Development of a liquid chromatography–electrospray mass spectrometric method for the simultaneous analysis of benzoxazolinones and their degradation products

A new method for the simultaneous analysis of some benzoxazolinones, aminophenoxazinones and malonamic acids was developed based on liquid chromatography (LC) coupled to mass spectrometry (MS), using electrospray ionization (ESI) and operating in positive mode. Different ESI-MS parameters, such as fragmentor voltage, capillary voltage, drying gas flow, nebulizer gas pressure and drying gas temperature, were optimized in order to obtain structural information and to achieve maximum sensitivity. Chromatographic separation was performed by a reversed-phase LC column using a linear gradient of water and methanol. Quality assurance of the developed method was assessed by measuring parameters as linearity, sensitivity, repeatability and reproducibility. Quantification method based on the use of internal standard was developed, selecting synthetic 2-methoxy-2H-1,4-benzoxazin-3(4H)-one as internal standard. Good correlations were obtained for all analytes relative to this compound in the range of 0.05–1.5 ng/μL. Instrumental detection limits were between 0.02 and 0.2 ng/μL. Repeatability and reproducibility studies showed acceptable coefficient of variation values.