Go to contents (site navigation)

 


Realised by ALMS™
developer of the AIDS-HIV Reference project
Abstract No.: MoP-105
Session: High Throughput Analysis: From Discovery to Clinical Applications
Presentation date: Mon, Aug 28, 2006
Presentation time: 09:50 – 11:20

Quantitative Determination of CKD501 by High Turbulence Liquid Chromatography - Tandem Mass Spectrometry in Human Plasma and Urine

Hyun Suk Shin1, Hyo Bum Seo1, Jae Soo Shin2, Hong Woo Lee2, Soon Kil Ahn2, Joo-Youn Cho1, In Jin Jang1

1 Seoul National University College of Medicine, Seoul, Korea
2 Chong Kun Dang Research Institute, Chonan, Korea

Correspondence address: Hyun Suk Shin, Seoul National University College of Medicine, Pharmacology, 28 Yongon-Dong, Chongno-Gu, Seoul, 110-744 Korea.

Keywords: Collision-Induced Dissociation (CID); Electrospray Ionization (ESI); High Throughput; MS/MS, Liquid Chromatography.

Novel aspect: High-throughput liquid chromatography/tandem mass spectrometry (LC/MS/MS) method, which combines on-line sample extraction through high turbulence chromatography (HTLC), has been developed for CKD501 in human plasma and urine.

 

CKD501 was developed to treat type 2 (noninsulin-dependent) diabetes. To evaluate possible pharmacokinetic determinations of its response variability, a high-throughput liquid chromatography/tandem mass spectrometry (LC/MS/MS) method, which combines on-line sample extraction through high turbulence chromatography (HTLC), has been developed for CKD501 in human plasma and urine. The HTLC technology significantly reduced the time required for sample extraction and overall times for analysis of plasma and urine samples were 7 and 7.5 min, respectively. The analysis was performed on Luna C18, 3 µm (50 mm * 2.0 mm i.d.) with an acetonitrile-water mixture (with containing 0.1% formic acid) as a mobile phase. The flow rate was set at 0.25 mL/min and glipizide was used as the internal standard. The linear range of calibration curve for human plasma and urine are 0.5 1000 ng/mL, 0.2 - 250 ng/mL, respectively (coefficients of correlation >0.9996). The intra- and inter-day accuracy and precision were evaluated by analyzing five replicates of quality control samples. Accuracy and precision variations for plasma and urine are lower than 12 %. The method validated for extraction stability, room temperature stability, storage stability, Freeze/Thaw stability, assay specificity, and recovery. The stability of the technique was demonstrated by applying this method to analyze over 2000 plasma samples and 450 urine samples.