Abstract

DEVELOPMENT AND VALIDATION OF GC-MS METHOD FOR THE TRACE LEVEL DETERMINATION OF POTENTIAL GENOTOXIC IMPURITIES IN ANTI CANCER DRUG SUBSTANCE LENALIDOMIDE

Srivalli Kasina*, Annapurna Nowduri, Raghu Babu K. and Ramdas Chavakula

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Abstract

A Simple and trace level-sensitive Gas chromatography with mass spectrometer (GC-MS) method was developed, optimized and validated for the determination of seven potentially genotoxic impurities which are having structural alerts i.e., Methyl-2-methyl-3- nitrobenzoate (PGI-1), 4-Nitrophthalide (PGI-2), Methyl-2- Chloromethyl-3-nitrobenzoate (PGI-3), Methyl-2-bromomethyl-6- nitrobenzoate (PGI-4), Methyl-2-bromomethyl-3-nitrobenzoate (PGI- 5), Methyl-2-bromomethyl-4-nitrobenzoate (PGI-6), Methyl-2- bromomethyl-5-nitrobenzoate (PGI-7) in Lenalidomide (LND) drug substance. The chromatographic separation of seven genotoxic impurities were achieved on Capillary GC column, SPB-1, Fused silica capillary column; 30 m length; 0.25mm internal diameter, coated with 100% dimethyl polysiloxane stationary phase of 1.0 μm film thickness and passing helium carrier gas with Electron Impact ionization (EI) in Selective Ion Monitoring (SIM) mode by using liquid-liquid extraction sample preparation technique. The analytical method has been demonstrated through method validation experiments w.r.t. ICH guidelines, like specificity, linearity, Limit of Detection, Limit of Quantification, method precision and accuracy (recovery) parameters with allowable TTC limit. The mass fragments (m/z) were selected for the quantification of PGI-1 (m/z-178), PGI-2 (m/z-150), PGI-3 (m/z-161), PGI-4 (m/z-256), PGI-5 (m/z-162), PGI-6 (m/z-194) and PGI-7 (m/z-194). The established limit of detection values for the PGI-2 and PGI-4 is 0.05 μg mL−1 and for PGI-1, PGI-3, PGI-5, PGI-6 and PGI-7 is 0.037 μg mL−1. The established limit of quantitation values for the PGI-2 and PGI-4 is 0.15 μg mL−1 and for PGI-1, PGI-3, PGI-5, PGI-6 and PGI-7 is 0.11 μg mL−1. This developed method was found to be linear with correlation coefficient is greater than 0.990. The average recoveries for the accuracy were in the range of 84.2–107.6%. Developmental and Validation experiments were discussed in detail in this paper.

Keywords: Lenalidomide (LND), GC-MS, Potential Genotoxic impurities, Liquid-Liquid extraction and Method validation.