300290 Isolation of Different Classes of Lipids

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Isolation of Different Classes of Lipids

Summary

Type: SPE
Application #: 300290
Catalog#: 730033 - SPE Cartridges, Chromabond, NH2, 3ml, PP, 500mg, 50/pk
Detection:
Conditions: Column conditioning:
10 ml hexane

Sample application:
Slowly force or aspirate 5 ml of the lipid-containing chloroform extract
through the 1st column

Elution 1: (column 1)
neutral lipids: 4 ml chloroform/2-propanol (2:1, v/v)
fatty acids: 4 ml diethyl ether/acetic acid (98:2, v/v)
phospholipids: 4 ml methanol
The fraction of the neutral lipids is evaporated, redissolved in hexane and
applied to the 2nd column which was conditioned with hexane too.

Elution 2: (column 2)
cholesteryl esters: 4 ml hexane
Since during elution of the triglycerides cholesterol may be eluted too, a
third column is placed under the second column during the next elution step.

Elution 3: (through joined columns 2 and 3)
triglycerides: 4 ml hexane/diethyl ether/methylenechloride (89:1:10, v/v/v)

Elution 4: (through separated columns 2 and 3)
cholesterol: 4 ml each of hexane/ethyl acetate (95:5, v/v)

Elution 5: (column 2)
diglycerides: 4 ml hexane/ethyl acetate (85:15, v/v)
monoglycerides: 4 ml chloroform/methanol (2:1, v/v)
Authors: Macherey-Nagel
Source: LC Department


Details


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