Preparation of FAME from sterol esters

Ichihara, Kazuo; Yamaguchi, Chiaki; Nishijima, Hiroto; Saito, Kazumi

doi:10.1007/s11746-003-0781-2

Cited by 8 publications

(5 citation statements)

References 5 publications

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“…As CEs are less reactive than TGs [22,23], it is reasonable to expect that no FAMEs would be produced from CEs with KOH in aqueous methanol. The accuracy and usefulness of selective methanolysis were confirmed with seed oil and blood lipids by comparison of the fatty acid composition of polar glycerolipids obtained by selective methanolysis without isolation from total lipids with that of polar lipids obtained by conventional methods.…”

Section: Discussionmentioning

confidence: 99%

Preparation of Fatty Acid Methyl Esters by Selective Methanolysis of Polar Glycerolipids

Ichihara

Yamaguchi

Araya

et al. 2010

Lipids

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KOH in aqueous methanol catalyzes selective methanolysis of polar glycerolipids with O-ester-linked acyl residues, while triacylglycerols and sterol esters are inert in the solution. Based on these findings, a convenient and reliable method was developed for the preparation of fatty acid methyl esters (FAMEs) from polar glycerolipids in lipid mixtures without prior isolation. Methanolysis of polar glycerolipids was completed within 2.5 min by vortexing or 20 min by shaking with 0.7 M KOH/70% (v/v) methanol in the presence of hexane at 30 degrees C. The yields of FAMEs obtained by the present method were greater than 95%. The method was applied successfully to gas chromatographic analysis of the fatty acid compositions of polar glycerolipids in seed oil and blood. No obvious differences were found between the fatty acid compositions determined by the present method and those determined by conventional methods, including lipid extraction with chloroform/methanol followed by isolation of polar lipids by chromatography. The fatty acid composition of polar glycerolipids, including phospholipids, can be determined readily in many crude samples.

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Section: Discussionmentioning

confidence: 99%

Preparation of Fatty Acid Methyl Esters by Selective Methanolysis of Polar Glycerolipids

Ichihara

Yamaguchi

Araya

et al. 2010

Lipids

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“…Dried cellular lipid extracts or individual lipid fractions were subjected to alkaline transesterification with methanol modified after (65). For this, dried lipids were resuspended in methanol/toluene (1:1, v/v), mixed with an equal volume of methanolic KOH and incubated for 1 h at 50 °C.…”

Section: Methodsmentioning

confidence: 99%

Steryl ester formation and accumulation in steroid-degrading bacteria

Holert

Brown

Hashimi

et al. 2019

Preprint

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16Steryl esters (SEs) are important storage compounds in many eukaryotes and are often 17 prominent components of intracellular lipid droplets. Here we demonstrate that selected 18Actino-and Proteobacteria growing on sterols are also able to synthesize SEs and to 19 sequester them in cytoplasmic lipid droplets. We found cholesteryl ester (CE) formation in 20 members of the actinobacterial genera Rhodococcus, Mycobacterium, and Amycolatopsis as 21 well as several members of the proteobacterial Cellvibrionales order. CEs maximally 22 environmental stress conditions to increase their survival in environments with fluctuating 48 nutrient conditions (1). A much smaller subset of bacteria are oleaginous and accumulate 49 neutral lipids, namely triacylglycerols (TAGs) and wax esters (WEs), as a prominent class of 50 storage compounds. Although TAG and WE synthesis is rarely described in other bacteria, 51 their accumulation is widespread within the actinobacterial genera Streptomyces, Nocardia, 52

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“…CE is more hydrophobic and less soluble in methanol than TAG, and methanolysis of CE does not proceed under the conditions for TAG [25,26]. Various solvents have been used to increase the solubility of CE in methanol to enhance its reactivity.…”

Section: Methanolysis Of Lipid Classes Separated By Silica Gel Spe Comentioning

confidence: 99%

“…Various solvents have been used to increase the solubility of CE in methanol to enhance its reactivity. For example, good yields of FAME are obtained in 1 h at 37°C or at room temperature by reaction in diethyl ether/methyl acetate/methanol containing 1 M CH 3 ONa (50:1:1, v/v/v) or in methyl propionate/ methanol containing 0.84 M NaOH (2:3, v/v), respectively [25,26]. A mixture of CE eluate (3 ml of 1% methyl acetate in hexane), 1.5 ml of acetone, and 0.6 ml of 2 M CH 3 ONa/methanol formed a single homogenous phase of the solvents, and methanolysis was completed in 30 min at room temperature (Fig.…”

Section: Methanolysis Of Lipid Classes Separated By Silica Gel Spe Comentioning

confidence: 99%

Improved Methods for the Fatty Acid Analysis of Blood Lipid Classes

Ichihara

Yoneda

Takahashi

et al. 2011

Lipids

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Two improved methods have been developed for preparation of fatty acid methyl esters (FAME) from major O-ester lipid classes in blood, i.e., cholesterol ester, triacylglycerol, and glycerophospholipids. The methods involve simple operations, and use neither harmful solvents such as chloroform or benzene nor highly reactive volatile reagents such as acetyl chloride. The FAME synthesis reaction proceeds under mild temperature conditions. The methods include (1) extraction of lipids from 0.2 ml of blood with 0.2 ml of tert-butyl methyl ether and 0.1 ml of methanol, (2) separation of the total lipids into lipid classes using a solid-phase extraction column or thin-layer chromatography, and (3) methanolysis of each lipid class at room temperature or at 45 °C. In all the operations, solvent concentration is performed only once prior to gas-liquid chromatography (GC). No noticeable differences in composition determined by GC have been found between FAME prepared by the present methods and those prepared by a conventional method involving lipid extraction with chloroform/methanol. The mild reaction and simplified procedures of the present methods enabled safe and reproducible analysis of the fatty acid compositions of the major ester-lipid classes in blood.

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Preparation of FAME from sterol esters

Cited by 8 publications

References 5 publications

Preparation of Fatty Acid Methyl Esters by Selective Methanolysis of Polar Glycerolipids

Preparation of Fatty Acid Methyl Esters by Selective Methanolysis of Polar Glycerolipids

Steryl ester formation and accumulation in steroid-degrading bacteria

Improved Methods for the Fatty Acid Analysis of Blood Lipid Classes

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