Comparison of the bligh and dyer and folch methods for total lipid determination in a broad range of marine tissue

Abstract

For many studies, it is important to measure the total lipid content of biological samples accurately. The Bligh and Dyer method of extraction was developed as a rapid but effective method for determining total lipid content in fish muscle. However, it is also widely used in studies measuring total lipid content of whole fish and other tissues. Although some investigators may have used modified Bligh and Dyer procedures, rarely have modifications been specified nor has their effectiveness been quantitatively evaluated. Thus, we compared this method with that of the classic Folch extraction in determining total lipid content of fish samples ranging from 0.5 to 26.6% lipid. We performed both methods as originally specified, i.e., using the chloroform/methanol/water ratios of 1:2:0.8 and 2:2:1.8 (before and after dilution, respectively) for Bligh and Dyer and of 8:4:3 for Folch, and with the initial solvent/sample ratios of (3+1):1 (Bligh and Dyer) and 20:1 (Folch). We also compared these with several other solvent/sample ratios. In samples containing <2% lipid, the results of the two methods did not differ. However, for samples containing >2% lipid, the Bligh and Dyer method produced significantly lower estimates of lipid content, and this underestimation increased significantly with increasing lipid content of the sample. In the highest lipid samples, lipid content was underestimated by up to 50% using the Bligh and Dyer method. However, we found a highly significant linear relationship between the two methods, which will permit the correction of reported lipid levels in samples previously analyzed using an unmodified Bligh and Dyer extraction. In the future, modifications to procedures and solvent/sample ratios should be described.

Sterol and genomic analyses validate the sponge biomarker hypothesis

“…Briefly, lipid extractions were performed using a modified Bligh-Dyer method (44). The total lipid extracts (TLEs) were saponified by mild alkaline methanolysis, generating fatty-acid methyl-esters (45).…”

Recent developments in microbial oils production: a possible alternative to vegetable oils for biodiesel without competition with human food?

“…The aqueous superior phase contains only traces of lipids while the lower phase contains the extracted lipids. The upper phase is rinsed with the mixture of chloroform, methanol and water (organic solvent) in a ratio of 86:14:1 to remove the traces of lipids while the lower phase is rinsed with the mixture of chloroform, methanol and water (aqueous solvent) in a ratio of 3:48:47 to remove the non-lipid contaminants (Folch et al 1957;Iverson et al 2001). The organic phase used for the rinsing of upper phase may be added with salts which help to eliminate lipids from the upper phase.…”

“…After rinsing the organic phases are pooled and evaporated to dryness and the residue is weighed. This method has been adapted or modified many times (Iverson et al 2001). The well known modification is that of Bligh and Dyer (1959) where the sample to solvent ratio has been lowered to 3, allowing a reduction of solvent utilisation and warranting more than 95% of lipid recovery.…”

The antioxidative effect of lipophilized rutin and dihydrocaffeic acid in fish oil enriched milk

“…Lipids were extracted from fish oil enriched milk (15 g) according to the method described by Bligh and Dyer [21] with reduced amount of solvent applied [22]. The analysis was done in duplicate and further used for determination of peroxide value, fatty acid composition and tocopherol concentration.…”