Cited by 54,751Lipid metabolism and biosynthesisEdible Oils Quality and AnalysisFood Nutrient Composition and Product Development

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A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATION

Bligh, E. G., Dyer, W. J.·1959·Canadian Journal of Biochemistry and Physiology

DOI 10.1139/o59-099

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Abstract

Lipid decomposition studies in frozen fish have led to the development of a simple and rapid method for the extraction and purification of lipids from biological materials. The entire procedure can be carried out in approximately 10 minutes; it is efficient, reproducible, and free from deleterious manipulations. The wet tissue is homogenized with a mixture of chloroform and methanol in such proportions that a miscible system is formed with the water in the tissue. Dilution with chloroform and water separates the homogenate into two layers, the chloroform layer containing all the lipids and the methanolic layer containing all the non-lipids. A purified lipid extract is obtained merely by isolating the chloroform layer. The method has been applied to fish muscle and may easily be adapted to use with other tissues.

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A SIMPLE METHOD FOR THE ISOLATION AND PURIFICATION OF TOTAL LIPIDES FROM ANIMAL TISSUES
Folch, J., Lees, M., & Stanley, G. H. S. (1957) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)64849-5 · Cited by 65,189
PREPARATION OF LIPIDE EXTRACTS FROM BRAIN TISSUE
Folch, J., Ascoli, I., Lees, M. B., Meath, J. A., & Lebaron, F. N. (1951) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)55987-1 · Cited by 1,341
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Dyer, W. J., & Morton, M. L. (1956) · Journal of the Fisheries Research Board of Canada · doi:10.1139/f56-007 · Cited by 69
Self-citation
Experimental Determination of Binodal Curves, Plait Points, and Tie Lines, in 50 Systems, each consisting of Water and Two Organic Liquids
Bonner, W. D. (1910) · The Journal of Physical Chemistry · doi:10.1021/j150116a004 · Cited by 46
Acetone–Water Mixtures for the Extraction and Rapid Estimation of Fats of Biological Materials, Particularly Fish Products
Dambergs, N. (1956) · Journal of the Fisheries Research Board of Canada · doi:10.1139/f56-045 · Cited by 18

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BANCROFT W., 1895, Phys. Iiev., 3, 1

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Altın, N., & Uyar, B. (2025) · International Journal of Green Energy · doi:10.1080/15435075.2025.2459134 · Cited by 1
“Bligh and Dyer method was applied to determine the total lipid content and to extract lipids from microalgae (Bligh and Dyer 1959).”
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Pitt, A., Lienbacher, S., Schmidt, J., et al. (2025) · International Microbiology · doi:10.1007/s10123-025-00642-x · Cited by 1
“The polar lipids were extracted and analyzed by thin-layer chromatography according to the methods of Tindall (Tindall 1990a, 1990b), which was based on the description by Bligh and Dyer (Bligh and Dyer 1959).”
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Minimal changes in microbial abundances and diversity over 7 years of emplacement for modules of compacted bentonite exposed to natural groundwater
Sidhu, H. S., Engel, K., Ford, S. E., Keech, P. G., Behazin, M., Binns, W. J., Srikanthan, N., Simpson, M. J., Neufeld, J. D., & Slater, G. F. (2025) · Applied and Environmental Microbiology · doi:10.1128/aem.01950-24 · Cited by 1
“Briefly, samples (outer layer 1.6 to 6.9 g; inner layer 22.2–95.3 g) were crushed and lipids were extracted using a modified Bligh and Dyer protocol (50, 51) where an excess of dichloromethane-metha nol-phosphate buffer (1:2:0.8 vol/vol) was used as extraction solvent.”
Influential
Campylobacter jejuni resistance to human milk involves the acyl carrier protein AcpP
Zhou, B., Garber, J. M., Butcher, J., Muszyński, A., Casey, R. L., Huynh, S., Archer‐Hartmann, S. A., Porfírio, S., Rogers, A. M., Azadi, P., et al. (2025) · mBio · doi:10.1128/mbio.03997-24 · Cited by 1
“Bacteria pellets were extracted using the Bligh and Dyer method, as described previously (61, 62).”
Ecological physiology of recovery from experimental phenology of algal food deprivation in larvae of a sea urchin (Lytechinus pictus)
Griffith, A. W., Li, N., & Manahan, D. T. (2025) · Marine Biology · doi:10.1007/s00227-025-04597-5 · Cited by 1
“Lipids from tissue homogenates were first extracted in chloroform-methanol following standard methods described by Bligh and Dyer (1959 ), as modified by Holland and Gabbott ( 1971), and Moran and Manahan (2004) with appropriate use of internal standards.”
Production and characterization of a functional pasta for individuals with gluten-related disorders
Yerlikaya, P., Yatmaz, H. A., Erem, F., et al. (2025) · British Food Journal · doi:10.1108/bfj-11-2024-1148 · Cited by 1
Differentiated fatty acid allocation of Daphnia magna helped to maintain their population under food quality deterioration
Wang, S., Liu, Z., Su, X., et al. (2025) · Frontiers in Microbiology · doi:10.3389/fmicb.2025.1544005 · Cited by 1
“…Scenedesmus , Microcystis (3–5 mg), and D. magna (2–3 mg) samples were extracted according to the method described in a previous study by Bligh and Dyer (1959) and measured using gas chromatography–mass spectrometry (TRACE GC–MS, Thermo Finnigan, USA) at Analytical and Testing Center at…”
Technology for obtaining linseed protein concentrate minimized in antinutritional factors
Cargnin, G., Adorian, T. J., Falcone, D. B., et al. (2025) · Tropical Animal Health and Production · doi:10.1007/s11250-025-04344-4 · Cited by 1
“923.03), crude protein (method nº 960.52), total, soluble and insoluble dietary fiber (method nº 991.43) as described by AOAC (1995), lipids according to Bligh and Dyer (1959).”
Influential
BIOETHANOL PRODUCTION FROM HYDROLYSATE DERIVED BY ULTRASONIC PRETREATED DEFATTED BIOMASS OF MUNICIPAL WASTEWATER GROWN MUTANT TETRADESMUS DIMORPHUS EMS2
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“Bligh and Dyer’s [23] method was adapted for lipid extraction from dried biomass.”
Subcellular Fractionation Enables Assessment of Nucleotide Sugar Donors Inside the Golgi Apparatus as a Prerequisite for Unraveling Culture Impacts on Glycoforms of Antibodies
Regett, N., Dieterle, M., Peters, F., Deuring, M., Stegmaier, K., Teleki, A., & Takors, R. (2025) · Biotechnology Journal · doi:10.1002/biot.202400678
“Metabolic extraction was performed using a modified version of the procedure developed by Bligh and Dyer [36].”
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“Lipid was extracted by homogenising the sample with a mixture of chloroform and methanol, as described by [35], and the fatty acid composition was analysed using the improved method, where fatty acids were esterified with methanol in sodium hydroxide and catalysed by borontrifluride.”
Comparative analysis of different catalysts in conventional and In-Situ transesterification for enhanced biodiesel yield and quality from Scenedesmus obliquus
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“One of the most widely used lipid extraction techniques is the Bligh-Dyer method [23], it includes a combination of polar and non-polar solvents to maximize lipid recovery [24].”
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“Lipid content was analyzed using the Bligh and Dyer method ( 18 ).”
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“As the widely used lipid extraction methods (Folch et al. 1957, Bligh and Dyer 1959) are time-consuming, various simplified extraction methods have been developed (Abdulkadir and Tsuchiya 2008).”
Energy allocation strategy during the reproductive cycle of Patagonian grouper (Acanthistius patachonicus) from the southwestern Atlantic Ocean
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“Lipids were extracted using the method of Bligh and Dyer (1959) modified by Undeland et al. (1998), and quantified gravimetrically following the procedure outlined by Herbes and Allen (1983).”
Description of Labrys sedimenti sp. nov., isolated from a diclofenac-degrading enrichment culture, and genome-based reclassification of Labrys portucalensis as a later heterotypic synonym of Labrys neptuniae
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“The polar lipids and respiratory quinones were extracted based on the protocol modified after Bligh and Dyer [36].”
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Arslan, Ş., Abdelsalam, A. H., Kurt‐Kızıldoğan, A., Otur, Ç., Konuş, M., Çetin, D., Babat, C. F., Yılmaz, C., Oğuztüzün, S., Sayın, S., et al. (2025) · Biology Bulletin · doi:10.1134/s1062359024608814 · Cited by 1
“In addition, lipid content and fatty acid analysis of extract were made according to the method described by Bligh and Dyer (1959) and Garches and Mancha (1993), respectively.”
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Polar microalgae-based nanoparticles: a next-generation strategy for cancer treatment
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“(3) (Bligh and Dyer 1959).”

Authors: Bligh, E. G., & Dyer, W. J.
Venue: Canadian Journal of Biochemistry and Physiology
Publisher: Canadian Science Publishing
Published: 1959-08-01
Volume: 37
Issue: 8
Pages: 911-917
ISSN: 0576-5544 (ISSN-L)
DOI: 10.1139/o59-099
OpenAlex: W2152311269
Cited by: 54,751
References: 5
Citations by year: 2026: 1149 · 2025: 1534 · 2024: 1685 · 2023: 1749 · 2022: 1802 · 2021: 2073 · 2020: 2152 · 2019: 1870 · 2018: 1737 · 2017: 1784 · 2016: 1780 · 2015: 1800 · 2014: 1786 · 2013: 1758 · 2012: 1551
FWCI: 8.73
Type: journal article
Language: en
Open access: closed
Licence: www.nrcresearchpress.com/page/about/CorporateTextAndDataMining (tdm)
Locations: Canadian Journal of Biochemistry and Physiology (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
Physical Sciences › Chemistry › Organic Chemistry › Edible Oils Quality and Analysis
Life Sciences › Agricultural and Biological Sciences › Food Science › Food Nutrient Composition and Product Development
Keywords: lipid extraction, lipid purification, chloroform-methanol extraction, frozen fish, lipid degradation, fish muscle, tissue homogenization, rapid extraction method
MeSH terms: Fishes, Lipids

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A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATION

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