Validated LC-MS/MS method for the simultaneous determination of chlorambucil and its prodrug in mouse plasma and brain, and application to pharmacokinetics

J Pharm Biomed Anal. 2014 Oct:99:74-8. doi: 10.1016/j.jpba.2014.07.010. Epub 2014 Jul 17.

Abstract

A simple, sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous determination of chlorambucil (CHL) and the prodrug of chlorambucil (CHLS) in mouse plasma and brain tissue. Detection was performed on a Diamonsil ODS chromatography column using gradient elution with a mobile phase of 0.2% aqueous formic acid and acetonitrile. Mass spectrometry was carried out in multiple reaction monitoring mode using a positive electrospray ionization interface. Good linearity was found for CHLS and CHL in plasma and brain tissue in different linear ranges (r>0.9996). Intra-day and inter-day precision was within 9.11% and accuracy was not more than 11.07%. The validated method was successfully applied to the pharmacokinetic study of CHLS and CHL in mice after intravenous administration.

Keywords: Brain; Chlorambucil; LC–MS/MS; Pharmacokinetics; Prodrug.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents, Alkylating / administration & dosage
  • Antineoplastic Agents, Alkylating / blood*
  • Antineoplastic Agents, Alkylating / pharmacology
  • Brain / metabolism*
  • Chlorambucil / administration & dosage
  • Chlorambucil / blood*
  • Chlorambucil / pharmacokinetics
  • Chromatography, Liquid / methods
  • Dose-Response Relationship, Drug
  • Limit of Detection
  • Male
  • Mice, Inbred Strains
  • Molecular Structure
  • Prodrugs / administration & dosage
  • Prodrugs / pharmacokinetics*
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Tandem Mass Spectrometry / methods

Substances

  • Antineoplastic Agents, Alkylating
  • Prodrugs
  • Chlorambucil