Ultra-Sensitive LC-MS/MS Method for the Trace Level Quantification of Six Potential Genotoxic Nitrosamine Impurities in Telmisartan

Abstract

Nitrosamine impurities are potentially genotoxic which are considered under cohort of concern as per ICH M7 guidelines and need to be controlled at trace levels during quantification in drug substances and drug products for safe human consumption. Recent regulatory requirements also suggest the need to have highly sensitive analytical methods for the accurate quantification of Nitrosamine impurities. In this paper we have presented simple, rapid and ultra-sensitive LC-MS/MS method for six potential genotoxic nitrosamine impurities: N-Nitroso dimethyl amine (NDMA), N-Nitroso diethyl amine (NDEA), N-Nitroso Ethyl Iso propylamine (NEIPA), N-Nitroso-Nmethyl-4-aminobutyric acid (NMBA) N-Nitroso diisopropylamino (NDIPA) and N-Nitroso dibutyl amine (NDBA) with a LOQ of 0.004 ppm. Chromatographic separation is achieved using Zorbax SB C18 150 × 3.0 mm, 3.5 µ column with 0.1% formic acid in water as mobile phase A and 0.1% formic acid in methanol as mobile phase B at a flow rate of 0.3 ml/min using gradient mode of elution at a total run time of 18 minutes. Six nitrosamine impurities are successfully ionized and quantified in positive mode of atmospheric pressure chemical ionization (APCI) using multiple reaction monitoring (MRM). Method validation is performed as per ICH guidelines evaluating the limit of quantification and detection and found to give good S/N ratios with good linearity range of 0.002 -2 ppm with regression coefficient >0.99 for all the six nitrosamine impurities. Method recoveries are established using three-step sample preparation protocol and are found to be satisfactory within 80% -120%. The method can be used routinely applied for the detection of Nitrosamines in Telmisartan at a concentration of 1.5 ng/ml (0.03 ppm with respect to telmisartan concentration of 50 mg/ml).

Determination of Nitrosamines in Various Pharmaceuticals at Variable Temperatures

“…Guidance on controlling those impurities in medicinal and pharmaceutical products has been established by regulatory authorities like the USFDA and EMA due to the presence of different NA impurities (Chidella et al, 2021). These guidelines provide insights into the conditions that can contribute to the occurrence of NA impurities in pharmaceutical products.…”

“…Several NAs, including NDMA, N-nitroso-N-methyl-4aminobutyric acid (NMBA), N-nitroso diethylamine (NDEA), N-nitroso diisopropylamine (NDIPA), N-nitroso ethyl isopropylamine (NEIPA), N-nitroso dibutylamine (NDBA), N-nitroso ethyl methylamine (NMEA), N-nitroso pyrrolidine (NPyR), and N-nitroso piperidine (NPIP) (Table 1), may be encountered as impurities in drug groups with this molecular structure (Li et al, 2021;Hu et al, 2021). Assessment reports released by the USFDA and European Medicines Agency (EMA) enounce that NDMA, NDEA, NMBA, NDBA, N-methyl-N-nitrosoaniline (NMPhA) are the most commonly detected impurities in specific drugs (Chidella et al, 2021;USFDA, 2021;EMA, 2021). Consequently, detecting NAs has become a crucial issue in drugs prone to impurities due to their susceptible production processes and materials.…”

Nitrosamine Impurities in Pharmaceutical Dosage Form: A Review

“…The Telmisartan [11] LC-MS/Mass Spectroscopy Column: Zorbax SB C18 150×3.0mm, 3.5μm Mobile phase A was composed of 0.1% formic acid in water Mobile phase B was made up of 0.1% formic acid in methanol Flow rate: 0.3 ml/min [11] 2.…”

Determination of Genotoxic Impurity N-Nitroso-N-methyl-4-aminobutyric Acid in Four Sartan Substances through Using Liquid Chromatography–Tandem Mass Spectrometry

“…Additionally, an LC-MS/MS method was established for NMBA detection in losartan potassium, in which the detection time of NMBA was 2.868 min, the LOQ was 1.0 ng/mL and the RSD of peak areas for six groups of parallel samples was 4.77% [ 10 ]. Furthermore, Chidella et al developed an LC-MS/MS quantitative analysis method for six potentially genotoxic N-nitrosamine impurities in telmisartan, in which the running time was approximately 13.5 min and a relative LOQ was 0.004 ppm [ 11 ]. However, there have been no reports of the trace analysis of NMBA in candesartan cilexetil, olmesartan medoxomil, irbesartan and valsartan through using the LC-MS/MS method to date.…”