Development and Validation of UV-Spectrophotometric and Stability Indicating RP-HPLC Method of Calcipotriene in Bulk Drug and Pharmaceutical Formulation

The objective of current study was to develop and validate stability indicating UV Spectrophotometric and RP-HPLC method for Calcipotriene in bulk drug and pharmaceutical formulation. UV Spectrophotometric method was developed utilizing Analytical Technologies limited. Detection was carried out at absorption maxima at 260nm using methanol as a solvent. Beer's law was followed in the concentration range of 5-25 µg/mL, and the drug was quantitated using A1% 1cm at 260 nm, which produced a correlation coefficient that was less than 1. The Chromatographic separation of analyte was achieved on Agilent C18 (4.6mm x 250mm, 5µm) column with mobile phase consisting of Methanol and 0.1 % Acetic Acid in the ratio of 65:35% v/v at flow rate of 1.0 ml/min. The retention time was found to be 5.273 min. Calcipotriene was subjected to forced degradation studies under different stress conditions like acid hydrolysis, alkaline hydrolysis, and hydrogen peroxide oxidation. The developed method was validated according to the guidelines of International Conference on Harmonization (ICH) for various parameters like linearity, precision, accuracy, robustness, limit of detection and limit of quantitation. The findings showed that the method performed well in accordance with the standards. The proposed method is simple, accurate, precise, economic, reproducible and stability indicating and hence suitable for routine quality control analysis of Calcipotriene in bulk drug as well as in formulations.本研究旨在开发并验证一种可用于卡泊三醇（Calcipotriene）原料药及药物制剂的稳定性指示型紫外分光光度法与反相高效液相色谱法（RP-HPLC）。紫外分光光度法依托分析技术有限责任公司（Analytical Technologies Limited）的分析技术完成开发，以甲醇为溶剂，在260nm的最大吸收波长处进行检测。该方法在5~25 μg/mL的浓度范围内符合比尔定律，采用260nm处的百分吸光系数（A1%1cm）对药物进行定量，所得相关系数小于1。分析物的色谱分离采用安捷伦C18色谱柱（4.6mm×250mm，5μm）完成，流动相为体积比65:35的甲醇与0.1%乙酸溶液，流速为1.0 mL/min，待测物的保留时间为5.273 min。对卡泊三醇开展了不同胁迫条件下的强制降解研究，涵盖酸水解、碱水解及过氧化氢氧化。依据人用药品注册技术要求国际协调会（ICH）的指导原则，对所建立的方法进行了线性、精密度、准确度、耐用性、检测限与定量限等多项参数的验证。研究结果表明，该方法完全符合各项标准要求。所开发的方法具备简便、准确、精密、经济、可重现且具有稳定性指示性的优势，因此适用于卡泊三醇原料药及其制剂的常规质量控制分析。