Lesson 5.2. Isolation/Extraction of Lipids

Homogenize

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

1 = ?

50 grams of the samples

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

2 = ?

Chloroform:methanol (2:1) or Hexane:methanol (2:1)

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

3 = ?

5 minutes

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

4 = ?

lipid extract

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

5 = ?

anhydrous Na2SO4

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

6 = ?

extract

Extraction of Lipids (Process):

1. ___________________ the samples

2. Weigh __________________________

3. Add 50 mL of extracting solvent, _____________________________________

4. Mix thoroughly and let the mixture stand for ___________________.

5. Collect the _______________ in a beaker by decantation.

6. Add _________________to remove residual water.

7. Decant the _______________ to another container to separate the Na2SO4.

7 = ?

oxidation & peroxidation

Susceptibility of lipids to ____________________________

subdivided

Conditions that must be observed during extraction:

a. Tissues must be ___________ under conditions that will not bring about degradation of lipids

b. Solvents to be use must _________________________________________

c. Tissue must be washed completely free of lipid by ____________________ with lipid-free solvent

a = ?

penetrate the tissue & break protein-lipid bond

Conditions that must be observed during extraction:

a. Tissues must be ___________ under conditions that will not bring about degradation of lipids

b. Solvents to be use must _________________________________________

c. Tissue must be washed completely free of lipid by ____________________ with lipid-free solvent

b = ?

repeated treatment

Conditions that must be observed during extraction:

a. Tissues must be ___________ under conditions that will not bring about degradation of lipids

b. Solvents to be use must _________________________________________

c. Tissue must be washed completely free of lipid by ____________________ with lipid-free solvent

c = ?

Homogenization

____________________ by triturating with sand or using an osterizer breaks the integrity of the cell’s membranes

simple washing

Non-lipid components can be removed by _________________ while lipids remain dissolved in the organic solvent

below 4 degrees

To prevent oxidation & peroxidation, tissues can be cooled to ________________ immediately following excision

Triacylglycerols

extracted with acetone or diethyl ether

lecithins

Phosphoglycerides that are soluble in ether, CHCl3, C6H6, hot alcohol, CS2, insoluble in acetone

cephalins

Phosphoglycerides that are soluble in ether, CHCl3, C6H6, CS2, insoluble in acetone, ethyl & methyl alcohols

Glycolipids

insoluble in ether; more soluble in acetone than phospholipids, soluble in hot alcohol, CHCl3, C6H6

Cholesterol

soluble in ether, CHCl3, C6H6, & hot alcohol and easily crystallize from such solutions