Quantification of Acyl-CoA Metabolites

Acyl-CoA extraction was performed as previously described[20 (link)] after spiking in 100 μL of acyl-CoA internal standards derived from pan6 deficient Saccharomyces cerevisiae grown in [¹³C₃¹⁵N₁]-pantothenic acid containing media as previously described[21 (link)]. Since this acyl-CoA internal standard is biologically derived, some batch to batch variation is expected, but our previous calculations on yield estimate 100 μL of acyl-CoA internal standards produced by this method to contain 200 ng of [¹³C₃¹⁵N₁]-acetyl-CoA, 20 ng of [¹³C₃¹⁵N₁]-succinyl-CoA, and 5 ng of [¹³C₃¹⁵N₁]-propionyl-CoA as well as varying amounts of other [¹³C₃¹⁵N₁]-acyl-CoAs not included in this study.

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Frey A.J., Feldman D.R., Trefely S., Worth A.J., Basu S.S, & Snyder N.W. (2016). LC-quadrupole/Orbitrap high resolution mass spectrometry enables stable isotope resolved simultaneous quantification and 13C-isotopic labeling of acyl-Coenzyme A thioesters. Analytical and bioanalytical chemistry, 408(13), 3651-3658.

Publication 2016

Acetyl coa Acyl coas Pantothenic acid Propionyl coa Saccharomyces cerevisiae Succinyl coa

Corresponding Organization :

Other organizations : Drexel University, Bristol-Myers Squibb (United States), Brigham and Women's Hospital, Harvard University

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Variable analysis

independent variables

Spiking in 100 μL of acyl-CoA internal standards derived from pan6 deficient Saccharomyces cerevisiae grown in [13C3 15N1]-pantothenic acid containing media

dependent variables

Acyl-CoA levels, including 200 ng of [13C3 15N1]-acetyl-CoA, 20 ng of [13C3 15N1]-succinyl-CoA, and 5 ng of [13C3 15N1]-propionyl-CoA, as well as varying amounts of other [13C3 15N1]-acyl-CoAs

control variables

The acyl-CoA internal standard is biologically derived, so some batch to batch variation is expected

controls

The acyl-CoA internal standard derived from pan6 deficient Saccharomyces cerevisiae grown in [13C3 15N1]-pantothenic acid containing media serves as a positive control

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