Development and validation of an HPLC-UV method for the analysis of methoxyamine using 4-(diethylamino)benzaldehyde as a derivatizing agent

Yu Chieh Liao, Shuming Yang, Mei Jywan Syu, Yan Xu

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Methoxyamine (MX) is a potential new anti-cancer drug. In this paper, a quantitative HPLC-UV method for MX using 4-(diethylamino)benzaldehyde (DEAB) as a derivatizing agent has been developed and validated. The studies showed that MX reacts with DEAB under acidic conditions to form protonated 4-(diethylamino)benzaldehyde o-methyloxime (DBMOH+). The equilibrium between DBMOH+ and its conjugate base 4-(diethylamino)benzaldehyde o-methyloxime (DBMO) is affected by both buffer concentration and organic solvent content in the solution. The method developed uses a reversed phase C18 column for the separation of MX derivatives, an internal standard benzil for method calibration, and a UV detector at a wavelength of 310 nm for analyte detection. The MX derivatives can be resolved in ca. 20 min. The method has a linear calibration range from 0.100 to 10.0 μM with a correlation coefficient of 0.999 for MX and a detection limit of 5 pmol with a 50 μl sample size. The intra-assay and inter-assay precision expressed in terms of percent relative standard deviation were ≤5 and 8%; and the intra-assay and inter-assay accuracy defined as the measured value divided by the accepted value multiplied by 100% were 94.2-100 and 92.6-111%, respectively. This method may be used for the analysis of MX in pharmaceutical preparations.

Original languageEnglish
Pages (from-to)724-729
Number of pages6
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume39
Issue number3-4
DOIs
Publication statusPublished - 2005 Sep 15

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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