title: D-Optimal mixture design optimization of an HPLC method for simultaneous determination of commonly used antihistaminic parent molecules and their active metabolites in human serum and urine.
abstract: This study describes a specific, precise, sensitive and accurate method for simultaneous determination of hydroxyzine, loratadine, terfenadine, rupatadine and their main active metabolites cetirizine, desloratadine and fexofenadine, in serum and urine using meclizine as an internal standard. Solid-phase extraction method for sample clean-up and preconcentration of analytes was carried out using Phenomenex Strata-X-C and Strata X polymeric cartridges. Chromatographic analysis was performed on a Phenomenex cyano (150 × 4.6 mm i.d., 5 μm) analytical column. A D-optimal mixture design methodology was used to evaluate the effect of changes in mobile phase compositions on dependent variables and optimization of the response of interest. The mixture design experiments were performed and results were analyzed. The region of ideal mobile phase composition consisting of acetonitrile-methanol-ammonium acetate buffer (40 mm; pH 3.8 adjusted with acetic acid): 18:36:46% v/v/v was identified by a graphical optimization technique using an overlay plot. While using this optimized condition all analytes were baseline resolved in <10 min. Solvent mixtures were delivered at 1.5 mL/min flow rate and analytes peaks were detected at 222 nm. The proposed bioanalytical method was validated according to US Food and Drug Administration guidelines. The proposed method was sensitive with detection limits of 0.06-0.15 μg/mL in serum and urine samples. Relative standard deviation for inter- and intra-day precision data was found to be <7%. The proposed method may find application in the determination of selected antihistaminic drugs in biological fluids.
authors.affiliation: Department of Pharmacy, Faculty of Engineering and Technology, Annamalai University, Annamalai Nagar, Tamil Nadu, India.
authors.forename: Selvakumar
authors.initials: S
authors.lastname: Kanthiah
authors.affiliation: Department of Pharmacy, Faculty of Engineering and Technology, Annamalai University, Annamalai Nagar, Tamil Nadu, India.
authors.forename: Valliappan
authors.identifier: http://orcid.org/0000-0003-0276-439X
authors.initials: V
authors.lastname: Kannappan
mesh_terms: D018926:Anti-Allergic Agents / Q000097:blood* / Q000378:metabolism / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003533:Cyproheptadine / Q000031:analogs & derivatives / Q000097:blood / Q000378:metabolism / Q000652:urine; D006634:Histamine H1 Antagonists / Q000097:blood* / Q000378:metabolism / Q000652:urine*; D006801:Humans; D006919:Hydroxyzine / Q000097:blood / Q000378:metabolism / Q000652:urine; D057230:Limit of Detection; D017336:Loratadine / Q000097:blood / Q000378:metabolism / Q000652:urine; D052616:Solid Phase Extraction / Q000379:methods; D016593:Terfenadine / Q000097:blood / Q000378:metabolism / Q000652:urine
chemical_list: D018926:Anti-Allergic Agents; D006634:Histamine H1 Antagonists; C103639:rupatadine; D003533:Cyproheptadine; D006919:Hydroxyzine; D017336:Loratadine; D016593:Terfenadine
keywords: D-optimal mixture design methodology; HPLC; hydroxyzine; loratadine; rupatadine; solid-phase extraction; terfenadine