What are the methods for extracting lipids from tissues in lipidomics research?
In lipid metabolomics research, common methods for extracting lipids from tissues are as follows:
1. Sample Preparation：
- Collect tissue samples: Ensure the samples are fresh and freeze them quickly to prevent lipid oxidation.
- Weighing: Accurately record the weight of the sample for subsequent calculation of lipid concentration.
2. Homogenization：
Use a homogenizer (such as a bead mill, ultrasonic disruptor, etc.) under cooling conditions to homogenize tissue samples and release intracellular lipids.
3. Lipid Extraction：
The Bligh and Dyer method is suitable for small sample volumes.
- Add a mixed solvent of methanol and chloroform to the sample (usually in a 2:1 ratio).
- After thorough mixing, add chloroform and water to achieve a final ratio of 1:1:0.9 (methanol: chloroform: water).
- Centrifuge and collect the lower chloroform layer containing lipids.
Folch method: Suitable for larger sample sizes.
- Use a mixed solvent of chloroform and methanol (ratio 2:1).
- Add a certain amount of 0.9% saline.
- After centrifugation, also collect the lower chloroform layer.
4. Washing：
It is possible to wash by adding a certain proportion of saline solution and centrifuging again to remove non-lipid impurities from the upper aqueous phase.
5. Lipid Concentration：
Use a rotary evaporator or nitrogen blower to evaporate the solvent at low temperature, leaving pure lipids.
6. Solvent Evaporation：
Evaporate the solvent under nitrogen or in a vacuum dryer, leaving pure lipids.
7. Lipid Resuspension：
Resuspend the extracted lipids in an appropriate solvent (such as chloroform) for subsequent analysis.
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