Ann Lab Med.  2015 Jul;35(4):391-398. 10.3343/alm.2015.35.4.391.

A Novel Simultaneous Determination of Sarpogrelate and its Active Metabolite (M-1) in Human Plasma, Using Liquid Chromatography-Tandem Mass Spectrometry: Clinical Application

• Yang JS
• Kim JR
• Cho E
• Huh W
• Ko JW
• Lee SY

• KMID: 2045860
• DOI: http://doi.org/10.3343/alm.2015.35.4.391

Abstract

BACKGROUND
This study describes a novel analytical method for simultaneously determining sarpogrelate and its metabolite (M-1) in human plasma, using liquid chromatography coupled with tandem mass spectrometry, with electrospray ionization in the positive ion mode.
METHODS
Sarpogrelate, M-1, and labeled internal standard (d3-sarpogrelate) were extracted from 50 microL of human plasma by simple protein precipitation. Chromatographic separation was performed by using a linear gradient elution of a mobile phase involving water-formic acid (99.9:0.1, v/v) and acetonitrile-formic acid (99.9:0.1, v/v) over 4 min of run time on a column, with a core-shell-type stationary phase (Kinetex C18, 50 mm x 2.1 mm i.d., 2.6-microm particle size, Phenomenex, USA). Detection of the column effluent was performed by using a triple-quadruple mass spectrometer in the multiple-reaction monitoring mode.
RESULTS
The developed method was validated in human plasma, with lower limits of quantification of 10 ng/mL for sarpogrelate and 2 ng/mL for M-1. The calibration curves of sarpogrelate and M-1 were linear over the concentration ranges of 10-2,000 and 2-400 ng/mL, respectively (R2>0.99). The carry-over effect, precision, accuracy, and stability of the method met the criteria for acceptance.
CONCLUSIONS
A simple, fast, robust, and reliable analytical method was successfully developed and applied to the high-throughput determination of sarpogrelate and its metabolite in real plasma samples in a pharmacokinetic study of healthy subjects.

Keyword

Clinical study; Core-shell-type chromatography; Metabolite; Sarpogrelate; Simultaneous determination

MeSH Terms

Calibration
Chromatography, Liquid
Humans
Mass Spectrometry*
Particle Size
Plasma*
Tandem Mass Spectrometry

Figure

• 

  Fig. 1 Representative multiple reaction monitoring chromatograms of sarpogrelate (left panel), its metabolite (M-1; middle panel), and internal standard (IS; right panel) in (A) double-blank plasma, (B) blank plasma spiked with IS, (C) blank plasma spiked with sarpogrelate and M-1 at lower limit of quantification (LLOQ; 10.0 and 2.0 ng/mL, respectively) and IS, and (D) real plasma sample spiked with IS.

• 

  Fig. 2 Mean plasma concentrations of (A) sarpogrelate and (B) its metabolite (M-1) after a single oral administration of 100 mg sarpogrelate in 19 healthy male subjects. Bars represent standard errors.

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