title: Single column comprehensive analysis of pharmaceutical preparations using dual-injection mixed-mode (ion-exchange and reversed-phase) and hydrophilic interaction liquid chromatography.
abstract: The comprehensive separation and detection of hydrophobic and hydrophilic active pharmaceutical ingredients (APIs), their counter-ions (organic, inorganic) and excipients, using a single mixed-mode chromatographic column, and a dual injection approach is presented. Using a mixed-mode Thermo Fisher Acclaim Trinity P1 column, APIs, their counter-ions and possible degradants were first separated using a combination of anion-exchange, cation-exchange and hydrophobic interactions, using a mobile phase consisting of a dual organic modifier/salt concentration gradient. A complementary method was also developed using the same column for the separation of hydrophilic bulk excipients, using hydrophilic interaction liquid chromatography (HILIC) under high organic solvent mobile phase conditions. These two methods were then combined within a single gradient run using dual sample injection, with the first injection at the start of the applied gradient (mixed-mode retention of solutes), followed by a second sample injection at the end of the gradient (HILIC retention of solutes). Detection using both ultraviolet absorbance and refractive index enabled the sensitive detection of APIs and UV-absorbing counter-ions, together with quantitative determination of bulk excipients. The developed approach was applied successfully to the analysis of a dry powder inhalers (Flixotide(®), Spiriva(®)), enabling comprehensive quantification of all APIs and excipients in the sample.
authors.affiliation: Pfizer Analytical Research Centre, Australian Centre for Research on Separation Science, School of Chemistry, University of Tasmania, Private Bag 75, Hobart, Tasmania 7001, Australia.
authors.forename: Artaches A
authors.initials: AA
authors.lastname: Kazarian
authors.forename: Mark R
authors.initials: MR
authors.lastname: Taylor
authors.forename: Paul R
authors.initials: PR
authors.lastname: Haddad
authors.forename: Pavel N
authors.initials: PN
authors.lastname: Nesterenko
authors.forename: Brett
authors.initials: B
authors.lastname: Paull
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D004364:Pharmaceutical Preparations / Q000032:analysis*
chemical_list: D004364:Pharmaceutical Preparations
keywords: Active pharmaceutical ingredients; Excipients; HILIC; Ion-exchange; Mixed-mode chromatography