Liquid chromatography–electro spray ionization–tandem mass spectroscopy method for the quantification of alogliptin in spiked human plasma

Background
Alogliptin is the inhibitor of dipeptidyl peptidase-4. It acts to regulate blood sugar by increasing the amount of insulin in the body.
Aim
A liquid chromatography–mass spectroscopy/mass spectroscopy method was developed for the estimation of alogliptin in spiked human plasma. Liquid–liquid extraction technique was adopted for the extraction of alogliptin from human plasma.
Materials and methods
The chromatographic separation was performed on a Waters Symmetry shield RP C-18 column with 4.6-mm internal diameter, 5-μm particle size, 100A° pore size analytical column at a flow rate of 1.0 ml/min using a mixture of 0.3% formic acid and acetonitrile in the ratio of 20 : 80% v/v as mobile phase. Positive ion mode was selected to obtain the product ion m/z +339.19 (parent) →245.11 (product) for alogliptin and m/z +303.39 (parent) →232.16 (product) for internal standard.
Results
The developed method was satisfactorily validated as per United State Food and Drug Administration guidelines for the bioanalytical study because it exhibits excellent intraday and interday accuracy with % nominal 91.06→98.48% and precision percentage coefficient variation less than or equal to 2% in all quality control levels. Alogliptin revealed its linearity with correlation coefficient (=0.99) in the concentration range of 40.17–16 096 ng/ml, showed acceptable % extraction recovery (96.18→98.36%), and showed excellent matrix and analyte selectivity (% interference=0), as well as matrix effect (matrix factor 0.931 at lower quantitation limit and 1.14 at high quality control level). The stability study results of bench top, freeze thaw, autosampler, and short-term and long-term showed the accuracy in the range of 92.52–99.16% and percentage coefficient variation in the range of 0.22–1.93.
Conclusion
The method was successfully optimized and validated as per the United State Food and Drug Administration bioanalytical method development guidelines. The applicability of the developed method undoubtedly can further extend during preclinical and clinical trials.