scholarly journals 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 ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 571 ◽  
Author(s):  
Enrico Fiore ◽  
Rossella Tessari ◽  
Massimo Morgante ◽  
Matteo Gianesella ◽  
Tamara Badon ◽  
...  

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.

1975 ◽  
Vol 53 (11) ◽  
pp. 1170-1183 ◽  
Author(s):  
W. C. Breckenridge ◽  
A. Kuksis

The molecular specificity in the biosynthesis of diacylglycerols by rat intestinal mucosa was examined by means of radioactive markers, thin-layer chromatography with silver nitrate and gas-liquid chromatography with radioactivity monitoring. Bile salt micelles of alternately labeled monoacylglycerols and free fatty acids were incubated with everted sacs of intestinal mucosa for various periods of time and the diacylglycerols were isolated by solvent extraction and thin-layer chromatography. Stereospecific analyses of the X-1,2-diacylglycerols labeled from 2-monoacylgiycerols showed that the sn-1,2-isomers (45–55%) were slightly in excess of the sn-2,3-isomers (34–45%) with the X-1,3-diacylglycerols accounting for the rest of the radioactivity (5–10%). This suggests that racemic diacylglycerols may be intermediates in the resynthesis of dietary fat in rat intestinal mucosa. Detailed analyses of the molecular species of the sn-1,2-diacylglycerols labeled from free fatty acids revealed that 10–45% of the total did not contain the acid present in the 2-monoacylglycerol supplied, and therefore had originated from the phosphatidic acid pathway. These findings are at variance with those obtained in isolated microsomes, which have suggested an inhibition of the phosphatidic acid pathway by monoacylglycerols as well as have given evidence of an exclusive synthesis of sn-1,2-diacylglycerols from 2-monoacylglycerols.


Author(s):  
Dharmendra B. Sharma ◽  
Parth Aphale ◽  
Vineet Sinnarkar ◽  
Sohan S. Chitlange ◽  
Asha Thomas

Background: Chromatography is one of the important laboratory technique in which the components of a mixture are separated on an adsorbent in order to analyze, identify, purify and quantify a mixture. Thin Layer Chromatography (TLC)is used to support the identity of a compound in a mixture when the Rf of a compound is compared with the Rf of a known compound. High Performance Thin Layer Chromatography is a sophisticated and automated form of Thin Layer Chromatography (TLC). The procedure simultaneously processes the sample and standard that results in better analytical precision and accuracy at a faster pace. Pharmacological/ Toxicological action of Nux Vomica is because of its active principles present in the seeds namely strychnine, brucine etc. This research paper aims to corelate the active principles present in Nux Vomica with the toxicological action of the same. Materials and Methods: 1. Standard Nux Vomica mother tincture was tested for its alkaloid markers and its correlation with the toxicological action was studied. 2. Analysis of the mother tincture was done using High Performance Thin Layer Chromatography. 3. Stationary phase consisted of TLC Aluminium sheets with silica gel 60 F253 pre-coated layer (20cm x 10cm), thickness-0.2mm, no. of tracks-18, band length-6mm. 4. Mobile Phase consisted of Chloroform: Methanol (9.5:0.5). 5. The plate was developed in developing chamber and observed under U.V. Light. Results: Colours seen on the HPTLC Plates of samples are greenwhich corresponds to strychnine, dark blue which corresponds to brucine, orange to alkaloids fluorescent green to sterols and pink to fatty acids which are evident on the chromatogram. Conclusion: Therapeutic action of Nux Vomica as noted in Homoeopathic Materia Medica is because of the active principles like strychnine, brucine, alkaloids, sterols, fatty acids present in it which is evident from the chromatogram.


2020 ◽  
Vol 21 (5) ◽  
pp. 1670
Author(s):  
Akhikun Nahar ◽  
Anthony L. Baker ◽  
David S. Nichols ◽  
John P. Bowman ◽  
Margaret L. Britz

In addition to cell membrane phospholipids, Actinobacteria in the order Corynebacteriales possess a waxy cell envelope containing mycolic acids (MA). In optimized culture condition, some species can also accumulate high concentrations of intracellular triacylglycerols (TAG), which are a potential source of biodiesel. Bacterial lipid classes and composition alter in response to environmental stresses, including nutrient availability, thus understanding carbon flow into different lipid classes is important when optimizing TAG synthesis. Quantitative and qualitative analysis of lipid classes normally requires combinations of different extraction, derivatization, chromatographic and detection methods. In this study, a single-step thin-layer chromatography-flame ionization detection (TLC-FID) technique was applied to quantify lipid classes in six sub-Antarctic Corynebacteriales strains identified as Rhodococcus and Williamsia species. A hexane:diethyl-ether:acetic acid solvent system separated the total cellular lipids extracted from cells lysed by bead beating, which released more bound and unbound MA than sonication. Typical profiles included a major broad non-polar lipid peak, TAG and phospholipids, although trehalose dimycolates, when present, co-eluted with phospholipids. Ultra-performance liquid chromatography-tandem mass-spectrometry and nuclear magnetic resonance spectroscopy detected MA signatures in the non-polar lipid peak and indicated that these lipids were likely bound, at least in part, to sugars from cell wall arabinogalactan. Waxy esters were not detected. The single-solvent TLC-FID procedure provides a useful platform for the quantitation and preliminary screening of cellular lipid classes when testing the impacts of growth conditions on TAG synthesis.


Biomolecules ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1363 ◽  
Author(s):  
René Herrera ◽  
Jarl Hemming ◽  
Annika Smeds ◽  
Oihana Gordobil ◽  
Stefan Willför ◽  
...  

Hazelnut (HS) and walnut (WS) shells, an abundant by-product of the processing industries of these edible nuts, are traditionally considered as a low-value waste. However, they are a source of valuable compounds with an interesting chemical profile for the chemical and pharmaceutical sectors. In this study, the lipophilic and hydrophilic extracts present in HS and WS were quantified and identified, then the polar fractions were chromatographically separated, and their antioxidant capacity was studied. The experimental work includes the isolation of crude lipophilic and hydrophilic extracts by an accelerated extraction process, chromatographic analysis (gas chromatography-flame ionization (GC-FID), GC-mass spectroscopy (GC-MS), high-performance size-exclusion chromatography (HPSEC), thin-layer chromatography (TLC)), and quantification of the components. In addition, a thorough compositional characterization of the subgroups obtained by flash chromatography and their antioxidant capacity was carried out. The gravimetric concentrations showed different lipophilic/hydrophilic ratios (0.70 for HS and 0.23 for WS), indicating a higher proportion of polar compounds in WS than in HS. Moreover, the lipophilic extracts were principally composed of short-chain fatty acids (stearic, palmitic, and oleic acid), triglycerides, and sterols. The polar fractions were screened by thin-layer chromatography and then separated by flash chromatography, obtaining fractions free of fatty acids and sugar derivatives (97:3 in HS and 95:5 in WS), and mixtures richer in phenolic compounds and flavonoids such as guaiacyl derivatives, quercetin, pinobanksin, and catechin. The most polar fractions presented a higher antioxidant capacity than that of the crude extracts.


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