HPLC法同时测定六味能消丸中8种成分的含量

摘要

目的:建立同时测定六味能消丸中土木香内酯、异土木香内酯、没食子酸、大黄素、芦荟大黄素、大黄酸、大黄素甲醚、大黄酚含量的方法.方法:采用高效液相色谱法.色谱柱为DiamonsilC18,流动相为甲醇-乙腈-0.1％冰醋酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长为254 nm(土木香内酯、异土木香内酯、大黄素、芦荟大黄素、大黄酸、大黄素甲醚、大黄酚)、270 nm(没食子酸),柱温为25℃,进样量为10 μL.结果:土木香内酯、异土木香内酯、没食子酸、芦荟大黄素、大黄酸、大黄素、大黄素甲醚、大黄酚检测进样量线性范围分别为0.121～3.63 μg(r=0.999 9)、0.122～3.66μg(r=0.999 9)、0.219～6.57 μg(r=0.999 9)、0.016 4～0.492 μg(r=0.999 7)、0.017 3～0.519 μg(r=0.999 9)、0.015 3～0.459 μg(r=0.999 9)、0.007 2～0.216 μg(r=0.999 9)、0.016 2～0.486 μg(r=0.999 9);定量限分别为0.41、0.26、0.35、0.13、0.17、0.14、0.15、0.13 ng,检测限分别为0.12、0.08、0.11、0.04、0.05、0.04、0.05、0.04 ng;精密度、稳定性、重复性试验的RSD＜2.0％;加样回收率分别为98.05％～102.46％(RSD=1.75％,n=6)、98.55％～102.89％ (RSD=1.91％,n=6)、98.53％～102.34％(RSD=1.66％,n=6)、101.71％～103.41％(RSD=0.57％,n=6)、101.04％～103.01％ (RSD=0.69％,n=6)、101.63％～102.75％ (RSD=0.39％,n=6)、96.94％～101.11％ (RSD=1.61％,n=6)、98.06％～99.10％(RSD=0.40％,n=6).结论:该方法准确、简便,可用于六味能消丸中8种成分含量的同时测定.%OBJECTIVE:To establish the method for simultaneous determination of alantolactone,isoalantolactone,gallic acid,emodin,aloe-emodine,rhein,physcion and chrysophanol in Liuwei nengxiao pills.METHODS:HPLC method was adopted.The determination was performed on Diamonsil C18 with mobile phase consisted of methanol-acetonitrile-0.1％ glacial acetic acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelengths were set at 254 nm (alantolactone,isoalantolactone,emodin,aloe-emodine,rhein,physcion and chrysophanol),270 nm (gallic acid).The column temperature was 25 ℃,and sample size was 10 μ L.RESULTS:The linear ranges of alantolactone,isoalantolactone,gallic acid,emodin,aloe-emodine,rhein,physcion,chrysosphanol were 0.121-3.63 μg(r=0.999 9),0.122-3.66 μg(r=0.999 9),0.219-6.57 μg(r=0.999 9),0.016 4-0.492 μg(r=0.999 7),0.017 3-0.519 μg(r=0.999 9),0.015 3-0.459 μg(r=0.999 9),0.007 2-0.216 μg(r=0.999 9),0.016 2-0.486(r=0.999 9).The limits of quantification were 0.41,0.26,0.35,0.13,0.17,0.14,0.15,0.13 ng;limits of detection were 0.12,0.08,0.11,0.04,0.05,0.04,0.05,0.04 ng.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 98.05％-102.46％ (RSD=1.75 ％,n=6),98.55％-102.89％ (RSD=1.91％,n=6),98.53 ％-102.34％ (RSD=1.66％,n=6),101.71％-103.41％ (RSD =0.57 ％,n=6),101.04％-103.01％ (RSD=0.69％,n=6),101.63％-102.75％ (RSD=0.39 ％,n=6),96.94％-101.11％ (RSD=1.61％,n=6),98.06％-99.10％ (RSD=0.40％,n=6).CONCLUSIONS:The method is accurate,simple and suitable for simultaneous determination of 8 components in Liuwei nengxiao pills.