary	Fecal microbiota transplantation (FMT) is used in the treatment of microbiome-associated diseases such as Clostridium difficile infections. In order to develop synthetic therapeutics and customized disease treatments we will need to understand the bacterial communities in the stool samples used in such treatments. For this purpose, a microbiome library was generated using human stool obtained from healthy human FMT recruited by OpenBiome, a non-profit organization that provides fecal microbiome therapeutics. In addition to characterizing the bacterial populations and obtaining bacterial isolates from FMT samples, we conducted metabolite profiling with the goal of: (1) generating a library of metabolites in FMT samples, (2) Identifying metabolites associated with defined bacterial populations, and (3) identifying microbial metabolites with immunoregulatory functions. We conducted metabolite profiling on a subset consisting of 180 stool samples from 84 donors using four nontargeted liquid chromatography mass spectrometry (LC-MS) methods. Generated data were processed, isotopes removed, and adducts and fragments clustered. The identity of known metabolites was determined based on matching retention times of neat standards run in parallel with the study.
Institute	Broad Institute of MIT and Harvard
Last Name	Avila-Pacheco
First Name	Julian
Address	415 Main Street
Email	jravilap@broadinstitute.org
Phone	617-714-8264
Submit Date	2019-06-10
Total Subjects	84
Raw Data Available	Yes
Analysis Type Detail	LC-MS
Release Date	2019-07-17
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001267
Sampleprep Summary:	Raw stool were diluted 1:10 in 12.5% glycerol buffer and 0.9% NaCl, homogenized and filtered through a 330um filter. HILIC-pos: LC-MS samples were prepared from homogenate(10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. HILIC-neg: Stool homogenates (30μL) were extracted using 120 μL of 80% methanol (VWR) containing 0.05 ng/μL inosine-15N4, 0.05 ng/μL thymine-d4, and 0.1 ng/μL glycocholate-d4 as internal standards (Cambridge Isotope Laboratories). The samples were centrifuged (10 min, 9,000 x g, 4ºC) and the supernatants (10 μL) were injected directly. C18-neg: Stool homogenates (30 μL) were extracted using 90 μL of methanol containing PGE2-d4 as an internal standard (Cayman Chemical Co.; Ann Arbor, MI) and centrifuged (10 min, 9,000 x g, 4°C). C8-pos: Lipids were extracted from homogenates (10 μL) using 190 μL of isopropanol containing 1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard (Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000 x g, ambient temperature), supernatants (10 μL) were injected directly

Sampleprep ID:

SP001267

Sampleprep Summary:

Raw stool were diluted 1:10 in 12.5% glycerol buffer and 0.9% NaCl, homogenized and filtered through a 330um filter. HILIC-pos: LC-MS samples were prepared from homogenate(10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. HILIC-neg: Stool homogenates (30μL) were extracted using 120 μL of 80% methanol (VWR) containing 0.05 ng/μL inosine-15N4, 0.05 ng/μL thymine-d4, and 0.1 ng/μL glycocholate-d4 as internal standards (Cambridge Isotope Laboratories). The samples were centrifuged (10 min, 9,000 x g, 4ºC) and the supernatants (10 μL) were injected directly. C18-neg: Stool homogenates (30 μL) were extracted using 90 μL of methanol containing PGE2-d4 as an internal standard (Cayman Chemical Co.; Ann Arbor, MI) and centrifuged (10 min, 9,000 x g, 4°C). C8-pos: Lipids were extracted from homogenates (10 μL) using 190 μL of isopropanol containing 1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard (Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000 x g, ambient temperature), supernatants (10 μL) were injected directly