HPLC法同时测定蒙药复方协日嘎-4中5种有效成分的含量

Simultaneous Determination of 5 Effective Components in Mongolian Medicine Xieriga-4 by HPLC

目的:建立同时测定蒙药复方协日嘎-4中高良姜素、高良姜素-3-甲醚、双去甲氧基姜黄素、去甲氧基姜黄素、姜黄素含量的方法。方法:采用高效液相色谱法。色谱柱为Elite C18,流动相为0.5%磷酸-乙腈(梯度洗脱),流速为0.6 m L/min,检测波长为235 nm(高良姜素、高良姜素-3-甲醚)、420 nm(双去甲氧基姜黄素、去甲氧基姜黄素、姜黄素),柱温为25℃,进样量为10μL。结果:高良姜素、高良姜素-3-甲醚、双去甲氧基姜黄素、去甲氧基姜黄素、姜黄素检测进样量线性范围分别为0.047 2~0.472 0μg(r=0.999 9)、0.104 0~1.040 0μg(r=0.999 9)、0.030 4~0.304 0μg(r=0.999 9)、0.039 2~0.392 0μg(r=0.999 8)、0.232 0~2.320 0μg(r=0.999 7);定量限分别为11.41、12.73、4.92、5.29、5.39 ng,检测限分别为3.41、3.72、1.46、1.59、1.62 ng;精密度、稳定性、重复性试验的RSD均小于3%;加样回收率分别为97.24%~102.93%(RSD=2.01%,n=6)、95.10%~101.50%(RSD=2.34%,n=6)、95.23%~97.61%(RSD=0.97%,n=6)、97.10%~100.65%(RSD=1.24%,n=6)、98.69%~101.15%(RSD=0.99%,n=6)。结论:该方法简单、准确、重复性好,可用于蒙药复方协日嘎-4中高良姜素、高良姜素-3-甲醚、双去甲氧基姜黄素、去甲氧基姜黄素、姜黄素含量的同时测定。

OBJECTIVE:To establish a method for the simultaneous determination of galangin,galangin-3-methyl ether,bisdemethoxycurcumin,demethoxycurcumin and curcumin in mongolian medicine Xieriga-4.METHODS:HPLC method was adopted.The determination was performed on Elite C18 column with mobile phase consisted of 0.5%phosphoric acid-acetonitrile(gradient elution)at the flow rate of 0.6 mL/min.The detection wavelengths were set at 235 nm(galangin,galangin-3-methyl ether)and 420 nm(bisdemethoxycurcumin,demethoxycurcumin and curcumin).The column temperature was set at 25℃,and sample size was 10μL.RESULTS:The linear range of galangin,galangin-3-methyl ether,bisdemethoxycurcumin,demethoxycurcumin and curcumin were 0.047 2-0.472 0μg(r=0.999 9),0.104 0-1.040 0μg(r=0.999 9),0.030 4-0.304 0μg(r=0.999 9),0.039 2-0.392 0μg(r=0.999 8),0.232 0-2.320 0μg(r=0.999 7),respectively.The limits of quantitation were 11.41,12.73,4.92,5.29,5.39 ng;the limits of detection were 3.41,3.72,1.46,1.59,1.62 ng,respectively,respectively.RSDs of precision,stability and repeatability tests were all lower than 3%.The recovery rates were 97.24%-102.93%(RSD=2.01%,n=6),95.10%-101.50%(RSD=2.34%,n=6),95.23%-97.61%(RSD=0.97%,n=6),97.10%-100.65%(RSD=1.24%,n=6),98.69%-101.15%(RSD=0.99%,n=6),respectively.CONCLUSIONS:The method is simple,accurate and reproducible.It can be used for simultaneous determination of galangin,galangin-3-methyl ether,bisdemethoxycurcumin,demethoxycurcumin and curcumin in mongolian medicine Xieriga-4.