Modern College of Pharmacy, Nigdi, QAT, Pune, Maharashtra 412109, India.
Callidus Research Laboratories Pvt. Ltd, PAP-A-29/1, Chakan Industrial Area Phase-IV, Nighoje, Tal-Khed, Pune, Maharashtra 410501, India.
J Chromatogr Sci. 2021 May 20;59(6):555-565. doi: 10.1093/chromsci/bmaa125.
A novel, delicate, stability-indicating, gradient, reversed-phase high-performance liquid chromatographic method has been established for the quantitative estimation of methocarbamol (MTC) and its impurities present in a pharmaceutical oral suspension. XBridge C18, 5 μm, 250 mm × 4.6 mm column was used to accomplish chromatographic separation with a buffered mobile phase consisting of a mixture of 0.01 M of sodium dihydrogen phosphate (pH 7.0 buffer) and methanol in the ratio of 95:05 (v/v), respectively, were used as solvent A and a mixture of methanol and Milli-Q water in the ratio 90:10 (v/v), respectively, was used as solvent B. Analysis was carried out at 0.8 mL/min flow rate and the detection wavelength at 225 nm. The compartment temperature of the column is put at 25°C. The resolution of MTC and its four impurities has been attained >2.0 for all pairs of compounds. Significant degradation of MTC was photolytic, thermal and oxidative stress conditions. Validation of the developed method was performed as stated by the International Conference on Harmonization guidelines with regard to all validation parameters like specificity, accuracy, linearity, precision, limit of detection, limit of quantitation and robustness.
建立了一种新颖、灵敏、专属性强、梯度洗脱、反相高效液相色谱法,用于定量测定药物口服混悬剂中甲氨蝶呤(MTC)及其杂质。采用 XBridge C18,5μm,250mm×4.6mm 柱,以含有 0.01M 磷酸二氢钠(pH7.0 缓冲液)和甲醇的混合溶液(体积比 95:05)为溶剂 A,以甲醇和 Milli-Q 水的混合溶液(体积比 90:10)为溶剂 B,分别作为流动相,完成色谱分离。分析流速为 0.8mL/min,检测波长为 225nm。柱温为 25°C。MTC 与其四种杂质的分离度均大于 2.0。MTC 显著降解发生在光解、热和氧化应激条件下。所建立的方法按照国际协调会议指南进行了验证,包括专属性、准确度、线性、精密度、检测限、定量限和耐用性等所有验证参数。