title: High performance liquid chromatography analysis of 100-nm liposomal nanoparticles using polymer-coated, silica monolithic columns with aqueous mobile phase.
abstract: Recently, nanoparticles have garnered considerable attention, and the demand for a rapid and simple method for their analysis has increased accordingly. The bimodal pores (few μm- and few tens nm-sized pores) of monolithic columns were thought to be suitable for the separation of nanoparticles and small molecules; however, the residual silanol groups on the column surface resulted in the strong adsorption of liposomes and hindered their analysis. To overcome this problem, we modified the surface of the silica monolith via a two-step process and developed three silica monolithic columns coated with three different polymers: glycidyl methacrylate (GMA), 2-hydroxyethyl methacrylate (HEMA), and N-vinylpyrrolidone (VP). These were used for the analysis of 100-nm liposomal nanoparticles. Since 15% polymer coating prevented the nanoparticle adsorption, liposomes (AmBisome®) and pegylated liposomes (DOXIL®) were eluted rapidly (within 1min) using these columns, without using organic solvents in the mobile phase. Molecular leaching from the liposomes, as well as protein adsorption to the liposomes (corona formation) could be evaluated using the polymer-coated columns, thus illustrating their utility in the rapid and simple analysis of 100-nm liposomal nanoparticles.
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Itoh
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba, 278-8510, Japan.
authors.forename: Arato
authors.initials: A
authors.lastname: Kimoto
authors.affiliation: Formulation Research, Pharmaceutical Science and Technology, Core Function Unit, Medicine Development Center, Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki, 300-2635, Japan.
authors.forename: Eiichi
authors.initials: E
authors.lastname: Yamamoto
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba, 278-8510, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Higashi
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Santa
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Funatsu
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. Electronic address: masaru-kato@umin.ac.jp.
authors.forename: Masaru
authors.initials: M
authors.lastname: Kato
mesh_terms: D000327:Adsorption; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004852:Epoxy Compounds / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry* / Q000302:isolation & purification; D008689:Methacrylates / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D011760:Pyrrolidinones / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D004852:Epoxy Compounds; D008081:Liposomes; D008689:Methacrylates; D011506:Proteins; D011760:Pyrrolidinones; C005044:hydroxyethyl methacrylate; D012822:Silicon Dioxide; C042670:N-vinyl-2-pyrrolidinone; C007870:glycidyl methacrylate
keywords: 100nm liposomal nanoparticle; Aqueous solution; Polymer coat; Silica monolith