A comprehensive forced degradation studies of Cariprazine hydrochloride using LC-HRMS/MS and in silico toxicity predictions of its degradation products.

Cariprazine hydrochloride is a second-generation antipsychotic drug and has been approved by the US Food and Drug Administration and the European Medicines Agency. Herein, we have reported a stability-indicating assay method for the Cariprazine hydrochloride and characterization of its major degradation products using LC-HRMS/MS. The drug substance Cariprazine hydrochloride was subjected to acid, base, oxidation, thermal and photolytic stress degradation. A total of five novel degradation products (DP-1 to DP-5) of Cariprazine hydrochloride drug substance were formed under various acid, base and oxidative conditions. In silico toxicity of the degradation products was evaluated using Leadscope prediction platform. We successfully separated Cariprazine Hydrochloride peak from degradation products (DP-1 to DP-5) by using gradient elution on an Inertsil C18 column (150 × 4.6 mm, 5 µm). The drug substance was found to be labile to acidic, alkaline hydrolytic, and oxidative conditions and stable to photolytic and thermal stress conditions. The degradation pathways were delineated by explaining the putative mechanism of degradation in various conditions. The stability-indicating assay method was validated according to the ICH guideline and hence can be used for routine quality control and stability study analysis of Cariprazine hydrochloride in pharmaceutical industries and research laboratories.

Jain M ，Khan S 《-》

LC/Q-TOF-MS-based structural characterization of enasidenib degradation products and establishment of a stability-indicating assay method: Insights into chemical stability.

Enasidenib (EDB) is an orally active selective mutant isocitrate dehydrogenase-2 enzyme inhibitor approved by the U.S. Food and Drug Administration to treat acute myeloid leukemia. It lacks a reported forced degradation study and a stability-indicating assay method (SIAM). This study addresses this gap by establishing a degradation profile in accordance with the International Council for Harmonisation Q1A and Q1B (R2) guidelines and developing a validated SIAM for EDB. EDB was exposed to forced degradation under various conditions (hydrolytic, photolytic, oxidative, and thermal stress). Degradation samples were analyzed using high-performance liquid chromatography on an Agilent ZORBAX Eclipse Plus C18 column with a mobile phase consisting of 0.1% formic acid in Milli-Q water and acetonitrile at a flow rate of 1 mL/min and detection at 270 nm. Liquid chromatography-quadrupole time-of-flight-high-resolution mass spectrometry (LC/Q-TOF HRMS) was used for the identification and characterization of degradation products. Nitrosamine risk assessment was conducted using a modified nitrosation assay procedure (NAP) test due to the presence of a secondary amine group in the drug, which is liable to forming nitrosamine drug substance-related impurities (NDSRI). The drug exhibited significant degradation under acidic, basic, photolytic, and oxidative conditions in the solution state. A total of nine degradation products (DP) were formed (DP-I, DP-III, and DP-IV: acidic conditions; DP-I and DP-III: basic conditions; DP-II, DP-V, DP-VI, and DP-VII: oxidative stress; and DP-VII, DP-VIII, and DP-IX: photolytic conditions), which were separated and identified using reversed-phase high-performance liquid chromatography and characterized using liquid chromatography-tandem mass spectrometry. The mechanism behind the formation of EDB degradation products has been discussed, and this study was the first to develop a degradation pathway for EDB. In addition, the possibilities of NDSRI formation for EDB were studied using a modified NAP test, which can contribute to the risk assessment of the drug. Forced degradation studies were conducted by establishing a SIAM for EDB. All the degradation products were characterized by mass spectral data obtained using LC/Q-TOF-HRMS.

Chakkar A ，Chaturvedi S ，Rajput N ，Sengupta P ，Sharma N ... -  《-》

Identification and characterization of two new oxidation degradation impurities in cinnarizine through LC-HRMS/MS and (1)H NMR, along with in silico toxicity predictions of its degradation products.

Jain M ，Khan S 《-》

LC and LC-MS/MS studies for identification and characterization of new degradation products of ibrutinib and elucidation of their degradation pathway.

Forced degradation/stress degradation studies of ibrutinib drug were done in hydrolytic (acidic, alkaline and neutral), thermal, photolytic and oxidative degradation conditions in different temperature conditions as per International Conference on Harmonization (ICH) guideline Q1A(R2) in order to identify and characterize degradation products (DPs) of ibrutinib. The study revealed that ibrutinib is extremely sensitive to oxidative degradation even at room temperature. The drug substance is highly sensitive to alkaline hydrolysis and susceptible to acidic hydrolysis at 80 °C temperature condition, whereas found stable in neutral, photolytic and thermal stress conditions. Successful separation of ibrutinib and its ten degradation products formed during stress degradation condition were observed using Waters Acquity UPLC C-18 stationary phase (100 mm × 2.1 mm, 1.7 μm) with gradient elution using mobile phase consisting of Eluent-A: ammonium acetate (20 mm, pH-6) and Eluent-B: acetonitrile. The detection was carried out at 215 nm wavelength. Flow rate was set at 0.3 mL/min with injection volume of 5 μL. The drug substance degraded to one degradation product (DP-I) in acidic hydrolysis, five DPs (DP-I, DP-II, DP-V, DP-VIII and DP-IX) in basic hydrolysis and five DPs (DP-III, DP-IV, DP-VI, DP-VII and DP-X) in oxidative degradation condition. A novel and highly sensitive HRMS/MS/TOF method was developed to identify and characterize all the ten DPs formed during stress study. All the DPs were characterized using ESI positive mode. Except DP-I, all the degradation products formed were found to be new degradation impurities and their fragmentation pathways have never been reported earlier. The proposed mechanism and pathway of degradation products of ibrutinib were discussed and outlined.

Mehta L ，Naved T ，Grover P ，Bhardwaj M ，Mukherjee D ... -  《-》

Method validation and characterization of stress degradation products of azelastine hydrochloride using LC-UV/PDA and LC-Q/TOF-MS studies.

Azelastine HCl is a second-generation H1 -receptor antagonist approved by the US Food and Drug Administration (US FDA) for treating seasonal allergic rhinitis and non-allergic vasomotor rhinitis. This study encompasses the validation of a liquid chromatography-ultra violet photo diode array (LC-UV/PDA) method for the drug and its extension to liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q/TOF-MS) studies for identification and characterization of various stress degradation products of the drug. Stress degradation of azelastine HCl was undertaken under the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) prescribed conditions of hydrolytic, photolytic, oxidative, and thermal stress. The degraded drug solutions were analyzed using Ultra Performance Liquid Chromatography (UPLC) employing a C18 (100 × 4.6 mm; 2.6 μ, Kinetex) column by isocratic elution. Detection wavelength was 241 nm. The degradation products were identified and characterized using UPLC-MS/TOF studies, and an attempt was made to isolate one of the degradation products by solvent extraction. The drug was found to significantly degrade under acidic/alkaline/neutral photolytic, oxidative, and alkaline hydrolytic conditions. Six degradation products (I-VI) were identified through LC-Q/TOF-MS studies that were adequately resolved from the drug with the developed UPLC method. All degradation products (I-VI) were ionized in the total ion chromatogram (TIC) in the LC-MS studies, and these were identified and characterized, and the degradation pathway of the drug was postulated. One of the oxidation products isolated from the degraded drug solution was characterized through differential scanning calorimetry, Fourier-transform infrared spectroscopy, and nuclear magnetic resonance spectral data. Six degradation products generated from stress degradation studies on azelastine HCl were adequately resolved through LC-UV/PDA studies followed by method validation. These were successfully identified and characterized through LC-Q/TOF-MS studies, and the degradation pathways for the generation of these products from the drug have been postulated.

Shekhar S ，Bali A 《-》