Simultaneous Estimation of Genotoxic Impurities 4-Nitrobenzene Trifluoride, 4-Aminobenzene Trifluoride, and Benzo Trichloride in Cinacalcet Hydrochloride by GC-MS/MS Method

The main objective of this study is to develop and validate a novel, fast, and more sensitive gas chromatography-mass spectrometry (GC-MS/MS) method for the simultaneous estimation of 4-Nitrobenzotrifluoride (4-NBTF), 4-Amino-benzotrifluoride (4-ABTF), and Benzotrichloride (BTC) impurities in Cinacalcet hydrochloride (CH). The chromate-graphic separations were performed on a DB-624, 30m × 0.32mm × 1.8µm column with injector temperature of 150°C, and mode of injection is split with asplit ratio 1: 20. The carrier gas used was helium with a flowrate 1.5 mL/min, and theinjection load was 1.0 µL. Mass spectrometry quantitation was achieved by aquadrupole analyser with EI (Electron Ionization) ion source atasource temperature of 250ºC and interface temperature of 250ºC. The retention times for 4-NBTF, 4-ABTF, and BTC were at 6.13, 6.74and 7.64min, respectively. The calibration curve was linear over the concentration ranging from LOQ level to 150 % level with the correlation coefficient (r) of > 0.99. The percentage recovery was found to be within the specified range, i.e., 70.0 to 130.0 for the three impurities. The limit of detection (LOD) was established to 0.19, 0.19, and 0.18 ppm, whereas thelimit of quantification (LOQ) was obtained to 1.14, 1.12, and 1.11 ppm for 4-NBTF, 4-ABTF, and BTC, respectively. The test solutions with impurities were found to be stable in the diluent for 24 hours. A simple GC-MS/MS method was developed and validated for simultaneous estimation of three impurities in CH. The method was accurate, precise, linear, specific, sensitive, robust, and rugged as per ICH guidelines. The method has been applied to the real-time batch analysis and found to be suitable for routine quality control analysis of CH.

Development and Validation of Three Potential Genotoxic Impurities by Liquid Chromatography Single Quad Mass Detector in Iomeprol

A liquid chromatography with single quadrupole mass detection method was developed for the determination of potential genotoxic impurities (PGIs) in the Iomeprol active pharmaceutical ingredient. Chromatographic separation was achieved on an Agilent Eclipse plus C8 column (100 mm x 2.1 mm x 1.8 μm) with 0.1% formic acid in water as mobile phase A and acetonitrile as mobile phase B in gradient elution mode at a 0.1 mL/min. Executed validation summary demonstrated that the mass detection method had highly sensitive and selective. A linear calibration curve (correlation coefficient, r> 0.999) was attained at the concentration range of 0.1-125 ppm for three PGI’s. The Limit of Detection of Imp-A, Imp-B and Imp-C in drug substance of Iomeprol is 0.05 ppm. The accuracy was confirmed by calculated recoveries (98.4-101.5%). The precision was tested at three levels: injection repeatability, analysis repeatability and intermediate precision. The calculated relative standard deviations were within the specification. The developed method was able to quantitate all three PGI’s at a concentration level of 1 µg/mL.

A Simple and Sensitive LC-MS/MS Method for Determination and Quantification of Potential Genotoxic Impurities in the Vismodegib Active Pharmaceutical Ingredient

A rapid and sensitive LC-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitative analysis of four potential genotoxic impurities Imp-A (2-chloro-5-nitroaniline), Imp-B (1-chloro-2-iodo-4-nitrobenzene), Imp-C (1-(2-chloro-5-nitrophenyl)ethan-1-one) and Imp-D (2-chloro-5-nitrobenzoic acid) in Vismodegib API drug sample. This trace analysis was achieved on CSH C18, 15.0 cm x 3.0 mm, 1.7 micron column maintained at 45°C. Optimal mobile phase consisted of 0.05% formic acid in water was used as mobile phase A and acetonitrile used as mobile phase B in gradient mode with the flow rate of 0.5 mL/min. The developed method had the short run time of 12 minutes. Quantification of four potential genotoxic impurities were carried out using mass detection with electrospray ionization in multiple reaction monitoring mode. The method was linear in the range of 0.03 ppm to 1.50 ppm for four potential genotoxic impurities with a correlation coefficient not less than 0.999. The recoveries were found satisfactory over the range between 96.67 and 106.90% for all selected impurities. The developed method was able to quantitate all four PGIs at a concentration level of 0.03 ppm (0.03 ppm with respect to 20 mg /mL Vismodegib).

Method Development for the Control of Potential Genotoxic Impurities in Vigabatrin Using Gas Chromatography Techniques and Mass Spectroscopy Detector

Aim: To develop a sensitive headspace GC-MS method for the determination of potential genotoxic impurities in Vigabatrin. Place and Duration of Study: The study was performed in SIONC Pharmaceuticals, Visakhapatnam from June 2020 to March 2021. Methodology: The impurities were determined by selected ion monitoring mode using VF -WAXms (30 mts length, 0.25 mm internal diameter, 1.0 µ film thickness) column. Helium gas was used as carrier gas with a column flow of 1.0 mL/min. and injector temperature maintained at 220 0C. Oven Temperature, loop temperature and transfer line temperature were maintained in the head space at 70oC, 90oC and 100oC respectively. Results: The linearity of the method was proposed in the range of LOQ to 150 % for the genotoxic impurities by subjecting the data obtained to statistical analysis using linear regression model (r2 > 0.99). The method also gave acceptable recovery of all the four impurities at each level and was found to be accurate. The % RSD obtained in the method precision and intermediate precision were less than 11% depicting the precision of the method. The LOD and LOQ values were calculated based on the signal to noise ratio and are indicating the sensitivity of the method. The specificity of the method was checked for blank interference at the retention time of respective impurities. Conclusion: The results proved that the proposed headspace GC-MS method for the study of potential genotoxic impurities of Vigabatrin was sensitive, precise and accurate and could be routinely used in the quality control testing of the active pharmaceutical ingredient.

A Brief Review on Genotoxic impurities in Pharmaceuticals

Genotoxic impurities (GTIs) in pharmaceuticals are increasing concern to pharmaceutical industries due to their potential for human carcinogenicity. The literature currently lacks the Guidance for the analytical determination of diverse classes of GTIs. This review article provides brief information regarding Genotoxic impurity (GTI), its sources, their classification, and existing regulatory approaches to control Genotoxic impurities in Pharmaceuticals, also information regarding different types of GTI and examples of each class. It is very difficult for researchers to detect GTI at Trace level so the detection method is also given in the chart, almost all the GTI are Mutagenic but as shown in Nitrosamines it is Mutagenic and its carcinogenicity is also proved but there are no such strong evidence and literature which shows genotoxicity. So, it is included as Mutagenic & carcinogenic. Different Control strategies to control Genotoxicity are also discussed.