Coupling of Two-Dimensional Thin-Layer Chromatography with Gas Chromatography for the Quantitative Analysis of Lipid Classes and their Constituent Fatty Acids

The purpose of this study was to identify compounds in the wax of fruit obtained from different canopy positions for possible relationships to chilling injury development. Both exterior and interior canopy grapefruit wax extracts were separated by thin layer chromatography into four lipid classes: alkanes, aldehydes, linear alcohols and triterpenes. The triterpene class consisted of three subclasses: triterpenols (TOH), triterpenones (TON) and triterpenol acetates (TAC). Each triterpene subclass was analyzed by gas chromatography and their component structures estimated by comparison of retention times with authentic standards. Derivatives were prepared for each subclass: TOH were oxidized to TON, TON were reduced to TOH, and TAC hydrolyzed to TOH. Structures were verified by gas chromatography-mass spectra of components and their derivatives as well as spectra of authentic standards and their derivatives. There was a higher level of TON in interior compared with external canopy fruit. The three major TON were a amyrone, β amyrone and friedelin. α amyrone was also shown to be a major component of the wax of `Valencia' orange, a fruit not subject to chilling injury when produced in Florida

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Identification of Plasma Fatty Acids in Four Lipid Classes to Understand Energy Metabolism at Different Levels of Ketonemia in Dairy Cows Using Thin Layer Chromatography and Gas Chromatographic Techniques (TLC-GC)

Animals ◽

10.3390/ani10040571 ◽

2020 ◽

Vol 10 (4) ◽

pp. 571 ◽

Cited By ~ 1

Author(s):

Enrico Fiore ◽

Rossella Tessari ◽

Massimo Morgante ◽

Matteo Gianesella ◽

Tamara Badon ◽

...

Keyword(s):

Fatty Acids ◽

Thin Layer ◽

Dairy Cows ◽

Thin Layer Chromatography ◽

Metabolic Diseases ◽

Lipid Classes ◽

Plasma Fatty Acids ◽

Chromatographic Techniques ◽

Holstein Friesian ◽

Layer Chromatography

Excessive mobilization of adipose tissue in high milk producing dairy cows predisposes to metabolic diseases. The aim of this research was to identify the plasma fatty acids in four lipid classes as biomarkers for the diagnosis of hyperketonemia in bovines using thin layer chromatography and gas chromatographic techniques (TLC-GC). Sixty multiparous Holstein–Friesian dairy cows were enrolled in the study. Blood samples from the coccygeal vein were collected and β-hydroxybutyrate (BHB) was evaluated. All animals were divided into three groups on the basis of ketonemia: BHB < 0.50 mmol/L, 0.50 < BHB < 1.0 mmol/L, and BHB > 1.0 mmol/L. Plasma fatty acid concentrations were evaluated in four lipid classes: Free Fatty Acids (FFA), Triglycerides (TG), Cholesterol Esters (CE) And Phospholipids (PL). The concentration of fatty acids was analyzed using TLC-GC. The results showed the following significance in the lipid classes: 19 fatty acids were significant (p < 0.053) in FFA, nine fatty acids were significant (p < 0.050) in TG, eight fatty acids were significant (p < 0.050) in CE and three fatty acids were significant (p < 0.049) in PL. Eleven parameters were considered as predictive fatty acids related to animals in hyperketonemia. The FFA increased simultaneously with blood BHB levels, although the identified predictive fatty acids related to the TG and CE lipid classes decreased, meanwhile the BHB values increased. In the PL lipid class, no fatty acids were predictive.

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