Development and validation of an ultra-performance liquid chromatography method coupled with tandem mass spectrometry for determination of alizapride in human plasmaElectronic supplementary information (ESI) available. See DOI: 10.1039/c5ra10386j

The present study describes a novel liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the estimation of alizapride in human plasma by electro spray ionization in the positive mode using triple quadrupole mass spectrometry using midodrine as an internal standard (IS). Sample pretreatment was carried out with solid-phase extraction using Bond Elut C18 cartridges, resulting in an average recovery of 86.45 ± 0.62 of the investigated compound. The chromatographic separation was performed on an Acquity UPLC BEH shield reversed phase C18 column, using a gradient mobile phase consisting of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.2 mL min −1 . The molecular ion of the analyte was detected in positive ionization by multiple reaction monitoring (MRM). The mass transitions of m / z 316.24 → 124.19 and m / z 255.16 → 180.19 were used for detection of alizapride and its internal standard, respectively. The assay exhibited linear ranges from 1 to 1000 ng mL −1 for the analyte in human plasma. The LC-MS/MS method was fully validated for all the other parameters such as selectivity, linearity and range, LLOQ, precision and accuracy, matrix effect, recovery and stability. The lower limit of quantification (LLOQ) of the developed assay method for alizapride was 1 ng mL −1 . The intraday and interday variation of the current assay was evaluated with CV% within 4.8%, whereas the mean accuracy ranged from 93.59-100.19%. The samples were stable under the storage conditions for at least a month. In conclusion, the findings of the present study revealed the selectivity and sensitivity of this method for the estimation of alizapride in human samples. The proposed method was successfully applied to determine alizapride in human plasma samples after intramuscular administration of the drug. Representative (a) blank plasma samples, (b) spiked plasma sample at LLQQ (1 ng mL −1 ) with the IS, (c) spiked plasma sample at QCH (800 ng mL −1 ) with the IS and (d) real plasma sample at 10 minutes spiked with the IS.