Acyl CoAs Analysis Service

Acyl-CoAs are essential for many physiological processes, like lipid synthesis, fatty acid oxidation, ketone body generation, xenobiotic metabolism, and signaling transduction. Generally speaking, acyl-CoAs like free CoA, acetyl-CoA, and malonyl-CoA assist in regulation of metabolic flux. For example, glycolysis and fatty acid oxidation are regulated by the ratio of acetyl-CoA to free CoA. Another example is that the elevation of malonyl-CoA will lead to the synthesis of fatty acids and thereby inhibiting fatty acid oxidation by preventing acyl-CoA going into the mitochondrion. What’s more, the deacylation and acylation caused by various acyl-CoAs, like acetyl-CoA, palmitoyl-CoA and succinyl-CoA, plays an essential role in the function regulation of many proteins and genes. Acting as allosteric regulators of several enzymes and proteins, acyl-CoAs have a central role in many basic pathways.

However, there are various kinds of acyl-CoAs in the cell and may involve many other compounds besides fatty acids and their oxidized derivatives. Acyl-CoAs only exist inside the cells and their profiling inside the cells can give you some clues of the local metabolic status. Since each organ exerts its specific function and has its specific energetic demand, each organ is therefore expected to show a different acyl-CoA profile. For example, to meet the ATP demand for heart contraction, the heart tissue prefers utilizing fatty acids. However, in the brain, glucose and ketone bodies are primary substrate for energy generation. Meantime, in the liver, both the oxidation and synthesis of fatty acid are highly active. The profiles of the acyl-CoAs of these tissues broadly reveal the energy source preferences of these organs.

The need for a method to extract, separate, and quantify various amounts variety of acyl-CoAs with limited amounts of biological material increasing dramatically. Numerous methods have been developed to meet the requirement to measure acyl-CoAs in different biological tissues. Quantitative profiling of acyl-CoAs is a huge challenge for analytical world. This is because there are various kinds of acyl-CoAs and their concentrations in different tissues are in a wide range. What’s more, the wide range of polarity of the acyl-CoAs poses a challenge for LC method development. Though alkaline mobile phases can be used to improve the shapes of acyl-CoAs, their application is harmful to silica-based C18 columns and dramatically shortens the lifespan. Most methods, only analyze one kind of acyl-CoAs, short-, medium-, or long-chain. So far, MS is the most sensitive and reliable method for acyl-CoAs studies. Various LC-MS/MS approaches have been developed to enhance the sensitivity. Based on the fragmentation pattern, a MRM method allows the selection of both precursor and product ions and thereby ensure quantitatively profile of Acyl-CoAs with high sensitivity and accuracy.

Platform

• LC-MS/MS

Summary

• Identification and quantification of acyl-CoA species by mixed organic solvent extraction in cells or tissue. After extraction, acyl-CoA species are dried, re-suspended for LCMS separation and measured with LC-MS mass spectrometer using MRM methods.

Sample Requirement

• Normal Volume: 200ul serum/plasma; 200 mg tissue, 2e7 cells
• Minimal Volume: 50ul serum/plasma; 50 mg tissue, 5e6 cells

Report

• A detailed technical report will be provided at the end of the whole project, including the experiment procedure, MS/MS instrument parameters
• Analytes are reported as uM or ug/mg (tissue), and CV's are generally<10%
• The name of the analytes, abbreviation, formula, molecular weight and CAS# would also be included in the report.

With integrated set of separation, characterization, identification and quantification systems featured with excellent robustness & reproducibility, high and ultra-sensitivity, Creative Proteomics provides reliable, rapid and cost-effective Acyl CoAs targeted metabolomics services.