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
Storage of Frozen Plaice Fillets
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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Thangaraj, S. V., Ghnenis, A. B., Pallas, B. D., et al. (2024) · Scientific Reports · doi:10.1038/s41598-024-58116-5 · Cited by 4
“Lipids were extracted using a modified Bligh-Dyer method 55 .”
Method
Waterborne exposure to the antineoplastic 5-fluorouracil alters lipid composition in larval zebrafish (Danio rerio)
Ivantsova, E., Henry, E., Konig, I. F. M., et al. (2024) · The Science of The Total Environment · doi:10.1016/j.scitotenv.2024.172377 · Cited by 4
Photosynthetic carbon allocation in native and invasive salt marshes undergoing hydrological change
Li, Y., Xie, L., Li, S., et al. (2024) · The Science of The Total Environment · doi:10.1016/j.scitotenv.2024.173232 · Cited by 4
Fatty Acid Composition and Anticancer Activity of Neutral and Polar Lipids of Pacific Oyster (Crassostrea gigas) Cultured in Khanh Hoa Coast in Vietnam
Nguyen, M. V., Kakooza, D., Do, T. H. T., Tran, A. P. T., Nguyen, T. H., & Tran, N. Q. (2024) · Polish Journal of Food and Nutrition Sciences · doi:10.31883/pjfns/188139 · Cited by 4
“Human cell lines, including MDA-MB-231 (human breast cancer), HepG2 (liver cancer), and RD (muscle rhabdomyosarcoma-A), were procured from ATCC (Manassas, VA, USA). r Lipid extraction and fractionation Total lipids were extracted from oyster muscle following the method of Bligh & Dyer [1959].”
Method
In Vitro Multi-Bioactive Potential of Enzymatic Hydrolysis of a Non-Toxic Jatropha curcas Cake Protein Isolate
Olloqui, E. J., Alejandro, G.-R. M., Elizabeth, C.-L., et al. (2024) · Molecules · doi:10.3390/molecules29133088 · Cited by 4
“The resulting residues were ground, sieved, and later defatted using the Bligh–Dyer methodology [27] to obtain Jatropha curcas cake.”
Method
The brominated flame retardant hexabromocyclododecane causes systemic changes in polyunsaturated fatty acid incorporation in mouse lipids
Kramer, N. E., Siracusa, J. S., Xu, H., Barnett, L. M. A., Finnerty, M. C., Guo, T. L., Wagner, J. J., Leach, F. E., & Cummings, B. S. (2024) · Toxicological Sciences · doi:10.1093/toxsci/kfae094 · Cited by 5
Water restriction as a strategy for growing Talinum fruticosum (L.) Juss. (Talinaceae)
Santos, R. D. J., do Nascimento, M. N., Camilloto, G. P., et al. (2024) · Revista Caatinga · doi:10.1590/1983-21252024v3712183rc · Cited by 4
The impact of drying and extraction methods on total lipid, fatty acid profile, and cytotoxicity of Tenebrio molitor larvae
Abutaha, N., & Al-Mekhlafi, F. A. (2024) · Open Chemistry · doi:10.1515/chem-2024-0110 · Cited by 8
Assessing the efficiency of ethyl acetate for lipid extraction as an alternative to the Folch method in selected marine low‐trophic species
Wang, Y., Eilertsen, K., Elvevoll, E. O., & Walquist, M. J. (2025) · Journal of the American Oil Chemists Society · doi:10.1002/aocs.12935 · Cited by 5
The behavioral, physiological, and biochemical responses of Lumbriculus variegatus exposed to cannabidiol and its metabolites
Williams, B. S., Jomy, G., Flanagan, M., et al. (2025) · Environmental Toxicology and Chemistry · doi:10.1093/etojnl/vgaf048 · Cited by 4
“The Bligh and Dyer (1959) method was utilized for lipid extraction with extraction conducted by resuspension of samples in 300 µ l deionized water with samples then transferred to 2ml glass gas chromatography (GC) vials.”
Recovery of Lipid Biomarkers in Hot Spring Digitate Silica Sinter as Analogs for Potential Biosignatures on Mars: Results from Laboratory and Flight-Like Experiments
Millan, M., Campbell, K. A., Sriaporn, C., Handley, K. M., Teece, B. L., Mahaffy, P., & Johnson, S. S. (2025) · Astrobiology · doi:10.1089/ast.2024.0020 · Cited by 4
“…a range of methods exist for lipids extraction (Saini et al., 2021), the lipids in this study were extracted using a modi ﬁ ed Bligh & Dyer method (Bligh and Dyer, 1959), and an extraction blank was performed on combusted sand to monitor any potential contamination of the whole extraction and…”
Dentiradicibacter hellwigii gen. nov., sp. nov., isolated from a secondary infected root canal in the human oral cavity
Bartsch, S., Wittmer, A., Weber, A.-K., et al. (2025) · INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY · doi:10.1099/ijsem.0.006690 · Cited by 4
“Polar lipids were from wet biomass using a modified Bligh and Dyer extraction [42].”
Algal nanobionics and a nanobiohybrid system of biosynthesized ZnO-NPs to augment the cellular accumulation of biomolecules and its effect on microalgae
Parveen, A., Rawat, J., Bisht, B., et al. (2025) · New Journal of Chemistry · doi:10.1039/d4nj03976a · Cited by 4
Overexpression of Plastid Acetyl-CoA Carboxylase Confers Stress Tolerances with Increased Levels of Unsaturated Fatty Acids in the Marine Diatom Phaeodactylum tricornutum
Toyoshima, M., Okuda, H., Okada, Y., Yoneda, K., Shimakawa, G., & Matsuda, Y. (2025) · Marine Biotechnology · doi:10.1007/s10126-025-10446-9 · Cited by 5
“Total lipids were extracted by the method of Bligh and Dyer (1959).”
Liver gene expression and its rewiring in hepatic steatosis are controlled by PI3Kα-dependent hepatocyte signaling
Régnier, M., Polizzi, A., Fougeray, T., Fougerat, A., Perrier, P., Anderson, K. E., Lippi, Y., Smati, S., Lukowicz, C., Lasserre, F., et al. (2025) · PLoS Biology · doi:10.1371/journal.pbio.3003112 · Cited by 4
“…samples were homogenized in methanol/5 mM EGTA (2:1, v/v) and lipids (equivalent to 2 mg of tissue) extracted according to the Bligh–Dyer method [43] with chloroform/methanol/water (2.5:2.5:2 v/v/v) in the presence of the following internal standards: glyceryl trinonadecanoate, stigmasterol,…”
Influential
Bioactive Metabolites from Radioresistant Bacterium Kocuria sp. TMM 11 and Their Role in Prevention of Ultraviolet-Induced Photodamages
Alam, T., Din, S. U., Abdullah, M., Ali, M., Badshah, M., Farman, M., Khan, S., Hasan, F., & Shah, A. A. (2025) · Current Microbiology · doi:10.1007/s00284-025-04194-1 · Cited by 4
“Lipid was extracted using the standard protocol as described previously [26].”
Protein hydrolysates from Italian sea bream and sea bass aquaculture side streams – chemical, biological, and techno-functional characterization
Jenssen, M., Matić, J., Sone, I., Solberg, N. T., Wubshet, S. G., & Lian, K. (2025) · Frontiers in Marine Science · doi:10.3389/fmars.2025.1572015 · Cited by 5
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Dong, P., Zhang, Y., Liu, C., Yan, D., Xu, X. P., Li, A., & Jin, L.-M. (2025) · Journal of Nutrition · doi:10.1016/j.tjnut.2025.05.022 · Cited by 4
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Cesari, A. B., Paulucci, N. S., & Dardanelli, M. S. (2025) · Plants · doi:10.3390/plants14111601 · Cited by 4
“Total lipids from bacteria immobilized from T0 and T6 beads A, B, and C, were extracted according to the method of [35].”
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Tapia-García, E. Y., Chávez‐Ramírez, B., Larios‐Serrato, V., et al. (2025) · PLoS ONE · doi:10.1371/journal.pone.0324390 · Cited by 4
“The strains were grown in LB liquid media for 16 h at 29°C, and the polar lipids were extracted following the Bligh and Dyer technique [37].”
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Varadharasu, P., Dhandayuthapani, K., Annadurai, P., Ahuja, V., Chauhan, S., Tilak, K., Sundarrajan, B., Abdi, G., & Sharma, D. (2025) · Scientifica · doi:10.1155/sci5/8055358 · Cited by 5
“Total sugar and lipid content in extract were assessed by the DuBois method [25] and gravimetric method [26], respectively.”
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“The lipid fraction of native cells was isolated using a chloroform/isopropanol mixture, followed by chloroform alone, based on a modified Bligh and Dyer 69 method.”
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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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