Archives of Pharmacal Research

, Volume 40, Issue 11, pp 1287–1295 | Cite as

Simultaneous determination of tolterodine and its two metabolites, 5-hydroxymethyltolterodine and N-dealkyltolterodine in human plasma using LC–MS/MS and its application to a pharmacokinetic study

  • Young-Hoon Kim
  • Ji-Yeong Byeon
  • Se-Hyung Kim
  • Choong-Min Lee
  • Eui Hyun Jung
  • Won Ki Chae
  • Choon-Gon Jang
  • Seok-Yong Lee
  • Yun Jeong Lee
Research Article


Tolterodine is a nonselective muscarinic antagonist that is indicated for the overactive urinary bladder and other urinary difficulties. We developed and validated a simple, rapid and sensitive high-performance liquid chromatography analytical method utilizing tandem mass spectrometry (LC–MS/MS) for the quantitation of tolterodine and its major metabolites, 5-hydroxymethyltolterodine (5-HMT) and N-dealkyltolterodine (NDT), in human plasma. After liquid–liquid extraction with methyl t-butyl ether, chromatographic separation of the three analytes was achieved using a reversed-phase Luna Phenyl-hexyl column (100 × 2.0 mm, 3 μm particles) with a mobile phase of 10 mM ammonium formate buffer (pH 3.5)-methanol (10:90, v/v) and quantified by MS/MS detection in electrospray ionization (ESI) positive ion mode. The retention time of tolterodine, 5-HMT, NDT, and internal standard (IS) were 1.4, 1.24, 1.33, and 1.26 min, respectively. The calibration curves were linear over a range of 0.025–10 ng/ml for tolterodine and 5-HMT, and 0.05–10 ng/ml for NDT. The lower limit of quantifications using 200 μl of human plasma was 0.025 ng/ml for tolterodine and 5-HMT, and 0.05 ng/ml for NDT. The mean accuracy and precision for intra- and inter-run validation of tolterodine, 5-HMT, and NDT were all within acceptable limits. These results showed that a simple, rapid and sensitive LC–MS/MS method for the quantification of tolterodine and its major metabolites in human plasma was developed. This method was successfully applied to a pharmacokinetic study in humans.


Tolterodine Hydroxymethyltolterodine N-dealkyltolterodine LC–MS/MS Human plasma 



This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning (NRF-2016R1A2B4007381).

Compliance with ethical standards

Conflict of interest

The authors declare no potential conflict of interest.


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Copyright information

© The Pharmaceutical Society of Korea 2017

Authors and Affiliations

  1. 1.School of PharmacySungkyunkwan UniversitySuwonSouth Korea
  2. 2.College of PharmacyDankook UniversityCheonanSouth Korea

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