Ultra-performance liquid chromatography/tandem mass spectrometric determination of diastereomers of SCH 503034 in monkey plasma

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-06, Vol.852 (1), p.92-100
• Main Authors: Wang, Ganfeng, Hsieh, Yunsheng, Cheng, Kuo-Chi, Morrison, Richard A., Venkatraman, Srikanth, Njoroge, F. George, Heimark, Larry, Korfmacher, Walter A.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chromatography, Liquid - methods; Diastereomers; Fundamental and applied biological sciences. Psychology; General pharmacology; LC/MS/MS; Macaca fascicularis; Male; Medical sciences; Pharmacology. Drug treatments; Plasma; Proline - analogs & derivatives; Proline - blood; Proline - chemistry; Reference Standards; Reproducibility of Results; SCH 503034; Sensitivity and Specificity; Stereoisomerism; Tandem Mass Spectrometry - methods; UPLC
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.12.038

This paper describes the development and qualification of a fast, sensitive and specific ultra-performance liquid chromatography/tandem mass spectrometric (UPLC/MS/MS) method for the determination of diastereomers of SCH 503034 in monkey plasma. The analytical method involves direct protein precipitation with a mixture of methanol/acetonitrile (10/90) containing an internal standard, followed by separation of the stereoisomers on an Acquity UPLC™ C 18 column and detected by selected reaction monitoring (SRM) in positive ionization mode using atmospheric pressure chemical ionization (APCI). The effects of ion-pairing agents on separation and ionization efficiency were investigated. The two diastereomers were well separated ( R = 1.3) with a runtime of 5 min under an isocratic condition. The method was qualified. The linear concentration range was 1–2500 ng/ml for the both stereoisomers. Inter-assay mean bias and relative standard deviation (R.S.D.) were in the range of −1.2% to 3.6% and 2.8–10%, respectively. Intra-assay mean bias and R.S.D. were in the range of −1.3% to 5.5% and 2.3–7.8%, respectively. Recoveries of the stereoisomers at concentration levels of 2.5, 50 and 1000 ng/ml were 87.2–90.0%, 89.1–90.4% and 92.3–94.3%, respectively. The LLOQ for this assay was 1 ng/ml. No matrix interferences were observed in six different sources of blank monkey plasma.