Summary of Study ST001628
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 PR001042. The data can be accessed directly via it's Project DOI: 10.21228/M80X2Z This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001628 |
| Study Title | Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Liver (part-V) |
| Study Type | Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR |
| Study Summary | This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis. |
| Institute | University of Florida |
| Department | Applied Physiology and Kinesiology |
| Laboratory | Rm 42 and Rm 43 |
| Last Name | Ryan |
| First Name | Terence |
| Address | 1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA |
| ryant@ufl.edu | |
| Phone | 352-294-1700 |
| Submit Date | 2020-12-11 |
| Num Groups | 2 |
| Total Subjects | 17 |
| Num Males | All |
| Study Comments | CKD metabolomic study via NMR using mice model |
| Publications | MDPI |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Analysis Type Detail | NMR |
| Release Date | 2021-06-30 |
| Release Version | 1 |
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Sample Preparation:
| Sampleprep ID: | SP001711 |
| Sampleprep Summary: | A modified form of FOLCH extraction protocol was used to extract metabolites from the tissues. Wet weights of all tissue samples were recorded prior to extraction. Tissue samples were immediately homogenized to prevent any possible enzymatic action using 1 mL of ice-cold methanol in a PowerLyzer 24 Homogenizer (QIAGEN Group, Hilden, Germany). The mixture was centrifuged using 13,200 rpm at 4oC for 30 minutes and the resulting supernatant was transferred to a new glass vial consisting 3 mL of ice cold chloroform:methanol (2:1, v/v) mixture. The homogenate was vortexed and left in an ice bath for 15 minutes to allow for phase separation. Next, 1 mL of 0.9% of saline was added, vortexed it for couple of minutes followed by a second incubation in an ice bath for 30-45 min for complete phase separation. The upper aqueous layer was transferred to a new falcon tube. To the remaining organic phase sample, 1 mL of 0.9% of saline was added again followed by vigorous mixing and letting it stand in ice bath (15 minutes) for a second phase separation. This second aqueous phase was combined with the first. The resulting aqueous and organic layers were dried separately. The aqueous layer was dried overnight with a Labconco freezer dryer (Labconco Corporation, MO, USA) and the organic layer was dried via inert nitrogen gas. These two dried powders (aqueous and organic phases) were stored at -80oC until performing NMR experiments. |
| Processing Method: | Lyophilization and Homogenization |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Modified FOLCH extraction |
| Extract Storage: | -80℃ |
| Sample Resuspension: | In 50 microliter of 50 mM phosphate buffer (pH 7.2) with 2 mM EDTA, 0.5 mM DSS and 0.2% sodium azide for aqueous phase samples. |
| Sample Spiking: | 0.5 mM of DSS for aqueous phase samples |
