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

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 microM with a correlation coefficient of 0.999 for MX and a detection limit of 5 pmol with a 50 microl sample size. The intra-assay and inter-assay precision expressed in terms of percent relative standard deviation were < or =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.

• Authors: Yu-Chieh Liao, Shuming Yang, Mei-Jywan Syu, Yan Xu
• Journal: Journal of pharmaceutical and biomedical analysis (J Pharm Biomed Anal) Vol. 39 Issue 3-4 Pg. 724-9 (Sep 15 2005) ISSN: 0731-7085 [Print] England
• PMID: 15951147 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• Benzaldehydes
• Buffers
• Hydroxylamines
• Solvents
• 4-(diethylamino)benzaldehyde
• DNA
• methoxyamine
• Acetic Acid

Topics

• Acetic Acid (analysis)
• Antineoplastic Agents (analysis, chemistry)
• Benzaldehydes (analysis, chemistry)
• Buffers
• Calibration
• Chemistry, Organic
• Chemistry, Pharmaceutical (methods)
• Chromatography (methods)
• Chromatography, High Pressure Liquid (methods)
• Chromatography, Liquid
• DNA (chemistry)
• DNA Repair
• Drug Industry (methods)
• Hydroxylamines (analysis)
• Models, Chemical
• Organic Chemistry Phenomena
• Reproducibility of Results
• Solvents
• Spectrophotometry, Ultraviolet (methods)
• Temperature
• Time Factors
• Ultraviolet Rays