Background:
A simple and sensitive Ultra Performance Liquid Chromatography-Mass Spectrometry
method was developed and validated to measure the concentrations of Alogliptin (ALO),
Linagliptin (LIN), Saxagliptin (SAX), and Sitagliptin (SIT) using Pioglitazone (PIO) as an internal
standard.
Methods:
Chromatographic separation of six gliptins was achieved on a C-18 column (100×2.1 mm,
2.7 μm) using a mobile phase consisting of formic acid in water, 0.1%v/v: acetonitrile in gradient elution.
Electrospray ionization (ESI) source was operated in the positive ion mode. Targeted MS/MS mode
on a QTOF MS was used to quantify the drug utilizing the transitions of 340.1(m/z), 473.2 (m/z), 316.2 (m/z),
408.1 (m/z), and 357.1 (m/z) for ALO, LIN, SAX, SIT and PIO respectively.
Results:
As per ICH Q2R1 guidelines, a detailed validation of the method was carried out and the
standard curves were found to be linear over the concentration ranges of 1516.0-4548.1 ng mL-1, 519.8-
1559.4 ng mL-1, 1531.4-4594.3 ng mL-1and 1519.6-4558.8 ng mL-1 for ALO, LIN, SAX and SIT respectively.
Precision and accuracy results were within the acceptable limits. The mean recovery was found to be
98.8 _ 0.76 % (GEM), 102.2 _ 1.59 % (LIN), 95.3 _ 2.74 % (SAX) and 99.2 _ 1.75 % (SIT) respectively.
Conclusions:
The optimized validated UPLC QTOF-MS/MS method offered the advantage of shorter
analytical times and higher sensitivity and selectivity. The optimized method is suitable for application
in quantitative analysis of pharmaceutical dosage forms for QC laboratory.
