title: Hydrophilic interaction liquid chromatography in analysis of granisetron HCl and its related substances. Retention mechanisms and method development.
abstract: In this paper separation of granisetron and its two related substances in HILIC mode is presented. Separation was done on silica column derivatized with sulfoalkylbetaine groups (ZIC-HILIC). Firstly, retention mechanisms were assessed whereby retention factors of substances were followed in wide range of acetonitrile content (80-97%), at constant concentration of aqueous buffer (10mM) as well as at constant pH value of 3.0. Further, in order to developed optimal HILIC method, Design of Experiments (DoE) methodology was applied. For optimization full factorial design 3(2) was employed. Influence of acetonitrile content and ammonium acetate concentration were investigated while pH of the water phase was kept at 3.3. Adequacy of obtained mathematical models was confirmed by ANOVA. Optimization goals (α>1.15 and minimal run time) were accomplished with 94.7% of acetonitrile in mobile phase and 70 mM of ammonium acetate in water phase. Optimal point was in the middle of defined Design Space. In the next phase, robustness was experimetally tested by Rechtschaffen design. The investigated factors and their levels were: acetonitrile content (±1%), ammonium acetate molarity in water phase (±2 mM), pH value of water phase (±0.2) and column temperature (±4 °C). The validation scope included selectivity, linearity, accuracy and precision as well as determination of limit of detection (LOD) and limit of quantification (LOQ) for the related substances. Additionally, the validation acceptance criteria were met in all cases. Finally, the proposed method could be successfully utilized for estimation of granisetron HCl and its related substances in tablets and parenteral dosage forms, as well as for monitoring degradation under various stress conditions.
authors.affiliation: Military Medical Academy, Sector for Pharmacy, Department of Drug Control and Examination, Crnotravska 17, Belgrade, Serbia.
authors.forename: Jelena
authors.initials: J
authors.lastname: Maksić
authors.affiliation: University of Belgrade-Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, 11000 Belgrade, Serbia.
authors.forename: Anja
authors.initials: A
authors.lastname: Tumpa
authors.affiliation: University of Belgrade-Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, 11000 Belgrade, Serbia.
authors.forename: Ana
authors.initials: A
authors.lastname: Stajić
authors.affiliation: University of Belgrade-Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, 11000 Belgrade, Serbia.
authors.forename: Marko
authors.initials: M
authors.lastname: Jovanović
authors.affiliation: University of Belgrade-Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, 11000 Belgrade, Serbia.
authors.forename: Tijana
authors.initials: T
authors.lastname: Rakić
authors.affiliation: University of Belgrade-Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, 11000 Belgrade, Serbia. Electronic address: jancic.stojanovic@pharmacy.bg.ac.rs.
authors.forename: Biljana
authors.initials: B
authors.lastname: Jančić-Stojanović
mesh_terms: D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D004304:Dosage Forms; D017829:Granisetron / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D057230:Limit of Detection; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry; D013607:Tablets / Q000737:chemistry; D013696:Temperature; D014867:Water / Q000737:chemistry
chemical_list: D000085:Acetates; D000097:Acetonitriles; D004304:Dosage Forms; D017829:Granisetron; D012822:Silicon Dioxide; D013607:Tablets; D014867:Water; C018824:ammonium acetate; C032159:acetonitrile
keywords: Granisetron; Hydrophilic interaction liquid chromatography; Optimization; Releted substances; Retention mechanisms