Approximately 30 mg of frozen tissues were submerged in liquid N2 and pulverized to powder in a SPEX, Inc. 6770 Freezer/Mill. Metabolites were extracted by solvent partitioning in acetonitrile:water:chloroform (2:1.5:1).(17 (link)) The upper polar metabolite fractions were frozen in several aliquots each and lyophilized overnight. One fraction of the resulting residue was reconstituted in 55 μL of D2O containing 25 nmol DSS-d6 and transferred to disposable 1.7 mm glass NMR tubes (Wilmad). NMR spectra were recorded at 14.1 T, 20° C, on an Agilent DD2 spectrometer, using standardized parameters as described previously.(18 (link)) Ion Chromatography-Mass Spectrometry (ICMS) was performed as previously described.(19 (link))
Protocol detail
Metabolite Extraction and NMR Analysis
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Markey Cancer Center, University of Kentucky, National Cancer Institute, National Institutes of Health, Center for Cancer Research
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Liquid nitrogen treatment
- Pulverization of frozen tissues in a SPEX, Inc. 6770 Freezer/Mill
- Metabolite extraction by solvent partitioning in acetonitrile:water:chloroform (2:1.5:1)
- Metabolite profiles analyzed by NMR spectroscopy
- Metabolite profiles analyzed by Ion Chromatography-Mass Spectrometry (ICMS)
- Tissue sample size (approximately 30 mg of frozen tissues)
- Temperature (20°C for NMR analysis)
- Magnetic field strength (14.1 T for NMR analysis)
- Use of DSS-d6 as an internal standard for NMR analysis
- No positive or negative controls were explicitly mentioned in the provided information.
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!