Summary of Study ST000472
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 PR000361. The data can be accessed directly via it's Project DOI: 10.21228/M8CW21 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000472 |
| Study Title | Effect of the culture medium on the degradation of nucleotide triphosphates |
| Study Type | Endpoint measurement |
| Study Summary | The effect of the Verduyn culture medium on the degradation profiles of nucleotide triphosphates extracted under typical boiling ethanol conditions was evaluated. |
| Institute | University of Groningen |
| Department | Analytical Biochemistry |
| Last Name | Bischoff |
| First Name | Rainer |
| Address | Antonius Deusinglaan 1 (XB20), Building 3226, room 601, 9713 AV Groningen, The Netherlands |
| r.p.h.bischoff@rug.nl | |
| Phone | NA |
| Submit Date | 2016-09-13 |
| Raw Data Available | Yes |
| Analysis Type Detail | LC-MS |
| Release Date | 2016-10-10 |
| Release Version | 1 |
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Sample Preparation:
| Sampleprep ID: | SP001178 |
| Sampleprep Summary: | Tubes containing 496 µL of 75% (aq) ethanol were preheated at 95°C for 5 min, followed by the addition of 4 µL of each nucleotide standard solution (500 µM) and vigorous mixing. 4 µL of solutions “A” to “H” were added to the reaction mixture and the samples were incubated at 95°C under shaking for 0 and 15 min. Reactions were stopped by snap-freezing in liquid nitrogen and samples were stored on dry ice. Subsequently, samples were thawed and 20 µL of the 13C15N-labeled internal standard solution were added for quantitative analysis. Excess solvent was evaporated under a stream of nitrogen without heating. Finally, samples were reconstituted in 200 µL of acetonitrile-water 70:30 and stored at –40°C until analysis by LC-MS. After the experimental procedure the final concentration of all components in solutions “A” to “H” was 10 µM. Compounds to be tested were divided into eight groups containing approximately ten compounds each, mainly comprising major central carbon metabolites including amino acids, organic acids, sugar phosphates, coenzymes, etc (Table 1). |
| Processing Method: | Preparation of reagents, dilution, solvent removal under a stream of nitrogen without heating and reconstitution |
| Extraction Method: | Boiling ethanol (95°C) |
| Extract Enrichment: | Evaporation of excess of solvent under a stream of nitrogen without heating. |
| Extract Storage: | -40C |
| Sample Resuspension: | 200 µL of acetonitrile-water 70:30 |
