HPLC法测定不同贮藏条件下川芎中10种化学成分的含量

目的:建立同时测定不同贮藏条件下川芎中绿原酸、川芎嗪、阿魏酸、洋川芎内酯I、洋川芎内酯H、阿魏酸松柏酯、洋川芎内酯A、正丁基苯酞、Z-藁本内酯、丁烯基苯酞含量的方法。方法:采用高效液相色谱法。色谱柱为Boston C₁₈,流动相为乙腈-0.5%醋酸溶液(梯度洗脱),流速为1.0 m L/min,检测波长为285 nm,柱温为30℃,进样量为5μL。结果:绿原酸、川芎嗪、阿魏酸、洋川芎内酯I、洋川芎内酯H、阿魏酸松柏酯、洋川芎内酯A、正丁基苯酞、Z-藁本内酯、丁烯基苯酞检测进样量线性范围分别为0.030 53～0.519 01μg(r=0.999 5)、0.001 02～0.017 34μg(r=0.999 9)、0.012 83～0.218 11μg(r=0.999 5)、0.007 63～0.129 71μg(r=0.999 7)、0.001 76～0.029 92μg(r=0.999 5)、0.054 74～0.930 58μg(r=0.999 9)、0.215 80～3.668 60μg(r=0.999 9)、0.018 02～0.306 34μg(r=0.999 7)、0.232 50～3.952 50μg(r=0.999 9)、0.002 40～0.040 80μg(r=0.999 5);定量限分别为2.073 7、0.556 6、0.753 8、0.231 5、0.306 9、0.925 2、2.295 3、4.624 0、3.215 3、0.910 5 ng,检测限分别为0.622 1、0.167 0、0.226 1、0.069 4、0.092 1、0.277 6、0.688 6、1.387 2、0.964 6、0.273 1 ng;精密度、稳定性、重复性试验的RSD均小于5%(n=6);加样回收率分别为95.90%～103.28%(RSD=2.99%,n=6)、88.24%～107.84%(RSD=4.89%,n=6)、95.06%～102.08%(RSD=3.97%,n=6)、93.67%～101.05%(RSD=1.02%,n=6)、94.81%～104.33%(RSD=2.34%,n=6)、94.41%～105.59%(RSD=4.32%,n=6)、92.76%～104.83%(RSD=1.95%,n=6)、87.22%～102.56%(RSD=2.89%,n=6)、94.04%～99.52%(RSD=0.92%,n=6)、88.51%～103.83%(RSD=4.89%,n=6)。5、15℃贮藏条件下,药材药品未见明显变质;室温条件下,部分样品出现虫蛀、霉变;6批药材样品各成分含量分别为0.047 7%～0.160 8%、0.006 1%～0.022 7%、0.048 2%～0.172 2%、0.023 3%～0.145 2%、0.004 6%～0.030 7%、0.085 2%～0.835 4%、0.182 6%～2.112 7%、0.009 9%～0.098 3%、0.614 9%～3.176 2%、0.005 7～0.036 9%,均呈下降趋势,且下降速率为5℃<15℃<室温;在相同贮藏温度下的下降速率为聚乙烯塑料袋<聚丙烯编织袋。结论:该方法准确、可行,可用于同时测定川芎中10种成分的含量。建议川芎药材密封包装后于干燥、阴凉处贮藏,且贮藏时间不宜过长。

Content Determination of 10 Kinds of Chemical Components in Ligusticum chuanxiong under Different Storage Conditions by HPLC

OBJECTIVE:To establish the method for the content determination of chlorogenic acid,ligustrazine,ferulic acid,senkyunolide I,senkyunolide H,coniferly ferulate,senkyunolide A,n-butylphtalide,Z-ligustilide and n-butylidenephthalide in Ligusticum chuanxiong under different storage conditions.METHODS:HPLC method was adopted.The determination was performed on Boston C18 column with mobile phase consisted of acetonitrile-0.5%acetic acid solution(gradient elution)at the flow rate of 1.0 mL/min.The detection wavelength was set at 285 nm,and the column temperature was 30℃.The sample size was 5μL.RESULTS:The linear range of chlorogenic acid,ligustrazine,ferulic acid,senkyunolide I,senkyunolide H,coniferly ferulate,senkyunolide A,n-butylphtalide,Z-ligustilide n-butylidenephthalide were 0.030 53-0.519 01μg(r=0.999 5),0.001 02-0.017 34μg(r=0.999 9),0.012 83-0.218 11μg(r=0.999 5),0.007 63-0.129 71μg(r=0.999 7),0.001 76-0.029 92μg(r=0.999 5),0.054 74-0.930 58μg(r=0.999 9),0.215 80-3.668 60μg(r=0.999 9),0.018 02-0.306 34μg(r=0.999 7),0.232 50-3.952 50μg(r=0.999 9),0.002 40-0.040 80μg(r=0.999 5).The limits of quantitation were 2.073 7,0.556 6,0.753 8,0.231 5,0.306 9,0.925 2,2.295 3,4.624 0,3.215 3,0.910 5ng,respectively.The detection limits were 0.622 1,0.167 0,0.226 1,0.069 4,0.092 1,0.277 6,0.688 6,1.387 2,0.964 6,0.273 1 ng,respectively.RSD of precision,stability and repeatability tests were all less than 5%(n=6).The recovery rates were 95.90%-103.28%(RSD=2.99%,n=6),88.24%-107.84%(RSD=4.89%,n=6),95.06%-102.08%(RSD=3.97%,n=6),93.67%-101.05%(RSD=1.02%,n=6),94.81%-104.33%(RSD=2.34%,n=6),94.41%-105.59%(RSD=4.32%,n=6),92.76%-104.83%(RSD=1.95%,n=6),87.22%-102.56%(RSD=2.89%,n=6),94.04%-99.52%(RSD=0.92%,n=6),88.51%-103.83%(RSD=4.89%,n=6),respectively.At 5℃and 15℃,no obvious deterioration was observed in medicinal materials.At room temperature,some samples were moth-eaten and mildewed.The content ranges of 6 batches of samples were 0.047 7%-0.160 8%,0.006 1%-0.022 7%,0.048 2%-0.172 2%,0.023 3%-0.145 2%,0.004 6%-0.030 7%,0.085 2%-0.835 4%,0.182 6%-2.112 7%,0.009 9%-0.098 3%,0.614 9%-3.176 2%and 0.005 7%-0.036 9%,showing decreasing trend;the decrease rate was in descending order 5℃<15℃