A Novel Simultaneous Determination of Sarpogrelate and its Active Metabolite (M-1) in Human Plasma, Using Liquid Chromatography-Tandem Mass Spectrometry: Clinical Application.
10.3343/alm.2015.35.4.391
- Author:
Jeong Soo YANG
1
;
Jung Ryul KIM
;
Eungi CHO
;
Wooseong HUH
;
Jae Wook KO
;
Soo Youn LEE
Author Information
1. Clinical Trial Center, Clinical Research Institute, Samsung Medical Center, Korea.
- Publication Type:Original Article
- Keywords:
Clinical study;
Core-shell-type chromatography;
Metabolite;
Sarpogrelate;
Simultaneous determination
- MeSH:
Calibration;
Chromatography, Liquid;
Humans;
Mass Spectrometry*;
Particle Size;
Plasma*;
Tandem Mass Spectrometry
- From:Annals of Laboratory Medicine
2015;35(4):391-398
- CountryRepublic of Korea
- Language:English
-
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.
