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

Abstract

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.

Strategy to Assess and Control Nitrosamine Formation in API during Storage: A Sitagliptin Case Study

“…32−40 Recently, few specific methods (UHPLC-MS) for quantification of NTTP impurity in sitagliptin and in drug products were reported. 41,42 To the Sitagliptin, as an active pharmaceutical ingredient (API), contains an impurity, namely, triazolo pyrazine (Figure 2), that is susceptible to nitrosation, generating the nitrosamine impurity NTTP. Notably, this nitrosamine has been assigned a very low daily limit of 37 ng/day, in stark contrast to the majority of other nitrosamine drug substance-related impurities (NDSRIs) with a limit of 1500 ng/day.…”

Development and Validation of Analytical Method Using Gas Chromatography with Triple Quadrupole Mass Spectrometry for the Detection of Alkyl Halides as Potential Genotoxic Impurities in Posaconazole

“…Analytical scientists must develop the required techniques to precisely analyze and regulate the amounts of PGIs in drugs [22][23][24][25][26]. Appropriate analytical methods are necessary to develop reliable manufacturing processes and ensure patient safety.…”