Summary of Study ST002977
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 PR001853. The data can be accessed directly via it's Project DOI: 10.21228/M86B06 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 | ST002977 |
| Study Title | Offline Two-dimensional Liquid Chromatography-Mass Spectrometry for Deep Annotation of the Fecal Metabolome following Fecal Microbiota Transplant |
| Study Summary | In this study, we describe a novel experimental strategy using multidimensional chromatography to facilitate compound identification in untargeted metabolomics. Pooled fecal metabolite extract samples were fractionated using an offline semi-preparative liquid chromatography. The resulting fractions were analyzed by an orthogonal LC-MS/MS method, and the data were searched against commercial, public and local spectral libraries. Multidimensional chromatography yielded more than a 3-fold improvement in identified compounds compared to the typical single-dimensional LC-MS/MS approach, and successfully identified several rare and novel compounds including atypical conjugated bile acid species. Most features identified by the new approach could be matched to features that were detectable, but not identifiable, in the original single-dimensional data. An evaluation of this approach in the context of patients with recurrent Clostridioides difficile infection receiving fecal microbiota transplants is also included. Overall, our approach represents a powerful strategy for deeper annotation of the metabolome that can be implemented with common commercially-available instrumentation, and should be applicable to any dataset requiring deeper annotation of the metabolome. |
| Institute | University of Michigan |
| Department | Michigan Compound Identification Development Core |
| Last Name | Anderson |
| First Name | Brady |
| Address | 1000 Wall St, Ann Arbor, MI 48105 |
| anderbra@umich.edu | |
| Phone | 734-232-8177 |
| Submit Date | 2023-06-02 |
| Num Groups | 2 |
| Total Subjects | 8 |
| Publications | Publication to come later |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-03-11 |
| Release Version | 1 |
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Sample Preparation:
| Sampleprep ID: | SP003096 |
| Sampleprep Summary: | Fecal samples were weighed into pre-tared 2 mL Precellys (Bertin Corp.) compatible vials, and one 2.8 mm stainless steel bead was added to aid homogenization. Fecal matter was homogenized using a Precellys Evolution by two 20 s cycles separated by a 30 s break. Extraction solvent was used at a ratio of 1 mL per 5 g feces and was comprised of 1:1:1 methanol:acetonitrile:acetone containing 10 uM of D3-creatine, D10-isoleucine, D2-biotin, D5-tryptophan, D3-caffeine, D3-octanoylcarnitine, D3-palmitoylcarnitine, D4-deoxycholic acid, D4-cholic acid, and D7-arginine as internal standards. Following extraction, samples were centrifuged for 10 min at 17,000 rpm. 100 uL aliquots of supernatant were transferred to Eppendorf vials, dried under a gentle stream of nitrogen, and stored at -80 ºC. On the day of analysis, the dried extracts were reconstituted in 85:15 acetonitrile:water for HILIC analysis or 9:1 water:methanol for RPLC analysis with volumes as described below. Pooled samples were prepared by combining equal volumes of reconstituted fecal matter extracts from all subjects. |
| Processing Storage Conditions: | On ice |
| Extraction Method: | 1 g feces / 5 mL of 1:1:1 methanol:acetonitirle:acetone |
| Extract Storage: | 4℃ |
| Sample Resuspension: | 9:1 Water:Methanol for Reversed phase - CSH; 85:15 acetonitrile:water for HILIC |
