Pavithra Modachakanahally K, G Chaya, Deepakumari Hemavathi N, Revanasiddappa Hosakere D, Mohammed Salah Jasim, Majdi Hasan Sh, Alsabhan Abdullah H, Ukkund Shareefraza J
Department of Chemistry, Bharathi College Bharathinagara Mandya 571 422 Karnataka India
Department of Chemistry, Regional Institute of Education (NCERT) Bhubaneswar 751022 Odisha India
RSC Adv. 2024 Nov 1;14(47):34868-34882. doi: 10.1039/d4ra06299j. eCollection 2024 Oct 29.
This study examined the fragmentation, degradation pathways and DPs of pirtobrutinib, which have not been previously reported in the literature. The main goal of the current work is to develop, validate, and characterize forced degradation products using LC-MS/MS. An isocratic HPLC methodology was developed for the quantitative measurement of pirtobrutinib at a of 219 nm. The procedure used was straightforward, well defined, proven, and selective. The samples were subjected to isocratic elution using an Agilent Eclipse C18 column (150 × 4.6 mm, 3.5 μ). The mobile phase was supplied at a flow rate of 1.0 mL per minute in a 30 : 70 v/v ratio, containing 0.1% formic acid and acetonitrile. A linear response was observed within the 0.0-150 μg mL concentration range. It was found that the limits of quantitation and detection for pirtobrutinib were 0.1 and 0.3, respectively. The method was assessed for system suitability, linearity, precision, accuracy, and robustness in accordance with standard ICH guidelines. It was found that the results were within acceptable limits. A variety of stress conditions, such as acids, alkalis, hydrolysis, oxidation, reduction as well as photo- and thermal degradations, were applied to the drug to test the method's efficiency and stability. Acidic, alkaline, peroxide, and reduction conditions showed significant degradation. Degradation products produced during the forced degradation studies were analyzed and characterized using mass spectrometry (MS/MS). Thus, the proposed method can also be used for the quantitation of pirtobrutinib in the presence of its degradation products.
本研究考察了pirtobrutinib的碎片化、降解途径和降解产物，这些内容此前尚未见文献报道。当前工作的主要目标是使用液相色谱-串联质谱法（LC-MS/MS）开发、验证和表征强制降解产物。开发了一种等度高效液相色谱法，用于在219 nm波长下定量测定pirtobrutinib。所采用的方法简单明了、定义明确、经过验证且具有选择性。样品使用安捷伦Eclipse C18柱（150×4.6 mm，3.5μm）进行等度洗脱。流动相以每分钟1.0 mL的流速供应，比例为30 : 70 v/v，含有0.1%甲酸和乙腈。在0.0 - 150μg/mL浓度范围内观察到线性响应。发现pirtobrutinib的定量限和检测限分别为0.1和0.3。按照国际协调会议（ICH）标准指南对该方法进行了系统适用性、线性、精密度、准确度和稳健性评估。结果表明在可接受范围内。对该药物施加了各种应激条件，如酸、碱、水解、氧化、还原以及光降解和热降解，以测试该方法的有效性和稳定性。酸性、碱性、过氧化物和还原条件下显示出显著降解现象。使用质谱（MS/MS）对强制降解研究过程中产生的降解产物进行了分析和表征。因此，所提出的方法也可用于在存在降解产物的情况下定量测定pirtobrutinib。
