Transl Clin Pharmacol.  2019 Sep;27(3):98-106. 10.12793/tcp.2019.27.3.98.

Simultaneous quantification of ticagrelor and its active metabolite, AR-C124910XX, in human plasma by liquid chromatography-tandem mass spectrometry: Applications in steady-state pharmacokinetics in patients

• Chae SU
• Min KL
• Lee CB
• Huang Z
• Chang MJ
• Bae SK

• KMID: 2458949
• DOI: http://doi.org/10.12793/tcp.2019.27.3.98

Abstract

A sensitive and simple liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of ticagrelor and its active metabolite, AR-C124910XX from 50 µL human plasma using tolbutamide as an internal standard as per regulatory guidelines. Analytes in plasma were extracted by simple protein precipitation using acetonitrile, followed by chromatographic separation with an Acclaimâ„¢ RSLC 120 C₁₈ column (2.2 µm, 2.1 × 100 mm) and a gradient acetonitrile-water mobile phase containing 0.1% formic acid within 8 min. Mass spectrometric detection and quantitation were conducted by selected reaction-monitoring on a negative electrospray ionization mode with the following transitions: m/z 521.11 → 361.10, 477.03 → 361.10, and 269.00 → 169.60 for ticagrelor, AR-C124910XX, and tolbutamide, respectively. The lower limit of quantifications was 0.2 ng/mL with linear ranges of 0.2-2,500 ng/mL (r²â‰¥ 0.9949) for both analytes. All validation data, including selectivity, cross-talk, precision, accuracy, matrix effect, recovery, dilution integrity, stability, and incurred sample reanalysis, were well within acceptable limits. This assay method was validated using Kâ‚‚-EDTA as the specific anticoagulant. Also, the anticoagulant effect was tested by lithium heparin, sodium heparin, and K₃-EDTA. No relevant anticoagulant effect was observed. This validated method was effectively used in the determination of ticagrelor and its active metabolite, AR-C124910XX, in plasma samples from patients with myocardial infarction.

Keyword

AR-C124910XX; Human plasma; LC-MS/MS; Ticagrelor; Validation

MeSH Terms

Heparin
Humans*
Lithium
Mass Spectrometry*
Methods
Myocardial Infarction
Pharmacokinetics*
Plasma*
Tolbutamide
Heparin
Lithium
Tolbutamide

Figure

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  Figure 1 Product ion mass spectra of [M – H]− ions of ticagrelor (A), AR-C124910XX (B), and tolbutamide, IS, (C).

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  Figure 2 Representative chromatograms of ticagrelor (I), AR-C124910XX (II), and tolbutamide (III): (A) double blank plasma, (B) blank plasma spiked with LLOQ of ticagrelor and AR-C124910XX (0.2 ng/mL, both), (C) a plasma sample collected at 12 h after administration of 12 maintenance doses of ticagrelor on Day 7.

• 

  Figure 3 Mean plasma concentration-time profiles for ticagrelor (●) and AR-C124910XX (○) of four patients with myocardial infarction after oral administration of ticagrelor 180 mg loading dose (Day 1) followed by 90 mg twice/day maintenance dose on Day 7. Vertical bars represent standard deviation.

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