UPLC法同时测定柴胡疏肝散水煎液冻干物中8个成分的含量

Simultaneous Determination of 8 Components in Lyophilized Product of Chaihu Shugan San Decoction Based on UPLC

目的:建立柴胡疏肝散水煎液冻干物中8个成分含量同时测定的方法。方法:采用超高效液相色谱法对6批柴胡疏肝散水煎液冻干物样品中8个成分芍药内酯苷、芍药苷、阿魏酸、柚皮苷、橙皮苷、苯甲酰芍药苷、甘草酸、α-香附酮进行含量测定。色谱柱为Phenomenex,流动相为0.1%甲酸水-乙腈梯度洗脱,检测波长为250 nm,柱温为30℃,流速为1 mL/min,进样体积为10μL。结果:芍药内酯苷、芍药苷、阿魏酸、柚皮苷、橙皮苷、苯甲酰芍药苷、甘草酸、α-香附酮的检测质量浓度线性范围分别3.606~8.414、23.988~55.972、1.218~2.842、35.964~83.916、12.009~28.021、1.194~2.786、3.609~8.421、5.294~12.352μg/mL(r=0.999 5~0.999 9),定量限分别为0.206、0.178、0.256、0.168、0.196、0.242、0.268、0.157μg/mL;精密度试验、稳定性试验(12 h)、重复性试验中的RSD均<2%(n=5或6),回收率分别为97.93%、98.18%、96.57%、97.61%、98.51%、97.45%、98.14%、96.91%(RSD均<2%);6批样品中芍药内酯苷、芍药苷、阿魏酸、柚皮苷、橙皮苷、苯甲酰芍药苷、甘草酸的平均含量分别为59.258、429.237、23.173、625.847、200.424、15.048、67.620μg/g,α-香附酮因具有挥发性未检测出。结论:建立的含量测定方法简便,结果稳定、可靠、准确,可为柴胡疏肝散水煎液冻干物的质量控制提供参考。

OBJECTIVE:To establish a method for simultaneous determination of 8 components in the lyophilized product of Chaihu shugan san decoction. METHODS:UPLC method was adopted to determine the contents of albiflorin,paeoniflorin,ferulic acid,naringin,hesperidin,benzoyl paeoniflorin,glycyrrhizic acid and α-cyperone in 6 batches of lyophilized product of Chaihu shugan san decoction. The determination was performed on Phenomenex column with mobile phase consisted of 0.1% formic acid water-acetonitrile at the flow rate of 1 m L/min,the sample volume was 10 μL. The detection wavelength was set at 250 nm,and column temperature was 30 ℃. RESULTS:The linear range of albiflorin,paeoniflorin,ferulic acid,naringin,hesperidin,benzoyl paeoniflorin,glycyrrhizic acid and α-cyperone were 3.606-8.414,23.988-55.972,1.218-2.842,35.964-83.916,12.009-28.021,1.194-2.786,3.609-8.421,5.294-12.352 μg/mL,respectively(r=0.999 5-0.999 9). The limits of quantitation were 0.206,0.178,0.256,0.168,0.196,0.242,0.268,0.157 μg/mL,respectively. RSDs of precision,stability(12 h)and repeatability tests were all lower than 2%(n=5 or 6);the recoveries were 97.93%,98.18%,96.57%,97.61%,98.51%,97.45%,98.14%,96.91%(all RSD<2%). The average contents of albiflorin, paeoniflorin, ferulic acid, naringin, hesperidin, benzoyl guanosine, and glycyrrhizic acid in 6 batches of samples were 59.258,429.237,23.173,625.847,200.424,15.048,67.620 μg/g,respectively. α-cyperone was not detected because of its volatility. CONCLUSIONS:The method is simple,stable,reliable and accurate. It could provide reference for quality control of the lyophilized product of Chaihu shugan san decoction.