Fatty Acid Oxidation (FAO) disorder are traditionally hypoketosis and hypoglycemia in newborns. Therefore, it is vital to carry out more specific diagnosis of such metabolic disorders due to the complications of biochemical processes in mitochondria. Such disorders are rare inherited that limit the - oxidation containing fatty acids through the citric cycle. We recently Identified the regulation of the xenometabolites derivatives cis- and trans-3,4-methylene heptanoyl carnitine in plasma and skeletal muscle of exercising humans.  We are presenting a developed method for the synthesis and extraction of medium, and long, odd chain cyclopropyl fatty acids CpFA that formed during gut microbe metabolism (plasma xenometabolites).  We modified the Folch method, and it was used for the extraction of lipids and more specifically cyclopropyl fatty acids from several biological matrixes such as adipose tissue, plasma, feces, and urine. Using the Foch method allows the purification and analysis of the free fatty acids in plasma tissue.  Following the extraction, further qualitative analysis of samples was done using gas chromatography mass spectroscopy GCMS. We developed a GC chromatogram with several authentic CpFA, and we used the new panel for the analysis of the extracted fatty acids samples from plasma. Our data identified the C15 as well as the C17 CpFA among other fatty acids present in the plasma extracts. Furthermore, extracted samples were run in duplicates as the “spiked” and un spiked of each plasma tissue and these fatty acids was identified, despite their chemical differences. Nonetheless, these results indicate that gas chromatography with mass spectroscopy techniques is a unique approach for the analysis of CpFA in plasma tissue samples.  Additionally, the synthesized CpFA will be purified using high performance liquid chromatography-HPLC and the spectroscopic characterization of the free cyclopropyl fatty acids derivatives will be achieved using 400 MHz NMR, GC/MS EI-QQQ, and LC/MS QTRAP 5500. The synthesized two odd-chain CpFA will be used as biomarkers in cell bioactivity assays.

Qualitative analysis and characterization of cyclopropyl fatty acids from plasma

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