Transl Clin Pharmacol.  2017 Mar;25(1):21-27. 10.12793/tcp.2017.25.1.21.

Validated UPLC-MS/MS method for the determination of tadalafil in human plasma and its application to a pharmacokinetic study

• Kim KY
• Nam M
• Kwon HJ
• Kim KH
• Kang SH
• Kim SI
• Kim CW
• Cho SH

• KMID: 2411415
• DOI: http://doi.org/10.12793/tcp.2017.25.1.21

Abstract

A simple, rapid, and reliable UPLC-MS/MS method was developed and validated for the determination of tadalafil in human plasma. The plasma samples were deproteinized with acetonitrile. Chromatographic separation was performed on a Shiseido C18 (100 × 2.1 mm, 2.7 µm) column with isocratic elution using 2.0 mM ammonium acetate and acetonitrile (55:45, v/v) with 0.1% formic acid at a flow rate of 0.7 mL/min. The total run time was 1 min per sample. The quantitative analysis was performed using multiple reaction monitoring at transition of m/z 390.4 → 268.3 for tadalafil and m/z 475.3 → 283.3 for sildenafil as an internal standard. The method was fully validated over a concentration range of 5-1,000 ng/mL with a lower quantification limit of 5 ng/mL. Intra- and inter-day precision (relative standard deviation, %RSD) were within 8.4% and accuracy (relative error, %RE) was lower than -3.2%. The developed and validated method was successfully applied to a pharmacokinetic study of tadalafil (20 mg) in Korean healthy male subjects (n = 12).

Keyword

Tadalafil; Pharmacokinetics; UPLC; Plasma

MeSH Terms

Ammonium Compounds
Humans*
Male
Methods*
Pharmacokinetics
Plasma*
Sildenafil Citrate
Tadalafil*
Sildenafil Citrate
Tadalafil

Figure

• 

  Figure 1 Collision-induced dissociation mass spectrum of (A) tadalafil and (B) sildenafil (internal standard, IS) in positive ion electrospray ionization mode. The MRM transitions of tadalafil and the IS that produced the m/z 268.3 and m/z 283.3 product-ion signals, respectively, were used for tadalafil quantification.

• 

  Figure 2 Representative MRM chromatograms (transition m/z 390.4 → 268.3 for tadalafil and m/z 475.3 → 283.3 for internal standard, IS) of blank plasma (A); blank plasma spiked with IS (B); LLOQ (5 ng/mL) plasma sample (C); a plasma sample 4 h after an oral administration of tadalafil 20 mg (D). Tadalafil (the blue line) and IS (the pink line).

• 

  Figure 3 Mean plasma concentrations (mean ± SD) vs. time profiles after following a single 20 mg tadalafil dose.

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