Fluorescence fatty acids

W. E. Harris and W. L. Stahl, Incorporation of cis-parinaric acid, a fluorescent fatty acid, into synaptosomal phospholipids by an acyl-CoA acyltransferase, Biochim. Biophys. Acta 736, 79-91 (1983). 

Glutathione S transferases bind bile acids in vitro but doubt has been cast over whether this happens in vivo as these enzymes were not labelled by fluorescently labelled bile acids in experiments to identify the carrier proteins but may play a role with the raised levels in cholestasis. Liver fatty-acid-binding protein has been shown to bind bile acids by using a displacement assay with fluorescent fatty-acid ligand. This work clearly showed displacement to be directly related to hydrophobicity, such that lithocholate conjugates had the greatest effect. This may indicate a mechanism to minimise toxicity within the hepatocyte. 

Phosphatidylinositol containing fluorescent fatty acids has also been prepared [303] by synthesising a phosphatidylcholine containing fluorescent fatty acids and then using a phospholipase D, in the presence of inositol, to effect an ester interchange. 

Fluorescent fatty acids using fatty acid trafficking and signaling... 

Kleine, and S. Petry, Analysis of lipolysis in adipocytes using a fluorescent fatty acid derivative, Biochirnie, in press. 

Doody, M. C., Pownall, H. J., Kao, Y. J. and Smith, L. C., Mechanism and kinetics of transfer of a fluorescent fatty acid between single-walled phosphatidylcholine vesicles, Biochemistry, 19, 108-116 (1980). 

Fibach, E. Giloh, H. Rachmilewitz, E. A. Gatt, S. Flow cytofluorometric analysis of the uptake of the fluorescent fatty acid pyrenedodecanoic acid by human peripheral blood cells. Cytometry 1988, 9, 525-528. 

Nahas, N. Fibach, E. Giloh, H. Gatt, S. Use of the fluorescence activated cell sorter for studying uptake of fluorescent fatty acids into cultured cells. Biochim. Biophys. Acta, Lipids LipidMetab. 1987,977,86-91. 

Browning Reactions. The fluorescent components formed in the browning reaction (8) of peroxidized phosphatidylethanolamine are produced mainly by interaction of the amine group of PE and saturated aldehydes produced through the decomposition of fatty acid hydroperoxides. 

Acids can also be converted to fluorescent dansyl derivatives The reaction of Cg to C24 fatty acids with dansyl semipiperazide or semicadavende provides an excellent example (Fig 34) [87] Odd-numbered and unsaturated fatty acids [88] and propionic, sorbic and benzoic acid [89] can be detected in the same manner... 

Palmitic and stearic acids aflatoxins Bi and Bi 15 to 35% unseparated fatty acids intensified the fluorescence when investigating com extracts [224]... 

Fig. 1 Fluorescence scan of a fatty acid mixture with 500 ng substance per chromatogram zone. Arachidic acid (I), stearic acid (2), palmitic acid (3), myristic acid (4), lauric acid (5).

Fatty Acid Transporters. Figure 2 Quencher-based real-time fatty acid uptake assay with a fluorescently labeled FFA analogue (C1-Bodipy-C12). Predominantly protein-mediated fatty acid uptake by 3T3-L1 adipocytes (diamonds) was compared with diffusion-driven uptake by fibroblasts (squares) using the QBT Fatty Acid Uptake reagent (Molecular Devices Corp., CA, USA), which contains C1-Bodipy-C12 as substrate in conjunction with a cell impermeable quencher. Uptake kinetics was recorded using a Gemini fluorescence plate reader. Error bars indicate the standard deviations from 12 independent wells. RFU relative fluorescence units. 

In vitro and ex vivo studies have shown that FATPs transport LCFAs and very long-chain fatty acids (VLCFAs) but no medium-chain fatty acids, fatty acid esters, or lipid-soluble vitamins [4]. LCFA transport is inhibited by prior protease treatment. Synthetic substrates for FATPs include 14C-labeled fatty acids and the fluorescently labeled fatty acid analogue C1 -BODEP Y-Cl 2. Using the latter substrate, differences in fatty acid uptake kinetics between FATP expressing 3T3 LI adipocytes and 3T3 LI fibroblasts, which are devoid of FATPs, can be readily appreciated (Fig. 2). 

Hydrazinocarbonylpropyl)-6,7-dimethoxy-l-methyl-2(177)-quinoxalinone (126) undergoes acylation by fatty acids (EtN=C=NCH2CH2CH2NMe2, H2O, pyridine, 20°C) to afford highly fluorescent derivatives that are useful for the detection and estimation of such fatty acids. ... 

The red microalga genus Porphyridium is a source of biochemicals possessing nutritional and therapeutic values. The biochemicals include polysaccharides that have anti-inflammatory and antiviral properties, long-chain polyunsaturated fatty acids, carotenoids such as zeaxanthin, and fluorescent phycobiliproteins. 

Chloroform-methanol extracts of Borrelia burgdorferi were used for the identification of lipids and other related components that could help in the diagnosis of Lyme disease [58]. The provitamin D fraction of skin lipids of rats was purified by PTLC and further analyzed by UV, HPLC, GLC, and GC-MS. MS results indicated that this fraction contained a small amount of cholesterol, lathosterol, and two other unknown sterols in addition to 7-dehydrocholesterol [12]. Two fluorescent lipids extracted from bovine brain white matter were isolated by two-step PTLC using silica gel G plates [59]. PTLC has been used for the separation of sterols, free fatty acids, triacylglycerols, and sterol esters in lipids extracted from the pathogenic fungus Fusarium culmorum [60]. 

FIG. 11 Order parameter variation along acyl chains in red cell ghosts ( ), small unilamellar vesicles of egg phosphatidylcholine (V), and paraffin oil (+), as determined by the fluorescence anisotropy decay of the w-anthroyloxy fatty acid probes. (Reprinted by permission from Ref. 12.)... 

It is of interest to examine the development of the analytical toolbox for rubber deformulation over the last two decades and the role of emerging technologies (Table 2.9). Bayer technology (1981) for the qualitative and quantitative analysis of rubbers and elastomers consisted of a multitechnique approach comprising extraction (Soxhlet, DIN 53 553), wet chemistry (colour reactions, photometry), electrochemistry (polarography, conductometry), various forms of chromatography (PC, GC, off-line PyGC, TLC), spectroscopy (UV, IR, off-line PylR), and microscopy (OM, SEM, TEM, fluorescence) [10]. Reported applications concerned the identification of plasticisers, fatty acids, stabilisers, antioxidants, vulcanisation accelerators, free/total/bound sulfur, minerals and CB. Monsanto (1983) used direct-probe MS for in situ quantitative analysis of additives and rubber and made use of 31P NMR [69]. 

Thin-layer chromatography is employed in many areas of QC and routine monitoring of product quality [458]. Fluorescence scanning, densitometry or videodensitometry are used for quantification. Not all polymer additives are amenable to TLC analysis. Some fatty acid amides are virtually insoluble in organic solvents and cannot be isolated by thin-layer or column chromatography. 

HPLC has also been used for analysing fatty acid mixtures [708] and for the characterisation of fatty acids and their derivatives [709]. Fatty acids are commonly analysed on polymeric RPLC columns. Only multiple unsaturated fatty acids can be detected by UV in HPLC the others require derivatisation into UV-absorbing or fluorescing derivatives. Simultaneous determination of saturated and unsaturated fatty acids (C12-C24) by means of RPLC has been reported [710]. Derivatisation is necessary. 

The photobleaching of P-carotene by fluorescent light in fatty acid ester solutions showed an autoxidation kinetic profile with the rate of degradation of P-carotene in the order laurate > oleate > linoleate (Carnevale et al. 1979). The presence of a radical scavenger retarded the autoxidation, thus leading to the view that protection against autoxidation is built into the system by the unsaturation in the fatty acid. 

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