UPLC–MS/MS assay of 21-aminosteroid (lazaroid U74389G) for application in pharmacokinetic study

摘要: Lazaroids are potent inhibitors of lipid peroxidation, both in vitro and vivo. Additionally, a member the lazaroid family, U-74389G (LAZ) has been shown to have specific radio-protective anti-proliferative effects. However, there is no quantitative analytical method developed for measuring therapeutic levels LAZ aforementioned This article highlights development validation sensitive UPLC-MS/MS quantification LAZ, its subsequent application pharmacokinetic studies rats with lower limit (LLOQ) 1.95 ng/mL. internal standard diadzein (IS) were separated using ACQUITY UPLC(®) BEH C18 column. Gradient elution was used at flow rate 0.45 mL/min mobile phases consisting 0.1% formic acid water acetonitrile. (m/z 612→260) IS 255→199) detected by electrospray ionization (ESI) multiple reaction monitoring (MRM) positive mode on QTRAP(®) 5500 System. The validated as per US FDA Guidelines Bio-analytical Validation. extracted from rat plasma (100 μL) protein precipitation acetonitrile mean recovery matrix factor range 47.7-56.1%, 85.6-89.4%, respectively. calibration curve linear 1.95-250 inter-day intra-day accuracy precision values LLOQ, low, medium, high very concentration QC samples within ±15%. tested under different storage conditions, short-term bench-top stability (1h 3h 25°C), long-term (1 month -80°C), freeze-thaw cycle 3 cycles) processed auto-sampler (24h 10°C) ±15% nominal concentrations. further applied study after single intravenous dose 5 mg/kg.