title: A novel automated hydrophilic interaction liquid chromatography method using diode-array detector/electrospray ionization tandem mass spectrometry for analysis of sodium risedronate and related degradation products in pharmaceuticals.
abstract: A simple, sensitive and fast hydrophilic interaction liquid chromatography (HILIC) method using ultraviolet diode-array detector (UV-DAD)/electrospray ionization tandem mass spectrometry was developed for the automated high performance liquid chromatography (HPLC) determination of sodium risedronate (SR) and its degradation products in new pharmaceuticals. The chromatographic separations were performed on Ascentis Express HILIC 2.7μm (150mm×2.1mm, i.d.) stainless steel column (fused core). The mobile phase consisted of formate buffer solution (pH 3.4; 0.03M)/acetonitrile 42:58 and 45:55 (v/v) for granules for oral solution and effervescent tablet analysis, respectively, at a flow-rate of 0.2mL/min, setting the wavelength at 262nm. Stability characteristics of SR were evaluated by performing stress test studies. The main degradation product formed under oxidation conditions corresponding to sodium hydrogen (1-hydroxy-2-(1-oxidopyridin-3-yl)-1-phosphonoethyl)phosphonate was characterized by high performance liquid chromatography-electrospray ionization-mass tandem mass spectrometry (HPLC-ESI-MS/MS). The validation parameters such as linearity, sensitivity, accuracy, precision and selectivity were found to be highly satisfactory. Linear responses were observed in standard and in fortified placebo solutions. Intra-day precision (relative standard deviation, RSD) was ≤1.1% for peak area and ≤0.2% for retention times (tR) without significant differences between intra- and inter-day data. Recovery studies showed good results for all the examined compounds (from 98.7 to 101.0%) with RSD ranging from 0.6 to 0.7%. The limits of detection (LOD) and quantitation (LOQ) were 1 and 3ng/mL, respectively. The high stability of standard and sample solutions at room temperature means an undoubted advantage of the method allowing the simultaneous preparation of many samples and consecutive chromatographic analyses by using an autosampler. The developed stability indicating method is suitable for the quality control of SR in new and commercial pharmaceutical formulations.
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123 Trento, TN, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Bertolini
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123 Trento, TN, Italy.
authors.forename: Lorenza
authors.initials: L
authors.lastname: Vicentini
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123 Trento, TN, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Boschetti
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123 Trento, TN, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Andreatta
authors.affiliation: Department of Pharmacy & Biotechnology, Alma Mater Studiorum - Università di Bologna, Via Belmeloro 6, 40126 Bologna, Italy. Electronic address: rita.gatti2@unibo.it.
authors.forename: Rita
authors.initials: R
authors.lastname: Gatti
mesh_terms: D050071:Bone Density Conservation Agents / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D012968:Etidronic Acid / Q000031:analogs & derivatives* / Q000032:analysis; D057927:Hydrophobic and Hydrophilic Interactions; D057230:Limit of Detection; D011786:Quality Control; D015203:Reproducibility of Results; D000068296:Risedronic Acid; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013607:Tablets; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D050071:Bone Density Conservation Agents; D013607:Tablets; D000068296:Risedronic Acid; D012968:Etidronic Acid
keywords: Degradation products; HILIC fused core technology; HPLC-ESI-MS; Pharmaceutical formulations; Sodium risedronate; Stability indicating