Livers were homogenized in ice-cold PBS using the OMNI Bead Ruptor and lipids extracted by mild acid methanolysis using concentrated HCl supplemented with trinonadecanoin (Nu-chek Prep, T-165) as internal standard for fatty acid methyl esters (FAMES) analysis. After extraction of resulting FAMES with 1 ml hexane, 20 μl of sample was analysed for FAMES by gas chromatography–mass spectrometry (GC–MS) using an Agilent 7890B/5977A with DB-WAX UI column (Agilent, 122–7032 UI). Complete GC–MS configurations and running programs are available upon request. Quantification of all ions was determined using custom python Tkinter software by comparison of sample data with serial dilutions of Nu-Check-Prep Fatty Acid Standard Mix (GLC 20a) that contain a mixture of various long-chain fatty acid species (methyl myristate, palmitate, palmitoleate, stearate, oleate, linoleate and linolenate). Integration of all ions (samples and standards) was performed on MassHunter Quantitative Analysis Program (Agilent Technologies). De novo lipogenesis analysis was evaluated by labelled isotopic enrichment of FAMES after correcting for the natural abundance of stable isotopes using the modern least-squares implementation of the skewed matrix correction method63 (link).
Masshunter quantitative analysis program
Manufactured by Agilent Technologies
The MassHunter Quantitative Analysis Program is a software application developed by Agilent Technologies for data processing and analysis of mass spectrometry data. The program is designed to facilitate the quantification of target compounds in complex samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using masshunter quantitative analysis program
1
Quantitative Liver Fatty Acid Profiling
2
Quantifying Cellular Fatty Acid Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were cultured in a 1:1 ratio of U13C glucose tracer for 24 h. Prior to collection, cells were imaged on Molecular Devices ImageXpress XL to assess cell numbers. Then cells were dissolved in 6 M guanidine HCl and transferred to glass tubes for derivitiziation with 3 M methanolic guanidine HCl. Samples were prepared alongside standard curve samples made up of FAMES mix (Nu-chek Prep, GLC 20a) and Cholesterol (Sigma, C8667).
Total cellular fatty acids were prepared by mild acid methanolysis66 (link) with the modifications. Integration and quantification was performed on MassHunter Quantitative Analysis Program (Agilent Technologies, B.06.00). Analysis for total quantification of fatty acids and Cholesterol and relative contributions of synthesis to the respective pool over labeling period were determined by fitting the isotopologue distributions to isotopomer spectral analysis (ISA).
Total cellular fatty acids were prepared by mild acid methanolysis66 (link) with the modifications. Integration and quantification was performed on MassHunter Quantitative Analysis Program (Agilent Technologies, B.06.00). Analysis for total quantification of fatty acids and Cholesterol and relative contributions of synthesis to the respective pool over labeling period were determined by fitting the isotopologue distributions to isotopomer spectral analysis (ISA).
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!