Direct lipid quantitation of cationic liposomes by reversed-phase HPLC in lipoplex preparation process.
The proposed reversed-phase high-performance liquid chromatography method with ultraviolet detection provides a simple and rapid procedure to separate and quantitate lipids from cationic liposomes used in gene transfer. We describe experimental conditions which do not require lipid extraction from liposomes prior to sample analysis. Evaluation of the method reported here showed suitable lipid separation capacity and quantitation accuracy from cationic liposomes composed of either the pentammonio lipid pcTG90 and dioleoyl phosphatidylethanolamine, or 1, 2-dioleoyl-3-trimethylamonium propane and cholesterol. Detection limits were in the range of 0.5-1 microg depending on the lipid. This quantitative method has proven useful in lipoplex formulation processing development and its application may be extended to a wide range of lipid-based gene and drug delivery systems.
Meyer
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O
Vectorology and Biochemistry, TRANSGENE S.A., Strasbourg, Cedex, France. olivier.meyer@transgene.fr
Roch
O
O
Elmlinger
D
D
Kolbe
H V
HV
D002412:Cations / Q000032:analysis; D002784:Cholesterol / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005229:Fatty Acids, Monounsaturated / Q000032:analysis / Q000302:isolation & purification; D020404:Glycerophospholipids / Q000032:analysis / Q000302:isolation & purification; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification; D008081:Liposomes / Q000032:analysis*; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds / Q000032:analysis / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D002412:Cations; D005229:Fatty Acids, Monounsaturated; D020404:Glycerophospholipids; D008055:Lipids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D002784:Cholesterol; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane