Lipids

> back to Suggested SPE Protocols

Matrix

Tissue

Analytes

Fatty Acids, Phospholipids, Cholesteryl Ester, Triglycerides, Cholesterol, Diglycerides, Monoglycerides

SPE Cartridge

SiliaPrep Amine (WAX)
500 mg / 3 mL
SPE-R52030B-03P

Sample Pretreatment before Loading

Evaporate extract to dryness under Nitrogen and dissolve in 0.5 mL of Chloroform.

Conditionning

No conditionning required

Equilibration

2 x 2 mL of Hexane

Loading

Pretreated sample

Elution

- Elution of Neutral Lipids: 4 mL of Chloroform / 2-Propanol (2:1).
- Elution of Fatty Acids: 4 mL of 2 % Acetic Acid in Diethyl Ether.
- Elution of Phospholipids: 4 mL of Methanol.

With Neutral Lipids fraction: Evaporation under Nitrogen, reconstitution with 0.2 mL of Hexane, load on a second SPE cartridge (same equilibration step).
- Elution of Cholesteryl Ester: 4 mL of Hexane.

Connect a third SPE cartridge below the second one (same equilibration step).
- Elution of Triglycerides: 6 mL of Diethyl Ether / Methylene Chloride / Hexane (1:10:89), through both SPE cartridges.
- Elution of Cholesterol: 12 mL of 5 % Ethyl Acetate in Hexane, through both SPE cartridges.

On the second SPE cartridge only now.
- Elution of Diglycerides: 4 mL of 15 % Ethyl Acetate in Hexane.
- Elution of Monoglycerides: 4 mL of Chloroform / Methanol (2:1).