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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“Then, the lipid was extracted from dried algae biomass by following previously reported method [24].”
Comparative efficacy of different tenderizing agents and time on Physicochemical textural and organoleptic properties of squid (Todarodes pacificus) muscle
Baykal, M., Çelik, M., Çelik, L., et al. (2023) · Food Science & Nutrition · doi:10.1002/fsn3.3751 · Cited by 4
“Lipid content was assayed as per Bligh and Dyer (1959); and pH analyses were done according to the method of Lima Dos Santos et al. (1981).”
Method
Microalgae Biomass and Bioactive Compounds Change According to the Medium's N and pH
da Silva, H. R., Prete, C. E. C., Bertoldo, L. A., Basso, E. A., de Melo, U. Z., Nassar, J. M., & Andrade, D. S. (2023) · Brazilian Archives of Biology and Technology · doi:10.1590/1678-4324-2023210415 · Cited by 4
“Lipid determination was based on the gravimetry method [33], as described by Ryckebosch et al. [34], in which total lipids were extracted from 50 mg of lyophilized biomass with chloroform:methanol:water at a ratio of 1:1:0.8.”
MethodInfluential
Significant influence of four highly conserved amino-acids in lipochaperon-active sHsps on the structure and functions of the Lo18 protein
Bellanger, T., Barreira, D. D. S., Wien, F., et al. (2023) · Scientific Reports · doi:10.1038/s41598-023-46306-6 · Cited by 4
Spatiotemporal variation of microbial communities in surficial sediments of Cochin estuary, southwest coast of India
Joseph, N., Jagadeesan, L., Jangid, K., et al. (2023) · Environmental Monitoring and Assessment · doi:10.1007/s10661-023-12023-w · Cited by 4
“Total LPD was extracted using chloroform and methanol for elution and estimated according to Bligh and Dyer (1959) and Parsons et al. (1984) using stearic acid as the standard.”
Method
Prebiotic Activity of Pequi ( Caryocar brasiliense Camb.) Shell on Lactobacillus and Bifidobacterium Strains: A Medicinal Food Ingredient
Alves, A. M., Silva, M. M. D. A., Rodrigues, C. A. P., de Albuquerque, T. M. R., de Souza, E. L., & Naves, M. M. V. (2023) · Journal of Medicinal Food · doi:10.1089/jmf.2022.0117 · Cited by 4
Energy transfers in a predator–prey context involving D. salina (microalga), F. salina (ciliate) and A. salina (crustacean), living in salterns of Sfax (Tunisia)
Guermazi, W., Athmouni, K., Annabi-Trabelsi, N., Elloumi, J., Ayadi, H., & Leignel, V. (2024) · Aquatic Ecology · doi:10.1007/s10452-023-10082-2 · Cited by 6
“Total lipids of all studied species were extracted with a mixture of chloroform–methanol (2:1, v/v) according to the method of (Bligh and Dyer 1959).”
Preferential phosphatidylglycerol synthesis via phosphorus supply through rRNA degradation in the cyanobacterium, Synechocystis sp. PCC 6803, under phosphate-starved conditions
Hiyoshi, T., Haga, M., & Sato, N. (2024) · Frontiers in Plant Science · doi:10.3389/fpls.2024.1335085 · Cited by 4
Lipidomics Analysis Deepen Understanding the Molecular Mechanisms in a Gouty Model Induced by Combination of MSU Crystals Injection and High-Fat Diet Feeding and the Intervention Mechanisms of Allopurinol
Li, D., Shi, W., Hu, C., et al. (2024) · Journal of Inflammation Research · doi:10.2147/jir.s443358 · Cited by 4
“18,19 Each lipid extract was resuspended with 200 μL chloroform/methanol (1:1, v/v)/100 uL serum, and stored at −20 ◦C for lipid analysis.”
Method
A comparative analysis of cutin monomers from cereal brans and fruit peels: Isolation and characterization
Li, Y., Yadav, M. P., Moreau, R. A., et al. (2024) · Journal of Cereal Science · doi:10.1016/j.jcs.2024.103873 · Cited by 4
A progesterone derivative linked to a stable phospholipid activates breast cancer cell response without leaving the cell membrane
Font-Mateu, J., Sanllehí, P., Sot, J., Abad‐García, B., Mateos, N., Torreno-Pina, J. A., Ferrari, R., Wright, R. H. G., Parajo, M. F. G., Joglar, J., et al. (2024) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-024-05116-3 · Cited by 4
“The Bligh and Dyer protocol was applied to extract the lipidic fraction [40].”
Method
Proximate and fatty acids composition in the muscle of wild and farmed sardine (Sardinella brasiliensis)
Scheuer, F., Sterzelecki, F. C., Wagner, R. C., et al. (2024) · Food Chemistry Advances · doi:10.1016/j.focha.2024.100637 · Cited by 4
Nutraceutical potential of microalgae: a case study from a tropical estuary in Southern India
Sandeep, K. P., Sivaramakrishnan, T., Raymond, J. A. J., Sudheer, N. S., Thomas, D., Dayal, J. S., Balasubramanian, C. P., Kailasam, M., & Ambasankar, K. (2024) · Journal of Coastal Conservation · doi:10.1007/s11852-024-01046-1 · Cited by 4
“The lipid content was estimated by the Bligh and Dyer method (Bligh and Dyer 1959), and the respective fatty acid methyl esters (FAMEs) were prepared and extracted into petroleum ether.”
Idiosyncratic Fermentation Behaviour of Rhodosporidium toruloides NCIM 3547 in Hemicellulose Hydrolysates Derived from Neem Oilseed Cake for Lipid and β-carotene Synthesis
Bharathi, S. D., & Jacob, S. (2024) · Waste and Biomass Valorization · doi:10.1007/s12649-024-02441-3 · Cited by 4
“Bligh and Dyer’s method was followed with slight modifications for extracting and estimating lipids from the biomass produced using five different media [39].”
A rare urinary tract infection of multidrug-resistant Chryseobacterium urinae sp. nov. isolated from a diabetic, non-catheterized patient
Bhatt, A., Pujari, S., Mantri, S., et al. (2024) · Archives of Microbiology · doi:10.1007/s00203-024-03881-0 · Cited by 4
“A mixture of Methanol/Chlo-roform/0.3% Sodium Chloride (2:1:0.8, by vol.) facilitated polar lipid extraction, following Bligh and Dyer’s (1959) method alongside Card’s (1973) modifications (Bligh and Dyer 1959; Card 1973).”
Influential
Using algae to treat wastewater from vegetables cooking: screeningScreening algal strains and assessing bacterial communities
Wei, J., Ye, S., Rao, M., Ta, M., Wang, M., & Li, M. (2024) · Desalination and Water Treatment · doi:10.1016/j.dwt.2024.100085 · Cited by 4
Qipengyuania benthica sp. nov. and Qipengyuania profundimaris sp. nov., two novel Erythrobacteraceae members isolated from deep-sea environments
Wang, Y., You, H., Bu, Y., et al. (2024) · INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY · doi:10.1099/ijsem.0.006313 · Cited by 4
Physicochemical composition, centesimal, and phytochemical profile of umbu seed flour (Spondias tuberosa Arruda)
Freitas, E. D. C., Ribeiro, A. B., Miranda, A. D. S., Santana, R. F., Matos, T. B., Gomes, J. R., Santana, J. D. M., & Milagres, M. P. (2024) · Food Science and Technology · doi:10.5327/fst.00242 · Cited by 4
Evaluation of the nutritional potential of farmed Atlantic salmon (Salmo salar) tissues based on the contents of essential fatty acids
Mai, T. T. H., Choi, J., Park, D. H., Park, S.-O., & Kim, H. (2024) · Aquaculture Reports · doi:10.1016/j.aqrep.2024.102119 · Cited by 4
Soybean molasses increases subcutaneous fat deposition while reducing lipid oxidation in the meat of castrated lambs
Pereira-Junior, S. A. G., Costa, R. V., Rodrigues, J. L., et al. (2024) · Journal of Animal Science · doi:10.1093/jas/skae130 · Cited by 4
“Intramuscular fat content was measured as total lipids through cold extraction by Bligh and Dyer (1959).”
Method
Effects of dietary supplementation with inactivated Lactobacillus plantarum on growth performance, haemato-biochemical parameters, liver fatty acids profile and intestinal microbiome of Nile tilapia
Ferro, P., Ribeiro, G. C., Borba, L. E., et al. (2024) · Veterinary Research Communications · doi:10.1007/s11259-024-10425-w · Cited by 4
Influence of Cuttlefish-Ink Extract on Canned Golden Seabream (Sparus aurata) Quality
Martínez, B., Trigo, M., Rodrı́guez, A., et al. (2024) · Foods · doi:10.3390/foods13111685 · Cited by 4
Temporal gene expression during asexual development of the apicomplexan Sarcocystis neurona
Dangoudoubiyam, S., Norris, J. K., Namasivayam, S., et al. (2024) · mSphere · doi:10.1128/msphere.00111-24 · Cited by 4
“A portion of this sample was treated to remove lipids to collect protein samples with and without the presence of lipids (44).”
Background
Bosea rubneri sp. nov. Isolated from Organically Grown Allium cepa
Stoll, D., Grimmler, C., Hetzer, B., et al. (2024) · Current Microbiology · doi:10.1007/s00284-024-03717-6 · Cited by 4
“…analyses of strain ZW T0_25 T were carried out by the Identification Service of the DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) including analyses of respiratory quinones [15, 16], fatty acids [15, 17–21], whole cell sugars [22, 23] and polar lipids [24, 25].”
Postpartum development of metabolic dysfunction-associated steatotic liver disease in a lean mouse model of gestational diabetes mellitus
Hribar, K., Eichhorn, D., Bongiovanni, L., Koster, M. H., Kloosterhuis, N., de Bruin, A., Oosterveer, M. H., Kruit, J. K., & van der Beek, E. M. (2024) · Scientific Reports · doi:10.1038/s41598-024-65239-2 · Cited by 4
“Hepatic lipids were extracted from 15% (w/v) liver homogenates in PBS, according to the method described by Bligh and Dyer 24,25 .”
Method
Lipid Extraction Methods from Wet Microalgal Biomass
Meena, S. S., Mohanty, A., Pandit, N. K., & Mann, S. S. (2024) · Environmental science and engineering · doi:10.1007/978-3-031-52319-9_6 · Cited by 4
Biomarker responses in Danio rerio following an acute exposure (96 h) to e-waste leachate
Rielly, A., Dahms-Verster, S., & Greenfield, R. (2024) · Ecotoxicology · doi:10.1007/s10646-024-02784-6 · Cited by 5
“…at 490 nm. Protein availability was determined using the methodology of Bradford (1976) and lipid availability by following the procedure outlined by Bligh and Dyer (1959); where 500 µL of sample supernatant and 500 µL of chloroform were added to sterile microcentrifuge tubes and vortex thoroughly.”
Method
Nitro fatty acids: A comprehensive review on analytical methods and levels in health and disease
Elshoura, Y., Herz, M., Gad, M. Z., & Hanafi, R. S. (2024) · Analytical Biochemistry · doi:10.1016/j.ab.2024.115624 · Cited by 6
Influence of dietary bitter orange peel powder on growth, body composition, blood parameters, gut morphometry, and thermal tolerance of Nile tilapia (Oreochromis niloticus)
Gressler, L. T., Centenaro, J. R., Braz, P. H., Costa, S. Z. R., Battisti, E. K., Gressler, L. T., Finamor, I., & Sutili, F. J. (2024) · Fish Physiology and Biochemistry · doi:10.1007/s10695-024-01383-w · Cited by 4
“Six fish/ treatment (pool of two fish/tank) were used to analyze whole-body centesimal composition using standard procedures according to AOAC (1995) and Bligh and Dyer (1959).”
Mesorhizobium retamae sp. nov., a novel non-nodulating and non-nitrogen-fixing species isolated from the root nodules of Retama raetam sampled in Tunisia
Gaied, R. B., Sbissi, I., Tarhouni, M., et al. (2024) · INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY · doi:10.1099/ijsem.0.006478 · Cited by 4

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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