Lipids saponification

Alkali is a frequent cause of eye bums as again confirmed in a recent study of Midelfarth [22], Alkali reacts with the tissue surface by concentration and time-dependent dissolution of the Upid membranes of epithelial cells the chemical mechanism is saponification of lipids with loss of all membranous barriers. Lipid saponification of membranous lipids starts at a pH over 11 [23]. 

A lipid lowering agent of potential value in hypercholesterolemia is cetaben (31). It is synthesized facilely by monoalkylation of ethyl -aminobenzoate with hexadecyl bromide and then saponification. " ... 

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. 

Alkaline hydrolysis (saponification) has been used to remove contaminating lipids from fat-rich samples (e.g., pahn oil) and hydrolyze chlorophyll (e.g., green vegetables) and carotenoid esters (e.g., fruits). Xanthophylls, both free and with different degrees of esterification with a mixture of different fatty acids, are typically found in fruits, and saponification allows easier chromatographic separation, identification, and quantification. For this reason, most methods for quantitative carotenoid analysis include a saponification step. 

Saponification (see Section 7.4) is carried out to extract more recalcitrant lipids, and the yields are higher than for conventional solvent extraction (Stern et al. 2000). 3 ml of 0.5 M methanolic NaOH is added to 0.1 g of the shard powder and heated at 70°C for 3 hours in a sealed glass vial. After cooling, the supernatant is acidified with HC1 and extracted with three aliquots of 3 ml //-hexane. The hexane will not mix with the methanolic solution (unlike the DCM MeOH used above), but will absorb the lipids and can be transferred into a new clean vial. The removal of excess hexane is carried out as above. Saponification will hydrolyze and methylate any ester functionalities, which removes the requirement to derivatize the samples (Section 7.4) unless other molecules are suspected of being present. However, any wax esters or triacylglycerols will also be hydrolyzed to their fatty acid methyl esters and alcohols therefore, if information on their composition is required, then conventional solvent extraction is recommended as a first step. For subsequent characterization of the lipid extract, see Chapter 7. 

Mono-, di- and triacylglycerols may all be measured by determination of the amount of glycerol released by hydrolysis. The lipid is first extracted into chloroform-methanol (2 1) and saponification is performed under conditions that will not affect any phosphate ester bonds, otherwise glycerol originating from phosphoglycerides would also be measured. Heating at 70°C for 30 min with alcoholic potassium hydroxide (0.5 mol l-1) has been shown to be satisfactory. However, the phospholipids may be removed prior to saponification either by extraction or by adsorption on activated silicic acid. 

An alternative procedure involves the release of the fatty acids by alkaline hydrolysis (saponification) by refluxing the extracted sample with dilute alcoholic potassium hydroxide for 1 h. After cooling, adding water and acidifying, the fatty acids are extracted into diethyl ether. The methyl esters can then be prepared by treatment with diazomethane, which may also be used directly on free fatty acids. Saponification is less satisfactory, because it is a lengthy procedure and often results in the loss of lipid components. 

Few analytical methods are available for the determination of total petroleum hydrocarbons in biological samples, but analytical methods for several important hydrocarbon components of total petroleum hydrocarbons may be modified. Most involve solvent extraction and saponification of lipids, followed by separation into aliphatic and aromatic fractions on adsorption columns. Hydrocarbon groups or target compounds are determined by gas chromatography-flame ionization or... 

Retinoids The challenge in fat-soluble vitamins analysis is to separate them from the lipid fraction that contains interferents. Alkaline hydrolysis, followed by LLE, is widely applied to remove triglycerides. This technique converts the vitamin A ester to all-trani-retinol. A milder process, which does not hydrolyze vitamin A ester, is alcoholysis carried out with metha-nolic KOH solution under specific conditions that favor alcoholysis rather than saponification. A more accurate explanation of this technique is reported in the book Food Analysis by FIPLC [409]. For some kind of matrices a simple liquid extraction can be sufficient with [421-423] or without [424,425] the purification... 

Lipids, relatively nonpolar chemical substances found in plant, bacterial, and animal cells, are among the most ubiquitous of biomolecules. In this experiment, a lipid extract of ground nutmeg will be purified by chromatography on a silica gel column. Analysis of the lipid extract by thin-layer chromatography will provide the classification of the components in the extract. The unknown lipids will be further characterized by saponification and analysis of the fatty acid content by gas chromatography. For an abbreviated experiment, students may be provided samples of natural oils and fats that can be analyzed by saponification and gas chromatography. 

In the second part of this experiment you will characterize the purified lipids (triacylgiycerols) isolated from nutmeg. The fatty acids in the triacyl-glycerols are released by saponification and their identities determined by gas chromatography. Alternatively, students may be provided various fat and oil samples for analysis. For example, the fatty acid content of triacyl-... 

There are basically two mechanisms to convert the fatty acids in a complex lipid to fatty acid methyl esters (FAMEs) methylation following hydrolysis of the fatty acids from the complex lipids, or direct transesterification. The first mechanism involves saponification (alkaline hydrolysis) in which the ester bond is cleaved between the fatty acid and the glycerol moiety (e.g., triacylglycerols and phospholipids) under heat and in the presence of an alkali (usually sodium hydroxide), followed by methylation performed in the presence of an acidic catalyst in methanol. Direct transesterification is usually a one-step reaction involving alkaline or acidic catalysts. 

BASIC PROTOCOL I PREPARATION OF FATTY ACID METHYL ESTERS FROM LIPID SAMPLES CATALYZED WITH BORON TRIFLUORIDE IN METHANOL In this method, lipid samples are first saponified with an excess of NaOH in methanol. Liberated fatty acids are then methylated in the presence of BF3 in methanol. The resulting fatty acid methyl esters (FAMEs) are extracted with an organic solvent (isooctane or hexane), and then sealed in GC sample vials for analysis. Because of the acidic condition and high temperature (100°C) used in the process, isomerization will occur to those fatty acids containing conjugated dienes, such as in dairy and ruminant meat products, that contain conjugated linoleic acids (CLA). If CLA isomers are of interest in the analysis, Basic Protocol 2 or the Alternate Protocol should be used instead. Based on experience, this method underestimates the amount of the naturally occurring cis-9, trans-11 CLA isomer by -10%. The formulas for the chemical reactions involved in this protocol are outlined in Equation D1.2.1 Saponification RCOO-R + NaOH, RCOO-Na + R -OH v 100°C DC Esterification RCOO-Na + CH,OH r 3 v RCOO-CH, + NaOH ioo°c ... 

This unit defines three different tests that are used to evaluate lipid systems. The first two, i.e., iodine value (IV see Basic Protocol I) and saponification value (SV see Basic Protocol 2), are used to determine the level of unsaturation and the relative size (chain length) of the fatty acids in the system, respectively. The free fatty acid (FFA) analysis (see Basic Protocol 3) is self-explanatory. Each of these analyses provides a specific set of information about the lipid system. The IV and SV provide relative information this means that the data obtained are compared to the same data from other, defined lipid systems. In mixed triacylglyceride systems there is no absolute IV that indicates the exact number of double bonds or SV that indicates the exact chain length. The data from the FFA analysis is an absolute value however, the meaning of the value is not absolute. As a quality indicator, ranges of FFA content are used and the amount that can be tolerated is product and/or process dependent. 

As indicated previously, this analysis provides a measure of the mean molecular weight of the fatty acids in a lipid system. It is based on the fact that saponification breaks ester bonds in a lipid system and since fatty acids are attached to the glycerol backbone with an ester bond, SV reflects the number of ester bonds per gram sample. The saponification value indicates the mean molecular weight of the sample s triacylglycerols and when divided by 3 gives the mean molecular weight of the constituent fatty acids. Therefore, the smaller the... 

Again, as with the IV, the saponification value provides some basic information about a lipid system. In a pure system (Table DI.4.3), with a series of triglycerides of increasing chain length, as the chain length increases there is a decrease in saponification value. With triglycerides of the same or close chain lengths,... 

Saponification. Before solvent is added for extraction, saponification (alkaline hydrolysis) is a step used in most extractions of tocopherols and tocotrienols. It should be noted that acetate forms of tocopherols or tocotrienols in a sample are changed to free tocopherols and tocotrienols after saponification. This process breaks down the ester bonds of lipids and sample matrices as well. In most extraction procedures, a 60% to 80% (w/v) aqueous solution of KOH is used to perform the saponification. The volume of KOH required varies according to the amount of lipid contained in the sample. Also, ethanol is needed to stabilize the saponified solution and prevent the precipitation of soap material. Usually, the ratio of KOH, ethanol, and fat (in sample) during saponification is 3 (g) 15 (ml) 1 (g), respectively (Ball, 1988). The ratio may need to be adjusted based on the types of fats in the sample. Although ethanol concentration has no effect on the extraction of a-tocopherol by hexane, a concentration above 30% may cause lower recoveries of other tocopherols (Ueda and Igarashi, 1990). For most food samples, saponification for 30 min at70°C is sufficient. 

Lipid extraction prior to saponification is used in some food samples with high concentrations of proteins and carbohydrates. Some of these components are readily hydrolyzed by alkaline solution during saponification, which may generate various hydrolysis products that could interfere with solvent extraction and chromatography. Infant formula is a good example of a case where lipid extraction is performed before saponification. Tuan et al. (1989) indicated that... 

Sample preparation, protein analysis, 78 Saponification of lipids, 438-439... 

Following extraction, an efficient way of initiating the isolation of carotenoids is to saponify the extract. This removes many of the unwanted lipids present in the sample as well as chlorophyll. The saponification by-products, which to a great extent are sodium or potassium salts, are easily separated by an aqueous solution of a highly polar salt. The addition of water also helps wash off excess alkali and other water-soluble and water-complexed compounds. This procedure hydrolyzes xantho-phyll esters to form the hydroxylated carotenoid. 

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