Determination of fexofenadine in human plasma using 96-well solid phase extraction and HPLC with tandem mass spectrometric detection

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 35; no. 4; pp. 837 - 846
• Main Authors: Fu, I., Woolf, E.J., Matuszewski, B.K.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 29.06.2004; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Antihistamine; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Fexofenadine; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Humans; Mass spectrometry; Mass Spectrometry - methods; Medical sciences; P-glycoprotein; Pharmacology. Drug treatments; Plasma; Terfenadine - analogs & derivatives; Terfenadine - blood; Terfenadine - chemistry; P-glycoprotein; Plasma; Fexofenadine; Antihistamine; Mass spectrometry; HPLC; Human; Solid phase extraction; Biological fluid; Ammonium; Single dose; Ion pair; HPLC chromatography; Antihistaminic; Chemical ionization; Blood plasma; Mass spectrometry MS/MS; Antagonist; H1 Histamine receptor; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2004.02.016

A fast and sensitive HPLC–MS/MS method, utilizing atmospheric pressure chemical ionization, for the determination of fexofenadine in human plasma is described. A deuterated analog, d6-fexofenadine is used as the internal standard (IS). Plasma samples are prepared using 96-well solid phase extraction with plates containing Waters Oasis HLB sorbent. The analytes are chromatographed on a Restek Ultra IBD column (3.2mm×50mm, 3μm) using a mobile phase consisting of a mixture of 90% acetonitrile and 10% 10mM ammonium acetate buffer and 0.1% formic acid. Quantitation of the analyte is based on the response from the multiple reaction monitoring of the precursor to product ion pairs for fexofenadine (m/z 502→466) and d6-fexofenadine (m/z 508→472). The assay has been validated over the concentration range of 1–200ng/ml based on the analysis of 0.5ml aliquots of plasma. Within-day assay accuracy was between 97 and 102% of nominal, while within-day precision was better than 3.5% CV at all points on the standard curve. Analyte extraction recovery was better than 70% over the range of the standard curve. The method was found to be suitable for the analysis of human plasma samples obtained 24h following the administration of a single 60mg dose of fexofenadine.