卡泊三醇有关物质测定方法研究及原料稳定性考察

Study on Determination Method of Related Substances of Calcipotriol and Stability of Raw Materials

目的：建立卡泊三醇原料中有关物质的测定方法，考察卡泊三醇原料的稳定性。方法采用高效液相色谱法，色谱柱为Kromasil 100-5 C18柱，柱温为35℃，流速为1.0 mL·min-1，流动相为乙腈-甲醇-0.01 mol·L-1磷酸盐缓冲液（pH 6.0±0.1）20：50：30，检测波长为264 nm。考察光、热、酸、碱及氧化对卡泊三醇原料的影响。结果样品色谱图中卡泊三醇色谱峰与杂质峰有较好的分离度，且重现性较好，卡泊三醇在0.1385-37.2218μg·mL-1浓度范围内与峰面积呈良好的线性关系，线性方程Y=120000X＋2969.5，R=0.9994，最低定量限为0.14μg·mL-1。破坏性试验结果显示，卡泊三醇对光、热、酸、碱及氧化都比较敏感。结论该法操作简单、准确，灵敏度高，精密度高，重现性好，可用于卡泊三醇原料中有关物质的分离分析。卡泊三醇制剂生产和贮存过程中，应尽量避免光、热、酸、碱以及氧化对产品质量的影响。

Objective To establish the determination methods of related substances and to investigate the stabilitiy of calcipotriol raw materials.Methods High performance liquid chromatography （HPLC） was used for the determination of related substances in calcipotriol. The column was Kromasil 100-5 C18 and the column temperature was 35℃. The mobile phase was acetonitrile- methanol - 0.01 mol·L-1 phosphate buffer solution （pH 6.0 ± 0.1） （20：50：30） with the lfow rate of 1.0 mL·min-1. The detection wavelength was 264 nm. Moreover, the effects of light, heat, acid, alkali and oxidation on calcipotriol were investigated.Results The resolution and reproducibility of the principal component peak and impurity peaks of calcipotriol is good. It showed a good linear relationship in the concentration range of 0.138 5-37.221 8 μg·mL-1, and the linear equation was Y=120 000X＋2 969.5, R=0.999 4. The limit of quantitation of the method was 0.14 μg·mL-1. Destructive test results showed that calcipotriol was sensitive to light, heat, acid, alkali and oxidation.Conclusion The method is simple, accurate, with high sensitivity, high precision and good reproducibility, which can be used for the separation and analysis of the related substances in calcipotriol raw materials. Light, heat, acid, alkali, as well as the impact of oxidation should be avoided in the process of production and storage of the calcipotriol raw materials and its preparations.