All regulatory organizations are paying close attention to the identification and measurement of genotoxic contaminants. Using conventional analytical techniques like high-performance liquid chromatography (HPLC) and gas chromatography to quantify probable genotoxic substances (PGIs) at the trace level is difficult (GC). Therefore, there is a necessity for advanced analytical techniques for the development of highly sensitive analytical procedures for the determination of trace-level PGIs in drug products and drug substances. This study’s goal is to develop and evaluate an analytical technique for measuring allyl chloride, a possible genotoxic contaminant in gemfibrozil. For the detection of very low and trace levels of impurities, a gas chromatography with a triple quadrupole mass spectrometry detector (GC-MS/MS) approach was developed and validated. Using a column USP phase G27, a nonpolar and low bleed 5% diphenyl, 95% dimethylpolysiloxane, with dimensions of 30 m in length, 0.32 mm internal diameters, and 1.5 m film thickness, along with a flow rate of 2.0 mL/min and Helium (He) as a carrier gas, this method uses a thermal gradient elution program. The method was calibrated with a linearity range from 30% to 150% concentration with respect to the specification level and achieved a limit of detection (LOD) and limit of quantification (LOQ) were 0.005 ppm and 0.01 ppm, respectively, for allyl chloride. According to current ICH requirements, the method was validated, and it was discovered to be specific, exact, accurate, linear, sensitive, tough, robust, and stable. This method is suitable for determining allyl chloride in the regular analysis of Gemfibrozil.

Effective risk management and control methods for potentially genotoxic impurities (PGIs), including alkyl halides, are of significant importance in the medicinal (pharmaceutical) sector. The three alkyl halides in posaconazole are PGIs. The detection and assessment of genotoxic substances is a top priority for all regulatory organizations. Quantifying PGIs at trace levels using standard analytical techniques, such as gas chromatography (GC) and high-performance liquid chromatography (HPLC), is challenging for the pharmaceutical manufacturing industry. Thus, the detection of trace quantities of PGIs in posaconazole is essential for developing sensitive analytical methodologies. The objective of this study was to establish an analytical technique for quantifying the three PGIs (alkyl halides) in posaconazole and its intermediate. These alkyl halides are 1-(2,4-difluorophenyl) ethan-1-one (PGI-1), (Z)-1-(1-bromoprop-1-en-2-yl)- 2,4-difluorobenzene (PGI-2), and 1-bromo-2-(2,4-difluorophenyl) propan-2-ol (PGI-3). To identify trace quantities (parts per million (ppm)) of these impurities, we employed a gas chromatography (GC-MS/MS) equipped with a triple quadrupole mass spectrometry detector. The GC column was a USP phase G43, which is a mid-polar 6% cyanopropyl; 94% polydimethylsiloxane, with a 60 m length, 0.32 mm inner diameter, and 1.8 μm film thickness. Helium (He) was used as the carrier gas, with a flow rate of 1.5 mL/min. A thermal gradient elution program was used for this procedure. The method was calibrated for the three PGIs with limits of detection (LOD) and quantification (LOQ) of 0.01 and 0.025 ppm, respectively. The linear range of concentrations (25–150%) was maintained with respect to the specification level. This method was validated according to the ICH regulations and was shown to be specific, rugged, robust, precise, sensitive, accurate, linear, and stable. Therefore, in this newly developed method, the combination of suitable analytical techniques, such as GC-MS/MS and proper chromatographic conditions and column selection with the lowest LOD and LOQ, have allowed the induction of excellent ionization. These conditions have successfully facilitated the identification of PGI-1, PGI-2, and PGI-3 in posaconazole and its intermediate during routine analysis.

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.

Cross contamination of β-lactams is one of the highest risks for patients using pharmaceutical products. Penicillin and some non-penicillin β-lactams may cause potentially life-threatening allergic reactions. The trace detection of β-lactam antibiotics in cleaning rinse solutions of common reactors and manufacturing aids in pharmaceutical facilities is very crucial. Therefore, the common facilities adopt sophisticated cleaning procedures and develop analytical methods to assess traces of these compounds in rinsed solutions. For this, a highly sensitive and reproducible ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the analysis of Cephapirin and Ceftiofur. As per the FDA guidelines described in FDA-2011-D-0104, the contamination of these β-lactam antibiotics must be regulated. The analysis was performed on an XBridge C18 column with 100 mm length, 4.6 mm diameter, and 3.5 µm particle size at an oven temperature of about 40 °C. The mobile phase was composed of 0.15% formic acid in water and acetonitrile as mobile phases A and B, and a flow rate was set to 0.6 mL/min. The method was validated for Cephapirin and Ceftiofur. The quantification precision and accuracy were determined to be the lowest limit of detection 0.15 parts per billion (ppb) and the lowest limit of quantification 0.4 ppb. This method was linear in the range of 0.4 to 1.5 ppb with the determination of coefficient (R2 > 0.99). This sensitive and fast method was fit-for-purpose for detecting and quantifying trace amounts of β-lactam contamination, monitoring cross contamination in facility surface cleaning, and determining the acceptable level of limits for regulatory purposes.