HPLC-DAD多波长法同时测定丁氯麻滴眼剂中3种成分的含量

Simultaneous Determination of 3 Components in Dinglyuma Eye Drop by HPLC-DAD at Multi-wavelength

目的：建立同时测定丁氯麻滴眼剂中氯霉素、盐酸麻黄碱、盐酸丁卡因含量的方法。方法：采用高效液相色谱-二极管阵列检测器多波长法。色谱柱为ZORBAX SB-C18,流动相为p H 3.0磷酸盐缓冲溶液-乙腈（梯度洗脱）,柱温为室温,流速为1.0 ml/min,进样量为20μl,检测波长为277 nm（氯霉素）、210 nm（盐酸麻黄碱）、310 nm（盐酸丁卡因）。结果：氯霉素、盐酸麻黄碱、盐酸丁卡因的检测质量浓度线性范围分别为12.56-125.6μg/ml（r=0.999 8）、25.55-255.5μg/ml（r=0.999 9）、25.25-252.5μg/ml（r=0.999 9）;精密度、稳定性、重复性试验的RSD〈2%;回收率分别为98.39%-100.44%（RSD=0.79%,n=9）、100.40-101.74%（RSD=0.66%,n=9）、98.83-100.67%（RSD=0.70%,n=9）。结论：该方法灵敏度高、定量准确、专属性强,可用于丁氯麻滴眼剂的质量控制。

OBJECTIVE： To establish a method for the simultaneous determination of chloramphenicol, ephedrine hydrochlo- ride and tetracaine hydrochloride in Dinglyuma eye drop. METHODS：HPLC-DAD at muliti-wavelength was performed on the col- unto of ZORBAX SB-C18 with mobile phase of pH 3.0 Phosphate buffer-acetonitrile （gradient elution）, column temperature was room temperature, flow rate was 1.0 ml/min, injection volume was 20 μl, and the detection wavelength was 277 nm for chloram- phenicol, 210 um for ephedrine hydrochloride and 310 nm for tetracaine hydrochloride. RESULTS： The linear range was 12.56-125.6 μg/ml for chloramphenicol（r=0.999 8）, 25.55-255.5 μg/ml（r=0.999 9） for ephedrine hydrochloride and 25.25-252.5 μg/ml for tetracaine hydrochloride （r=0.999 9）; RSDs of precision, stability and reproducibility tests were lower than 2%; recov- eries were 98.39-100.44% （RSD=0.79%, n=9）, 100.40-101.74% （RSD=0.66% ,n=9） and 98.83-100.67% （RSD=0.70% ,n=9）. CONCLUSIONS ： The method is sensitive, accurate and specific, and can be used for the quality control of Dinglyuma eye drop.