HPLC法同时测定六味能消丸中8种成分的含量

Simultaneous Determination of 8 Components in Liuwei Nengxiao Pills by HPLC

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Diamonsil C_(18),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10μL。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0.121～3.63μg(r=0.999 9)、0.122～3.66μg(r=0.999 9)、0.219～6.57μg(r=0.999 9)、0.016 4～0.492μg(r=0.999 7)、0.017 3～0.519μg(r=0.999 9)、0.015 3～0.459μg(r=0.999 9)、0.007 2～0.216μg(r=0.999 9)、0.016 2～0.486μg(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 ac- id, 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 μg. RESULTS： The linear ranges of alantolactone, isoalantolactone, gallic acid, emodin, aloe-emodine, rhein, phy- scion,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 pg（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.