Determination of the quaternary ammonium compound trospium in human plasma by LC–MS/MS: Application to a pharmacokinetic study

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2010-04, Vol.878 (13), p.981-986
• Main Authors: Hotha, Kishore Kumar, Bharathi, D. Vijaya, Kumar, Sanagapati Sirish, Reddy, Y. Narasimha, Chatki, Pankaj K., Ravindranath, L.K., Jayaveera, K.N.
• Format: Article
• Language: English
• Subjects: Ammonium compounds; Analysis; Analytical, structural and metabolic biochemistry; Battery; Benzilates; Biological and medical sciences; Cations; Chromatography, Liquid - methods; Elution; Extraction; Fundamental and applied biological sciences. Psychology; General pharmacology; Guidelines; Human; Human plasma; LC–MS/MS; Limit of Detection; Medical sciences; Nortropanes - blood; Nortropanes - pharmacokinetics; Pharmacokinetics; Pharmacology. Drug treatments; Quaternary Ammonium Compounds - blood; Quaternary Ammonium Compounds - pharmacokinetics; Quaternary compound; Run time (computers); Tandem Mass Spectrometry - methods; Trospium
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.02.028

A highly sensitive, specific and evaporation free SPE extraction, LC–MS/MS method has been developed for the estimation of trospium in human plasma using trospium-d8 as an internal standard (IS). The analyte was separated using isocratic mobile phase on reverse phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M +] cations, m/ z 392–164 for trospium and m/ z 400–172 for the IS. The total run time was 3.50 min and the elution of trospium and trospium-d8 (IS) occurred at 2.8 min. The developed method was validated in human plasma with a lower limit of quantification of 0.05 ng/mL. A linear response function was established for the range of concentrations 0.05–10 ng/mL ( r > 0.998) for trospium in human plasma. The intra- and inter-day precision values for trospium met the acceptance as per FDA guidelines. Trospium was stable in the battery of stability studies viz., bench-top, auto-sampler, dry extracts and freeze/thaw cycles. The developed assay method was applied to an oral pharmacokinetic study in humans.