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
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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).
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Conflict of interest
The authors declare no potential conflict of interest.
Brynne N, Forslund C, Halle B, Gustafsson LL, Bertilsson L (1999a) Ketoconazole inhibits the metabolism of tolterodine in subjects with deficient CYP2D6 activity. Br J Clin Pharmacol 48:564–572CrossRefPubMedPubMedCentralGoogle Scholar
Brynne N, Svanström C, Aberg-Wistedt A, Hallén B, Bertilsson L (1999b) Fluoxetine inhibits the metabolism of tolterodine-pharmacokinetic implications and proposed clinical relevance. Br J Clin Pharmacol 48:553–563CrossRefPubMedPubMedCentralGoogle Scholar
Byeon JY, Kim YH, Na HS, Jang JH, Kim SH, Lee YJ, Bae JW, Kim IS, Jang CG, Chung MW, Lee SY (2015) Effects of the CYP2D6*10 allele on the pharmacokinetics of atomoxetine and its metabolites. Arch Pharm Res 38:2083–2091CrossRefPubMedGoogle Scholar
Chaudhry SR, Muhammad S, Eidens M, Klemm M, Khan D, Efferth T, Weisshaar MP (2014) Pharmacogenetic prediction of individual variability in drug response based on CYP2D6, CYP2C9 and CYP2C19 genetic polymorphisms. Curr Drug Metab 15:711–718CrossRefPubMedGoogle Scholar
Chen F, Hu ZY, Laizure SC, Hudson JQ (2017) Simultaneous assay of multiple antibiotics in human plasma by LC-MS/MS: importance of optimizing formic acid concentration. Bioanalysis 9:469–483CrossRefPubMedGoogle Scholar
Halle B, Brynne N, Svanstro C (1999) Fluoxetine inhibits the metabolism of tolterodine—pharmacokinetic. Br J Clin Pharmacol 48:553–563Google Scholar
Heaton JC, Russell JJ, Underwood T, Boughtflower R, McCalley DV (2014) Comparison of peak shape in hydrophilic interaction chromatography using acidic salt buffers and simple acid solutions. J Chromatogr A 1347:39–48CrossRefPubMedGoogle Scholar
Kim SH, Kim DH, Byeon JY, Kim YH, Kim DH, Lim HJ, Lee CM, Whang SS, Choi CI, Bae JW, Lee YJ, Jang CG, Lee SY (2017) Effects of CYP2C9 genetic polymorphisms on the pharmacokinetics of celecoxib and its carboxylic acid metabolite. Arch Pharm Res 40:382–390CrossRefPubMedGoogle Scholar
Lee YJ, Byeon JY, Kim YH, Kim SH, Choi CI, Bae JW, Sohn UD, Jang CG, Lee J, Lee SY (2015) Effects of CYP2C9*1/*3 genotype on the pharmacokinetics of flurbiprofen in Korean subjects. Arch Pharm Res 38:1232–1237CrossRefPubMedGoogle Scholar
Lee HJ, Kim YH, Kim SH, Lee CM, Yang AY, Jang CG, Lee SY, Bae JW, Choi CI (2016) Effects of CYP2C9 genetic polymorphisms on the pharmacokinetics of zafirlukast. Arch Pharm Res 39:1013–1019CrossRefPubMedGoogle Scholar
Macek J, Ptácek P, Klíma J (2009) Determination of tolterodine and its 5-hydroxymethyl metabolite in human plasma by hydrophilic interaction liquid chromatography—tandem mass spectrometry. J Chromatogr B 877:968–974CrossRefGoogle Scholar
Palmér L, Andersson L, Andersson T, Stenberg U (1997) Determination of tolterodine and the 5-hydroxymethyl metabolite in plasma, serum and urine using gas chromatography-mass spectrometry. J Pharm Biomed Anal 16:155–165CrossRefPubMedGoogle Scholar
Postlind H, Danielson A, Lindgren A, Andersson SH (1998) Tolterodine, a new muscarinic receptor antagonist, is metabolized by cytochromes P450 2D6 and 3A in human liver microsomes. Drug Metab Dispos 26:289–293PubMedGoogle Scholar
Rentzhog L, Stanton SL, Cardozo L, Nelson E, Fall M, Abrams P (1998) Efficacy and safety of tolterodine in patients with detrusor instability: a dose-ranging study. Br J Urol 81:42–48CrossRefPubMedGoogle Scholar
Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, Shah VP, Skelly JP, Swann PG, Weiner R (2007) Quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. Pharm Res 24:1962–1973CrossRefPubMedGoogle Scholar
Yanamandra R, Vadla CS, Puppala U, Patro B, Murthy YL, Ramaiah PA (2012) A new rapid and sensitive stability-indicating UPLC assay method for tolterodine tartrate: application in pharmaceuticals, human plasma and urine samples. Sci Pharm 80:101–114CrossRefPubMedGoogle Scholar
Zhang B, Zhang Z, Tian Y, Xu F (2005) High-performance liquid chromatography-electrospray ionization mass spectrometric determination of tolterodine tartrate in human plasma. J Chromatogr B 824:92–98CrossRefGoogle Scholar