title: 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.affiliation: Department of Chemistry, Cleveland State University, 2399 Euclid Ave., Cleveland, OH 44115-2406, USA.
authors.forename: Yu-Chieh
authors.initials: YC
authors.lastname: Liao
authors.forename: Shuming
authors.initials: S
authors.lastname: Yang
authors.forename: Mei-Jywan
authors.initials: MJ
authors.lastname: Syu
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
mesh_terms: D019342:Acetic Acid / Q000032:analysis; D000970:Antineoplastic Agents / Q000032:analysis / Q000737:chemistry; D001547:Benzaldehydes / Q000032:analysis* / Q000737:chemistry; D002021:Buffers; D002138:Calibration; D002625:Chemistry, Organic; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry; D004260:DNA Repair; D004345:Drug Industry / Q000379:methods; D006898:Hydroxylamines / Q000032:analysis*; D008956:Models, Chemical; D055601:Organic Chemistry Phenomena; D015203:Reproducibility of Results; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D000970:Antineoplastic Agents; D001547:Benzaldehydes; D002021:Buffers; D006898:Hydroxylamines; D012997:Solvents; C055506:4-(diethylamino)benzaldehyde; D004247:DNA; C005214:methoxyamine; D019342:Acetic Acid