Ultra-performance liquid chromatography/tandem mass spectrometric determination of diastereomers of SCH 503034 in monkey plasma.
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.
Wang
Ganfeng
G
Department of Drug Metabolism and Pharmacokinetics, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USA. ganfeng.wang@spcorp.com
Hsieh
Yunsheng
Y
Cheng
Kuo-Chi
KC
Morrison
Richard A
RA
Venkatraman
Srikanth
S
Njoroge
F George
FG
Heimark
Larry
L
Korfmacher
Walter A
WA
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D008252:Macaca fascicularis; D008297:Male; D011392:Proline / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
C512204:N-(3-amino-1-(cyclobutylmethyl)-2,3-dioxopropyl)-3-(2-((((1,1-dimethylethyl)amino)carbonyl)amino)-3,3-dimethyl-1-oxobutyl)-6,6-dimethyl-3-azabicyclo(3.1.0)hexan-2-carboxamide; D011392:Proline