Merey Hanan A, El-Mosallamy Sally S, Hassan Nagiba Y, El-Zeany Badr A
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El Aini Post, Cairo, Egypt.
J Chromatogr Sci. 2019 Apr 1;57(4):305-311. doi: 10.1093/chromsci/bmy112.
Two chromatographic methods were developed, optimized and validated for simultaneous determination of calcipotriol monohydrate (CPM) and betamethasone dipropionate (BMD) in the presence of two dosage form additives named; butylated hydroxytoluene (BHT) and alpha-tocopherol (TOCO). The proposed methods were accurate, sensitive and specific. The first method based on using aluminum thin-layer chromatographic plates precoated with silica gel GF254 as a stationary phase and chloroform-ethyl acetate-toluene (5:5:3, by volume) as a developing system. This was followed by densitometric measurement of the separated bands at 264 nm. Whereas the second method is RP-HPLC where OnyxMonolithic C18® column was used with a gradient profile using methanol, water and acetic acid at flow rate 2.0 mL min-1. Detection was carried out at 264 nm. The methods were validated according to ICH guidelines. The specificity of the developed methods was investigated by analyzing the pharmaceutical dosage form. The validity of the proposed methods was assessed using the standard addition technique. The obtained results were statistically compared with those obtained by the official methods, showing no significant difference with respect to accuracy and precision at P = 0.05.
开发、优化并验证了两种色谱方法，用于在存在两种剂型添加剂（即丁基化羟基甲苯（BHT）和α-生育酚（TOCO））的情况下同时测定一水合钙泊三醇（CPM）和二丙酸倍他米松（BMD）。所提出的方法准确、灵敏且特异。第一种方法基于使用预涂硅胶GF254的铝制薄层色谱板作为固定相，以氯仿 - 乙酸乙酯 - 甲苯（体积比5:5:3）作为展开系统。然后在264nm处对分离的谱带进行光密度测定。而第二种方法是反相高效液相色谱法，使用OnyxMonolithic C18®柱，采用甲醇、水和乙酸的梯度洗脱，流速为2.0 mL min-1。检测在264nm处进行。这些方法根据国际人用药品注册技术协调会（ICH）指南进行了验证。通过分析药物剂型来研究所开发方法的特异性。使用标准加入技术评估所提出方法的有效性。将获得的结果与官方方法获得的结果进行统计学比较，在P = 0.05时，准确度和精密度方面无显著差异。
