Parab Manali, Shirsat Vaishali A, Kodgule Yogita M, Kodgule Mandar
Bombay College of Pharmacy (Autonomous), Kalina, Santacruz East, Mumbai 400098, India.
IQGEN-X Pharma Pvt. Ltd., A-165, Khairane Road, Sector 2, Kopar Khairane, Navi Mumbai 400710, India.
Int J Anal Chem. 2021 May 11;2021:5570173. doi: 10.1155/2021/5570173. eCollection 2021.
Neostigmine methylsulfate is an anticholinesterase agent and is clinically used for treating myasthenia gravis. It is also used for reversing nondepolarising neuromuscular blocking agents. Neostigmine methylsulfate may be administered by intravenous, intramuscular, or subcutaneous injection. In this research paper, a distinct stability-indicating reverse phase HPLC method was developed and validated for the quantitative determination of related impurities and degradation impurities in neostigmine methylsulfate API and injection formulation. The specific objective was to improve the resolution between European Pharmacopoeia listed impurity A and impurity B and degradation impurity of neostigmine methylsulfate API and injection formulation. The analysis was performed using Kromasil C column at 30°C of column oven temperature with phosphate-buffer/acetonitrile in a gradient mode. The RP-HPLC method was developed and validated for in-house neostigmine methylsulfate synthesis process sample and injection formulation. The injection formulation sample was studied for accelerated stability, temperature cycling stability, and photostability. The validation studies for neostigmine methylsulfate synthesis process API were studied using impurity A, impurity B, and impurity C. The analytical method validation parameters studied were specificity, precision, linearity, limit of detection, limit of quantitation, accuracy, and robustness. The API and the injection formulation were subjected to forced degradation under acid, alkali, oxidation, and photolytic and thermal conditions. The proposed method showed a significantly improved RRT (Relative Retention Time) of impurity A and impurity B with a resolution greater than 1.5. The developed method eliminates the use of an ion-pairing agent and thereby a good performance of column was established.
甲基硫酸新斯的明是一种抗胆碱酯酶药，临床上用于治疗重症肌无力。它还用于逆转非去极化型神经肌肉阻滞剂的作用。甲基硫酸新斯的明可通过静脉注射、肌肉注射或皮下注射给药。在本研究论文中，开发并验证了一种独特的稳定性指示反相高效液相色谱法，用于定量测定甲基硫酸新斯的明原料药及注射剂中的有关物质和降解杂质。具体目标是提高欧洲药典列出的杂质A与杂质B以及甲基硫酸新斯的明原料药及注射剂降解杂质之间的分离度。分析在柱温30℃的Kromasil C柱上进行，以磷酸盐缓冲液/乙腈为流动相，采用梯度洗脱模式。该反相高效液相色谱法针对内部甲基硫酸新斯的明合成工艺样品及注射剂进行了开发和验证。对注射剂样品进行了加速稳定性、温度循环稳定性和光稳定性研究。对甲基硫酸新斯的明合成工艺原料药的验证研究使用了杂质A、杂质B和杂质C。所研究的分析方法验证参数包括专属性、精密度、线性、检测限、定量限、准确度和耐用性。原料药和注射剂在酸、碱、氧化、光解和热条件下进行强制降解。所提出的方法显示杂质A和杂质B的相对保留时间（RRT）显著提高，分离度大于1.5。所开发的方法无需使用离子对试剂，从而建立了良好的色谱柱性能。
