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J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 1;1061-1062:123-127. doi: 10.1016/j.jchromb.2017.07.011. Epub 2017 Jul 8.

Development of a LC-MS/MS method for the determination of CKD-712 in rat plasma: Application to a pharmacokinetic study in rats.

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Department of Pharmacy, National University of Singapore, Singapore.
College of Pharmacy, Chungnam National University, Daejeon, Republic of Korea.
Korea Institute of Toxicology, Daejeon, Republic of Korea.
Graduate School of New Drug Discovery and Development, Chungnam National University, Daejeon, Republic of Korea.
Korea Institute of Toxicology, Daejeon, Republic of Korea. Electronic address:


CKD-712 is a potential treatment for sepsis, as it exhibits protective effects against lipopolysaccharide-mediated platelet aggregation, inducible nitric oxide synthase expression, and cecum-ligation puncture-induced septic mortality in mice. In this study, we develop a rapid and sensitive LC-MS/MS method for determining CKD-712 in rat plasma. CKD-712 and papaverine hydrochloride (an internal standard) were analyzed using an LC-MS/MS system consisting of an Agilent HPLC system (HP-1100) equipped with an Atlantis HILIC Silica (2.1×50mm, 3μm) column and a API 4000 (Applied Biosystems/MDS Sciex, USA) in a positive ESI mode. We utilized multiple reaction monitoring (MRM) at m/z transitions of 306.2-164.0 to analyze CKD-712, and 340.3-202.1 m/z for IS, with a mobile phase of acetonitrile (0.025% trifluoroacetic acid):20mM ammonium acetate (94:6, v/v) at a flow rate of 0.25mL/min. The lower limit of quantification (LLOQ) was 5ng/mL, with a linearity ranging from 5 to 1000ng/mL (r>0.999). Validation parameters including specificity, precision, accuracy, matrix effect, recovery, dilution effect and stability results were well within acceptance criteria, and applied successfully on a pharmacokinetic study in rats.


CKD-712; LC/MS/MS; Method validation; Pharmacokinetics; Rat plasma

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