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700
Industrial Park Drive
Alabaster, AL 35007
1-800-227-0651 or 205-663-2494
Fax 1-800-229-1004 or 205-663-0756
Orders:
Technical Questions:
General
Information:
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Technical
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| Protonation
of Acidic Lipids |
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Theory
Protonate the phospholipid using acid. Wash the excess
acid and salt from the product.
Equipment
- Separatory funnel (teflon stopcock) or suitable glass
vessel
- Rotary evaporator or nitrogen/argon stream
- pH indicating paper
Reagents
Chloroform
Methanol
HCl
Deionized water
Procedure
- Dissolve the lipid in a volume of chloroform: methanol
(2:1 v/v) such that the lipid con centration is 10-50 mg/ml.
(e.g. 9.0 ml)
- Add deionized water to the solution. (0.2 times the volume
of the chloroform: methanol: e.g. 1.8 ml)
- Shake the two solutions and let the phases separate.
- Adjust the water phase to < pH 3.0 using acid. (pKa
of the phosphate is approximately 3.5.)
- Shake the acidified solution and allow the phases to
separate.
- Isolate the bottom phase. (This phase contains the acidified
phospholipid.)
- Remove the chloroform by rotary evaporation or nitrogen/argon
stream under a hood.
- Dissolve the residue in a suitable organic solvent.
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