Development and Validation for Quantification of 7-Nitroso Impurity in Sitagliptin by Ultraperformance Liquid Chromatography with Triple Quadrupole Mass Spectrometry

• Published in: Molecules (Basel, Switzerland) Switzerland), 2022-12, Vol.27 (23), p.8581
• Main Authors: Chittireddy, Hari Naga Prasada Reddy, Kumar, J V Shanmukha, Bhimireddy, Anuradha, Shaik, Mohammed Rafi, Khan, Merajuddin, Adil, Syed Farooq, Khan, Mujeeb, Aldhuwayhi, Fatimah N
• Format: Article
• Language: English
• Subjects: 7-nitroso impurity; Chromatography; Chromatography, High Pressure Liquid - methods; Chromatography, Liquid; Correlation coefficient; Correlation coefficients; Diabetes mellitus (non-insulin dependent); Diabetes Mellitus, Type 2; Flow velocity; Formic acid; Genotoxicity; Humans; ICH; Impurities; Linearity; Liquid chromatography; Mass spectrometry; Mass spectroscopy; nitrosamine; Optimization; Particle size; Pyrazine; Quadrupoles; Regulatory agencies; Reproducibility of Results; Retention; Ruggedness; Sitagliptin; Sitagliptin Phosphate; Solvents; Spectroscopy; Systems stability; Tandem Mass Spectrometry - methods; Type 2 diabetes; UHPLC-MS/MS
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27238581

The purpose of this research study was to develop an analytical method for the quantification of 7-nitroso-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4] triazolo [4,3-a] pyrazine (7-nitroso impurity), which is a potential genotoxic impurity. Since sitagliptin is an anti-diabetic medication used to treat type 2 diabetes and the duration of the treatment is long-term, the content of nitroso impurity must be controlled by using suitable techniques. To quantify this impurity, a highly sensitive and reproducible ultraperformance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed. The analysis was performed on a Kromasil-100, with a C18 column (100 mm × 4.6 mm with a particle size of 3.5 µm) at an oven temperature of approximately 40 °C. The mobile phase was composed of 0.12% formic acid in water, with methanol as mobile phases A and B, and the flow rate was set to 0.6 mL/min. The method was validated according to the current International Council for Harmonisation (ICH) guidelines with respect to acceptable limits, specificity, reproducibility, accuracy, linearity, precision, ruggedness and robustness. This method is useful for the detection of the impurity at the lowest limit of detection (LOD), which was 0.002 ppm, and the lowest limit of quantification (LOQ), which was 0.005 ppm. This method was linear in the range of 0.005 to 0.06 ppm and the square of the correlation coefficient (R2) was determined to be > 0.99. This method could help to determine the impurity in the regular analysis of sitagliptin drug substances and drug products.