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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“To assess the lipid content, 300 mg of liver and 700 mg of white muscle were pulverized with liquid nitrogen in a mortar, and then extracted with a 2:1 chloroform:methanol solution following the method of Bligh and Dyer [23].”
Method
Assessment of cooking methods and freezing on the nutritional value and health risks of heavy metals in four fish species consumed in Douala, Cameroon
Christophe, M. K. J., Marlène, Y., François, N. J. V., et al. (2024) · Heliyon · doi:10.1016/j.heliyon.2024.e28316 · Cited by 11
“Total lipid was determined by method described by Bligh and Dyer [27] .”
Method
Enhanced viability and membrane lipid stability of cholesterol-lowering probiotics through spray drying encapsulation with whey protein isolate-coconut oil complex
Ngamekaue, N., Dumrongchai, T., Rodklongtan, A., & Chitprasert, P. (2024) · Drying Technology · doi:10.1080/07373937.2024.2334854 · Cited by 11
“Fatty acids were extracted by a modified Bligh and Dyer [17] method.”
Method
Potential CO2 biofixation by microalgae strains for industrial application
Gharanjik, M. A., Najafpour, G. D., Jahanshahi, M., & Mohammadi, M. (2024) · International Journal of Environmental Science and Technology · doi:10.1007/s13762-024-05562-3 · Cited by 10
Carcass and meat characteristics of Nellore young bulls fed diet using cottonseed cake as a replacer of the forage fiber source
Arcanjo, A. H. M., Ítavo, L. C. V., Ítavo, C. C. B. F., Gomes, M. D. N. B., Nazário, C. E. D., Gurgel, A. L. C., Dias‐Silva, T. P., Santana, J. C. S., da Silva, M. G. P., Coelho, F. D. A., et al. (2024) · Scientific Reports · doi:10.1038/s41598-024-58738-9 · Cited by 14
Transcriptomic and lipidomic analysis of the differential pathway contribution to the incorporation of erucic acid to triacylglycerol during Pennycress seed maturation
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Microelements, Fatty Acid Profile, and Selected Biomarkers in Grass Carp (Ctenopharyngodon idella) Muscle Tissue: Seasonal Variations and Health Risk Assessment
Kováčik, A., Helczman, M., Árvay, J., Tomka, M., Šnirc, M., Jančo, I., Fik, M., Štefunková, N., Dupák, R., Harangozo, Ľ., et al. (2024) · Biological Trace Element Research · doi:10.1007/s12011-024-04190-9 · Cited by 12
“Lipids from dried dorsal muscle were extracted by modified methods, according to Bligh and Dyer [45] and Folch et al. [46].”
Effects of local farming practices on soil organic carbon content, enzymatic activities, and microbial community structure in semi-arid soils of Morocco
Jindo, K., Aroussi, O. E., de Vente, J., et al. (2024) · Frontiers in Soil Science · doi:10.3389/fsoil.2024.1369971 · Cited by 11
Activation of alveolar epithelial ER stress by β-coronavirus infection disrupts surfactant homeostasis in mice: implications for COVID-19 respiratory failure
Murthy, A., Rodríguez, L. R., Dimopoulos, T. T., Bui, S., Iyer, S., Chavez, K., Tomer, Y., Abraham, V., Cooper, C., Renner, D. M., et al. (2024) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00324.2023 · Cited by 11
Tailor-made packaging strategies of fresh pork belly with different fat content: Enhancing shelf life while minimizing environmental impact
Marcos, B., Quinteiro, P. S. G. N., Albano‐Gaglio, M., Muñoz, I., Claret, A., Guerrero, L. A. V., Lloret, E., Tejeda, J. F., & Font‐i‐Furnols, M. (2024) · Meat Science · doi:10.1016/j.meatsci.2024.109627 · Cited by 11
Effect of iron oxide nanoparticles on mixotrophic cultivation of Chlorella spp. for biofuel production
Rana, M. S., Ariyadasa, T. U., & Prajapati, S. K. (2024) · Bioresource Technology · doi:10.1016/j.biortech.2024.131241 · Cited by 11
Functional overlap of inborn errors of immunity and metabolism genes defines T cell metabolic vulnerabilities
Patterson, A. R., Needle, G. A., Sugiura, A., Jennings, E. Q., Chi, C., Steiner, K. K., Fisher, E. L., Robertson, G. L., Bodnya, C., Markle, J., et al. (2024) · Science Immunology · doi:10.1126/sciimmunol.adh0368 · Cited by 12
“Metabolites were extracted using Bligh-Dyer liquid-liquid extraction (1:1:0.9 chloroform:methanol:water, v/v) as reported previously at a concentration of 4 × 10 6 cells/ml ( 74 , 75 ).”
MethodInfluential
Mitigating gadolinium toxicity in guar (Cyamopsis tetragonoloba L.) through the symbiotic associations with arbuscular mycorrhizal fungi: physiological and biochemical insights
AbdElgawad, H., Crecchio, C., Nhs, M., Abdel-Maksoud, M. A., Malik, A. R. A., Sheteiwy, M. S. A., Hamoud, Y. A., Sulieman, S., Shaghaleh, H., Alyafei, M. A. M., et al. (2024) · BMC Plant Biology · doi:10.1186/s12870-024-05552-0 · Cited by 11
Loss of adipose ATF3 promotes adipose tissue lipolysis and the development of MASH
Hu, S., Bawa, F. N. C., Zhu, Y., Pan, X., Wang, H., Gopoju, R., Xu, Y., & Zhang, Y. (2024) · Communications Biology · doi:10.1038/s42003-024-06915-x · Cited by 12
“Analysis of lipids, hydroxyproline, ROS, adiponectin, leptin, insulin, AST, and ALT Liver tissue (100mg) was homogenized in methanol, and lipids were extracted in chloroform/methanol (2:1v/v) as described 47 .”
Effects of salinity and temperature on growth performance, biochemical composition, and biosilification process of Cyclotella cryptica
Zhao, Y., Sun, Y., Zhu, Z., Li, Y., Zhang, L., Li, J., Agathos, S. N., Zhou, C., & Han, J. (2024) · Algal Research · doi:10.1016/j.algal.2024.103751 · Cited by 10
Exploring the response of yellow lupine (Lupinus luteus L.) root to drought mediated by pathways related to phytohormones, lipid, and redox homeostasis
Burchardt, S., Czernicka, M., Kućko, A., et al. (2024) · BMC Plant Biology · doi:10.1186/s12870-024-05748-4 · Cited by 11
“Total lipids were extracted according to a modified method described by Bligh and Dyer [23].”
Method
Effect of ultrasound pre-treatment prior to enzymatic hydrolysis on physicochemical parameters of fish protein hydrolysates (FPH) extracted from side streams of Atlantic mackerel (Scomber scombrus)
Cropotova, J., Kvangarsnes, K., Stangeland, J., & Rustad, T. (2024) · Frontiers in Sustainable Food Systems · doi:10.3389/fsufs.2024.1500359 · Cited by 13
Effect of High-Pressure Processing on Color, Texture and Volatile Profile During Sardine Refrigeration
Ismaiel, L., Nartea, A., Fanesi, B., et al. (2025) · Foods · doi:10.3390/foods14020329 · Cited by 11
Optimization of co-valorisation techniques for dairy and paper pulp wastewater in the cultivation of Chlorococcum sp. with a focus on mixture design, microwave-assisted pretreatment, and bioethanol production
Ngerem, E. C., Sanusi, I. A., Kana, G. E. B., & Olaniran, A. O. (2025) · Heliyon · doi:10.1016/j.heliyon.2025.e42531 · Cited by 13
“Lipid content was determined using the Bligh and Dyer [31] solvent extraction method.”
Hydrolyzed collagen, KCl, and arginine: A successful strategy to reduce fat and sodium while maintaining the physicochemical, sensory, and shelf life quality of mortadella
Correa, L. P., Pinton, M. B., dos Santos, B. A., Morgano, M. A., Vargas‐Ramella, M., Cichoski, A. J., Wagner, R. C., & Campagnol, P. C. B. (2025) · Meat Science · doi:10.1016/j.meatsci.2025.109775 · Cited by 11
Cinnabarinic acid protects against metabolic dysfunction-associated steatohepatitis by activating aryl hydrocarbon receptor-dependent AMPK signaling
Patil, N. Y., Rus, I., Ampadu, F. A., Abushukair, H. M., Bonvicino, S. M., Brush, R. S., Eaton, E., Agbaga, M., Oh, T. G., Friedman, J. E., et al. (2025) · American Journal of Physiology-Gastrointestinal and Liver Physiology · doi:10.1152/ajpgi.00337.2024 · Cited by 11
Imaging and spatially resolved mass spectrometry applications in nephrology
Gorman, B. L., Shafer, C. C., Ragi, N., Sharma, K., Neumann, E. K., & Anderton, C. (2025) · Nature Reviews Nephrology · doi:10.1038/s41581-025-00946-1 · Cited by 19
“Although no standardized methods of extracting analytes exist, many lipidomics studies have adapted the Bligh and Dyer method for lipid extraction 70 .”
Application of plant-derived extracellular vesicles as novel carriers in drug delivery systems: a review
Yang, J., Ai, X., Zhang, C., Guo, T., & Feng, N. (2025) · Expert Opinion on Drug Delivery · doi:10.1080/17425247.2025.2487589 · Cited by 15
“Methods by Bligh and Dyer (Figure 6(b)) were generally used to extract total lipids from P-EVs [104].”
Dietary targeting of TRPM8 rewires macrophage immunometabolism reducing colitis severity
Cicia, D., Biscu, F., Iannotti, F. A., Miraglia, M., Ferrante, C., Iaccarino, N., Miguel, S. C.-D., Chiavaroli, A., Moriello, A. S., Cicco, P. D., et al. (2025) · Cell Death and Disease · doi:10.1038/s41419-025-07553-9 · Cited by 12
“The chromatographic gradient was run at a ﬂ ow rate of 0.150 ml/min as follows: 0 – 20 min: linear gradient from 80% to 20% B; 20 – 20.5 min: linear gradient from 20% to 80% B; 20.5 – 28 min: hold at 80% B.”
Influential
Cell disruption and extraction for microalgae biorefinery: Focus on deep eutectic solvents moving towards aqueous biphasic system
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Genotype, production system and sex effects on fatty acid composition of meat from goat kids
Özcan, M., Demirel, G., Yakan, A., et al. (2014) · Animal Science Journal · doi:10.1111/asj.12273 · Cited by 11
Improvement of lipid stability of refrigerated rainbow trout (Oncorhynchus mykiss) fillets by pre-storage α-tocopherol acetate dipping treatment.
Ehsani, A., & Jasour, M. S. (2012) · PubMed · Cited by 11
Artemisia capillaris and Paecilomyces japonica Stimulate Lipid Metabolism and Reduce Hepatotoxicity Induced by Carbon Tetrachloride in Rats
Jin, Y.-X., 유윤숙, 한은경, et al. (2008) · Journal of the Korean Society of Food Science and Nutrition · doi:10.3746/jkfn.2008.37.5.548 · Cited by 11
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Bae, J.-H., Yoon, S.-H., & Lim, S. (2010) · Korean Journal of Fisheries and Aquatic Sciences · doi:10.5657/kfas.2010.43.1.006 · Cited by 11
Comparison of docosahexaenoic acid uptake in murine cardiomyocyte culture and tissue: Significance to physiologically relevant studies
Lamazière, A., Richard, D., Bausero, P., et al. (2014) · Prostaglandins Leukotrienes and Essential Fatty Acids · doi:10.1016/j.plefa.2014.11.004 · Cited by 11
Characterization of the growth, biochemical composition, and nutrient utilization of the cyanobacterium Geitlerinema lemmermannii
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Utilization of Soluble Starch by Oleaginous Red Yeast Rhodotorula glutinis
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Kodan, A., Shibata, H., Matsumoto, T., et al. (2009) · Protein Expression and Purification · doi:10.1016/j.pep.2009.02.010 · Cited by 11
Pretreatment for Simultaneous Production of Total Lipids and Fermentable Sugars from Marine Alga, Chlorella sp.
Lee, C.-G., Kang, D.-H., Lee, D.-B., & Lee, H.-Y. (2013) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-013-0295-y · Cited by 11
Quality Changes of Marinated Tench (Tinca tinca) during Refrigerated Storage
Özoğul, Y., Küley, E., & Özoğul, F. (2009) · Food Science and Technology International · doi:10.1177/1082013209350541 · Cited by 11
The Composition of fatty acids and aldehydes of the marine bryozoans Berenicea meandrina and Dendrobeania flustroides (Bryozoa: Gymnolaemata)
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Influence of Ice Storage on Raw Materials for the Production of High Quality Dried Fish Products
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Respiratory Deficiency Mediates the Regulation of CHO1-encoded Phosphatidylserine Synthase by mRNA Stability in Saccharomyces cerevisiae
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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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