Purification lipids

Goffic, F.L. (1997) Countercurrent and Centrifugal Partition Chromatography as New Tools for Preparative-Scale Lipid Purification, Lipid Technol. 9,148-150. 

Applications of countercurrent distribution to lipid purification were already reported in the 1950s. These included the isolation of PC, SPM, or cerebrosides from brain tissue, or the placenta. It was then mentioned that lipids easily emulsify, and this adversely affects the ability to separate them. Therefore these methods were only used for crude separation. The method also requires a long time for phase separation before each phase transfer, and this procedure needs to be repeated 500-3000 times. Otsuka and Yamakawa reported the application of droplet CCC to the purification of phospholipids and glycohpids. Because the stationary phase retention is much more stable in TC-CCC than with HS-CCC, it has become possible to select appropriate two-phase solvent systems. In this study, we showed the successful separation of human brain lipids by using TC-CCC. Additionally, if an isolated band can be observed on HPTLC, the lipid can be purified by using silica column chromatography after TC-CCC cmde separation. 

Yeasts have several difficulties for Upid extraction, including the presence of a thick cell wall that renders the yeast cells resistant to many solvents, as well as the possible presence of lipases in their cell extracts, and most of the neutral lipids are intracellularly stored in lipid bodies. However, lipid bodies also contain other lipophilic compounds, in particular aromatic compounds, which are difficult to remove during lipid purification (Ageitos et al. 2011). Table 7 shows the profile of TAG for some lipid extraction methods from L. starkeyi. 

The fatty adds commonly encountered in biological systems are straight chained alkanoic or alkenoic adds, containing an even number of carbon atoms (usually Ch-Ch). natural n Senera / these fatty adds can be produced readily by extraction of the lipids from sources natural sources and saponifying the neutral triglycerides. This is satisfactory providing a mixture of fatty acids is acceptable. Purification of spedfic fatty adds from the saponification mixture increases the costs considerably. 

Bligh, E.G. and E)yer, W.J. 1959 A rapid method of total lipid extraction and purification. Canadian Journal of Biochemical Physiology 37 911-917. 

As plant extracts mainly comprise large amonnts of ballast substances (e.g., lipids and chlorophylls), their purification is often a priority in the analysis. Such purification can be expensive in terms of both time and solvent consumed and can lead to losses of sample components. Online purification and separation of extracts contaminated with plant oil, can be readily performed by TLC in equilibrium chambers [1] that enable the use of continuous elution. 

Preliminary purification of a starting band contaminated with plant oil should be performed by predevelopment with a nonpolar solvent such as benzene or n-heptane, delivered from the eluent container. Weakly retained ballast substances (e.g., lipids) move with the solvent to the edge of the adsorbent layer, covering the glass plate where the volatile solvent evaporates. The contaminants can then be removed (scraped out with the adsorbent) from the layer or adsorbed on the strip of blotting paper placed on the upper edge of the layer. 

Tsuchida, K., Soulages, J. L., Moribayashi, A., Suzuki, K., Maekawa, H., and Wells, M. A. 1997. Purification and properties of a lipid transfer particle from Bombyx mori Comparison to the lipid transfer particle from Manduca sexta. Biochim. Biophys. Acta, 1337(l) 57-65. 

Heydenreich, A.V., Westmeier, R., Pedersen, N., Poulsen, H.S., and Kristensen, H.G., Preparation and purification of cationic solid lipid nanospheres effects on particle size, physical stability and cell toxicity, International Journal of Pharmaceutics, 2003, 254, 83-87. 

Liposomes containing PE lipid components may be activated with these crosslinkers to contain iodoacetyl derivatives on their surface (Figure 22.29). The reaction conditions described in Chapter 5, Section 1.5 may be used, substituting a liposome suspension for the initial protein being modified in that protocol. The derivatives are stable enough in aqueous solution to allow purification of the modified vesicles from excess reagent (by dialysis or gel filtration) without... 

By means of gel electrophoresis on cross-linked, hydrolyzed starch,99 with simultaneous checking for proteins, lipids, and pectinesterase activity, it was found, however, that the product isolated after the separation on CM-Sephadex C-50 constitutes but one of five multiple forms of tomato pectinesterase, and is the one present in preponderant proportion98 (see Fig. 4). The accompanying lipid and sugar components were separated from this pectinesterase form in the course of the purification procedure. After analysis of the hydro-lyzate of the final product for fatty acids, as well as for carbohydrate components, it was possible to exclude the possibility of a lipoprotein,30 as well as glycoprotein,100 character of this form of tomato pectinesterase. 

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