Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr el Aini Street, 11562, Cairo, Egypt.
Chemistry Department, Faculty of Pharmacy, October 6 University, 6 October City, 12585, Giza, Egypt.
J Chromatogr Sci. 2021 Oct 29;59(10):956-963. doi: 10.1093/chromsci/bmab050.
Two stability-indicating chromatographic methods have been established and validated for concurrent determination of probenecid (PRO), colchicine (COL) along with the degradation product of colchicine (COL deg). PRO and COL were exposed to a stress stability study, which includes acidic, alkaline, oxidative, photolytic and thermal degradations. Chromatographic methods included the use of thin layer chromatography (TLC-densitometry) and high performance liquid chromatography (HPLC). In the first method, separation was achieved by using aluminum TLC plates that were precoated with silica gel G.F254 as the stationary phase and ethyl acetate-methanol-33%ammonia (8:1:1, by volume) as a mobile phase. The obtained chromatograms were scanned at 254 nm. The second method was based on HPLC using a RP- C18 column with isocratic elution. Good separation was obtained through a mobile phase comprised of phosphate buffer pH 5-acetonitrile (70:30, v/v) at a flow rate of 1.0 mL min-1 and ultraviolet detection at 254 nm. Different parameters affecting efficiency of the two methods were studied accurately for optimum separation of the three cited components. The suggested methods were validated according to the International Conference on Harmonization (ICH) guidelines and were applied for bulk powder and commercial tablets.
已经建立并验证了两种稳定指示色谱方法，用于同时测定丙磺舒（PRO）、秋水仙碱（COL）以及秋水仙碱的降解产物（COL deg）。PRO 和 COL 进行了稳定性研究，包括酸、碱、氧化、光解和热降解。色谱方法包括使用薄层色谱（TLC- 密度法）和高效液相色谱（HPLC）。在第一种方法中，使用预先涂有硅胶 G.F254 的铝薄层色谱板作为固定相，乙酸乙酯-甲醇-33%氨（8:1:1，体积比）作为流动相实现分离。获得的色谱图在 254nm 处扫描。第二种方法基于使用 RP-C18 柱的 HPLC，采用等度洗脱。通过由磷酸盐缓冲液 pH 5-乙腈（70:30，v/v）组成的流动相，在 1.0mL min-1 的流速下和在 254nm 处的紫外检测，获得良好的分离。准确研究了影响两种方法效率的不同参数，以实现三种引用成分的最佳分离。根据国际协调会议（ICH）指南验证了建议的方法，并将其应用于散装粉末和商业片剂。
