Liver lipids were extracted, according to a modified Folch method (12 (link)). Frozen liver tissue (100 mg) was thoroughly homogenized with 1 ml of a 2:1 ratio choloroform:methanol mixture (Sigma-Aldrich, Dorset, UK). Deionized water (200 μl) was then added to the mix, vortexed, and shaken for 10 min. Samples were centrifuged for 10 min at 16,100 g to generate a distinct organic and aqueous phase. The lower organic phase (500 μl) was collected into a preweighed glass tube and dried using a rotary evaporator for 2 h at 37°C, and the remaining lipid content was weighed.
Choloroform methanol mixture
Manufactured by Merck Group
Sourced in United Kingdom
Chloroform:methanol mixture is a laboratory reagent commonly used for the extraction and purification of lipids and other organic compounds. It is a binary solvent system composed of chloroform and methanol in a specific ratio. This mixture effectively solubilizes a wide range of polar and non-polar compounds, making it a versatile tool for analytical and preparative applications in biochemistry, molecular biology, and organic chemistry laboratories.
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2 protocols using choloroform methanol mixture
1
Extraction and Quantification of Liver Lipids
2
Hepatic Lipid Quantification by Folch Method
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Accumulation of hepatic lipid was determined using a modified Folch method. In brief, 25 mg of frozen liver was homogenized in 1.2 ml of a 2:1 ratio choloroform:methanol mixture (Sigma-Aldrich, Dorset, UK). Deionized water (240 μl) was then added to the mix, vortexed thoroughly, and samples centrifuged for 10 min at 16,100 × g to generate a distinct organic and aqueous phase. The lower organic phase (500 μl) was collected into a pre-weighed glass tube and dried under a nitrogen stream with the lipid content weighed and normalised to total liver weight.
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