HPLC法测定复方水杨酸苯甲酸搽剂有关物质和含量

目的:建立HPLC法测定复方水杨酸苯甲酸搽剂中的有关物质及水杨酸、苯甲酸的含量。方法:采用Agilent Zorbax C_(18)(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇⁃水⁃冰醋酸(40∶60∶1.5),流速为1.0 ml·ml^(-1),检测波长为270 nm,柱温为35℃,进样体积20μl。结果:有关物质方法建立时,水杨酸的3个已知杂质4⁃羟基苯甲酸、4⁃羟基间苯二甲酸、苯酚之间的分离度良好,上述3个杂质的质量浓度分别在3.06~7.14、1.51~3.78、1.63~4.07μg·ml^(-1)范围内线性关系良好,相关系数r均超过0.9995,校正因子分别为0.066,0.11,0.25;平均回收率分别为100.2%,100.4%,100.6%,RSD分别为0.52%,0.61%和1.03%(n=9)。在含量测定方法建立时,水杨酸和苯甲酸的分离良好,2个成分的质量浓度分别在0.3051~0.7119,0.6357~1.4833 mg·ml^(-1)范围内线性关系良好,r均为0.9999,水杨酸和苯甲酸的平均回收率分别为101.5%和101.3%(RSD=0.40%和0.33%,n=9)。4批复方水杨酸苯甲酸搽剂中检出的4⁃羟基苯甲酸、4⁃羟基间苯二甲酸、苯酚杂质量分别为0.010%~0.018%,0.012%~0.014%,0.016%~0.028%;水杨酸和苯甲酸的含量分别为87.0%~90.1%,90.7%~95.2%。结论:本文建立的HPLC法操作简便,灵敏度、稳定性和重复性良好,可用于复方水杨酸苯甲酸搽剂中4⁃羟基苯甲酸、4⁃羟基间苯二甲酸和苯酚等已知杂质的检查,并能同时对水杨酸和苯甲酸两种主成分的含量进行测定。

Chemical Constituents from Piper wallichii

Aim To investigate the chemical constituents of Piper wallichii. Methods Five compounds were isolated by silica gel column chromatography and Sephadex LH-20 gel column chromatography, and the structures of compounds were identified by spectral analysis. Result Five compounds were identified as piperlonguminine （trans, trans） （1）, 4-hydroxy-3,5-dimethoxy-benzoic acid （2）, galgravin （3）, β-sitosterol （4）, and daucosterol （5）. Conclusion Five compounds were isolated from Piper wallichii for the first time, and compounds 1 -3 were isolated from this genus far the first time.