Validation of a robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis
Susan C Scott1,2, Nicole M Anders1,2, Ping He2
1Department of Oncology, School of Medicine, Johns Hopkins University, Baltimore, MD, USA.
Abstract:
The Bcl-2 family small molecule inhibitor navitoclax is being clinically evaluated to treat multiple cancers including lymphoid malignancies and small cell lung cancer. A sensitive and reliable method was developed to quantitate navitoclax in human plasma using liquid chromatography with tandem mass spectrometry with which to perform detailed pharmacokinetic studies. Sample preparation involved protein precipitation using acetonitrile. Separation of navitoclax and the internal standard, navitoclax-d8, was achieved with a Waters Acquity UPLC BEH C18 column using isocratic flow over a 3 min total analytical run time. A SCIEX 4500 triple quadrupole mass spectrometer operated in positive electrospray ionization mode was used for the detection of navitoclax. The assay range was 5-5,000 ng/ml and proved to be accurate (89.5-104.9%) and precise (CV ≤ 11%). Long-term frozen plasma stability for navitoclax at -70°C was at least 34 months. The method was applied for the measurement of total plasma concentration of navitoclax in a patient receiving a 250 mg daily oral dose.
Insights
A new liquid chromatography-tandem mass spectrometry method accurately quantifies navitoclax in human plasma. This validated assay supports pharmacokinetic studies for navitoclax in cancer treatment.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Biochemistry
Background:
- Navitoclax, a Bcl-2 inhibitor, is under clinical evaluation for various cancers.
- Accurate quantification of navitoclax in biological matrices is crucial for pharmacokinetic studies.
Purpose of the Study:
- To develop and validate a sensitive and reliable bioanalytical method for quantifying navitoclax in human plasma.
- To support detailed pharmacokinetic studies of navitoclax in cancer patients.
Main Methods:
- Liquid chromatography with tandem mass spectrometry (LC-MS/MS) was employed.
- Sample preparation involved acetonitrile-mediated protein precipitation.
- Separation was performed on a Waters Acquity UPLC BEH C18 column with a 3-minute run time.
Main Results:
- The assay demonstrated a wide linear range (5-5,000 ng/ml).
- High accuracy (89.5-104.9%) and precision (CV ≤ 11%) were achieved.
- Navitoclax showed stability in plasma for at least 34 months at -70°C.
Conclusions:
- A validated LC-MS/MS method enables reliable quantification of navitoclax in human plasma.
- The method is suitable for pharmacokinetic profiling in clinical trials.
- This assay facilitates the clinical evaluation of navitoclax in cancer therapy.
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