Transl Clin Pharmacol.  2017 Dec;25(4):183-189. 10.12793/tcp.2017.25.4.183.

Development and validation of analytical method for the determination of radotinib in human plasma using liquid chromatography-tandem mass spectrometry

• Seo HB
• Cho S
• Yoon YR
• Yim DS

• KMID: 2407000
• DOI: http://doi.org/10.12793/tcp.2017.25.4.183

Abstract

This study describes the development of an analytical method to determine radotinib levels in human plasma using high performance liquid chromatography (HPLC) coupled with triple quadrupole tandem mass spectrometry (MS/MS) for pharmacokinetic application. Plasma samples were sequentially processed by liquid-liquid extraction using methyl tert-butyl ether, evaporation, and reconstitution. Analytes were separated and analyzed using HPLC-MS/MS in selected reaction monitoring mode, monitoring the specific transitions of m/z 531 to 290 for radotinib and m/z 409 to 238 for amlodipine (internal standard). The HPLC-MS/MS analytical method was validated with respect to selectivity, linearity, sensitivity, accuracy, precision, recovery, and stability. Calibration curves were linear over a concentration range 5-3,000 ng/mL with correlation coefficients (r) > 0.998. The lower limit of quantification for radotinib in plasma was 5 ng/mL. The accuracy and precision of the analytical method were acceptable within 15% at all quality control levels. This method was suitable to determine radotinib levels in human plasma because of its simplicity, selectivity, precision, and accuracy.

Keyword

Radotinib (IY5511); HPLC-MS/MS; Human plasma; Method validation; Pharmacokinetic study

MeSH Terms

Amlodipine
Calibration
Chromatography, Liquid
Ether
Humans*
Liquid-Liquid Extraction
Mass Spectrometry*
Methods*
Plasma*
Quality Control
Tandem Mass Spectrometry
Amlodipine
Ether

Figure

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  Figure 1 Chemical structures of (a) radotinib (IY5511) and (b) amlodipine (IS) and collision-induced dissociation spectra of the corresponding protonated molecular ions, [M+H]+.

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  Figure 2 Determination of void volume using imatinib as a marker.

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  Figure 3 Typical SRM chromatograms of (a) a human blank plasma, (b) a human blank plasma spiked with IS (6 µg/mL), (c) a plasma spiked with radotinib (5 ng/mL) and IS (6 µg/mL), and (d) a plasma sample taken from a patient three hours after oral administration of a 400-mg radotinib and spiked with IS. The following transitions were monitored: m/z 531 → 290 for radotinib (left) and m/z 409 → 238 for IS (right).

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  Figure 4 Carryover effect: SRM chromatograms of (a) a radotinib calibration standard at the ULOQ (3 µg/mL) in human plasma and (b) a blank plasma sample right after the analysis of a ULOQ sample. The following transitions were monitored: m/z 531 → 290 for radotinib (left) and m/z 409 → 238 for IS (right).

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  Figure 5 Representative plasma concentration-time prole of radotinib after administration of 400-mg radotinib two times a day.

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