title: HPLC analysis as a tool for assessing targeted liposome composition.
abstract: Functionalized phospholipids are indispensable materials for the design of targeted liposomes. Control over the quality and quantity of phospholipids is thereby key in the successful development and manufacture of such formulations. This was also the case for a complex liposomal preparation composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), Cholesterol (CHO), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000). To this end, an RP-HPLC method was developed. Detection was done via evaporative light scattering (ELS) for liposomal components. The method was validated for linearity, precision, accuracy, sensitivity and robustness. The liposomal compounds had a non-linear quadratic response in the concentration range of 0.012-0.42 mg/ml with a correlation coefficient greater than 0.99 with an accuracy of method confirmed 95-105% of the theoretical concentration. Furthermore, degradation products from the liposomal formulation could be identified. The presented method was successfully implemented as a control tool during the preparation of functionalized liposomes. It underlined the benefit of HPLC analysis of phospholipids during liposome preparation as an easy and rapid control method for the functionalized lipid at each preparation step as well as for the quantification of all components.
authors.affiliation: Merck Serono, Chemical & Pharmaceutical Development, Frankfurter Straße 250, 64293 Darmstadt, Germany. Electronic address: Mira.Oswald@external.merckgroup.com.
authors.forename: Mira
authors.initials: M
authors.lastname: Oswald
authors.affiliation: Merck & Cie, MM-PPR-FPL Lipids 1, Im Laternenacker 5, 8200 Schaffhausen, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Platscher
authors.affiliation: Merck Serono, Chemical & Pharmaceutical Development, Frankfurter Straße 250, 64293 Darmstadt, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Geissler
authors.affiliation: University of Regensburg, Department for Pharmaceutical Technology, Universitätsstraße 31, 94030 Regensburg, Germany. Electronic address: achim.goepferich@chemie.uni-regensburg.de.
authors.forename: Achim
authors.initials: A
authors.lastname: Goepferich
mesh_terms: D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004337:Drug Carriers / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D010743:Phospholipids / Q000032:analysis* / Q000737:chemistry*
chemical_list: D004337:Drug Carriers; D006020:Glycopeptides; D008081:Liposomes; D010743:Phospholipids
keywords: DSPE-PEG(2000); ELSD; HPLC; Liposomes; Phospholipid analysis