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A rapid ultra HPLC-MS/MS method for the quantitation and pharmacokinetic analysis of 3-deazaneplanocin A in mice
Cody J Peer1, Mahadev Rao, Shawn D Spencer
1Clinical Pharmacology Program, Office of the Clinical Director, National Cancer Institute, Bethesda, MD, USA.
Abstract:
3-Deazaneplanocin A (DZNep) has been shown to have anti-cancer activity in numerous cancer types and its continued preclinical, and eventual clinical, drug development will require rapid and sensitive bioanalytical methods in order to quantitate this drug for pharmacokinetic analyses. The ultra HPLC with positive thermospray tandem mass spectrometric (LC-MS/MS) detection affords the most sensitive (limit of quantitation 5ng/mL) and rapid (3min run time) bioanalytical method to date for DZNep. Due to the polar nature of this drug and the internal standard (tubercidin), a hydrophilic-interaction column (HILIC) was used. The method was accurate, with less than 10% deviation from nominal values, as well as precise, where both within-day and between-day precisions were less than 15%. A liquid-liquid extraction procedure was able to recover ∼90% of drug from a small volume (50microL) of mouse plasma. This method was successfully applied to a pharmacokinetic study in mice intravenously injected with DZNep.
Insights
A new ultra HPLC-MS/MS method quantitates 3-Deazaneplanocin A (DZNep) in mouse plasma. This rapid and sensitive bioanalytical technique supports preclinical and clinical drug development for DZNep's anti-cancer applications.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Biochemistry
Background:
- 3-Deazaneplanocin A (DZNep) exhibits anti-cancer properties across various cancer types.
- Effective preclinical and clinical development necessitates sensitive bioanalytical methods for pharmacokinetic analysis of DZNep.
Purpose of the Study:
- To develop and validate a rapid and sensitive bioanalytical method for quantifying DZNep in biological matrices.
- To support pharmacokinetic studies of DZNep in preclinical models.
Main Methods:
- Ultra-high-performance liquid chromatography coupled with positive thermospray tandem mass spectrometry (LC-MS/MS).
- Utilized a hydrophilic-interaction column (HILIC) due to the polar nature of DZNep and the internal standard (tubercidin).
- Developed a liquid-liquid extraction for sample preparation from mouse plasma.
Main Results:
- Achieved a limit of quantitation of 5 ng/mL for DZNep.
- Method demonstrated high sensitivity and a short run time of 3 minutes.
- Accurate ( <10% deviation) and precise ( <15% within-day and between-day) quantification.
- Liquid-liquid extraction yielded approximately 90% recovery from 50 µL of mouse plasma.
- Successfully applied the method to a mouse pharmacokinetic study.
Conclusions:
- The developed LC-MS/MS method is sensitive, rapid, accurate, and precise for DZNep quantification.
- This validated bioanalytical method is suitable for supporting pharmacokinetic studies in drug development.
- The method facilitates the advancement of DZNep as a potential anti-cancer therapeutic.
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