Abstract
A high-throughput method was developed and validated for the quantitative determination of MMI270B, an inhibitor of matrix metalloprotease (MMP) enzymes, in human plasma. The method was based on reverse-phase chromatographic separation of the analyte from plasma extract followed by atmospheric pressure chemical ionization (APCI) and tandem mass spectrometry in the selected reaction monitoring mode (SRM). Extraction was performed using simple protein fi ltration in the 96-well plate format to increase the throughput of the method. Optimised chromatographic separation in a short column (30 x 4.6 mm i.d.) coupled with positive APCI mode of ionization followed by selective SRM mode of detection yielded clean chromatograms with minimal signal suppression. The chromatographic conditions resolved isobaric interference peaks observed in samples from patients dosed with MMI270B. The standard curve was linear (r = 0.997) within the concentration range of 1.04 (lower limit of quanti fi cation) to 1040 ng/mL using 0.1 mL of human plasma. The accuracy of the method varied from 93.6 to 103% with a precision of 2.17-6.71% over the concentration range. The method was simple, rapid, and robust with an analyte recovery of >98%.
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