[关键词]
[摘要]
目的 采用HPLC梯度洗脱法同时测定沉香化滞丸中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚11种成分。方法 采用Thermo Syncronis C18色谱柱(250 mm×4.6 mm,5 μm),流动相为水-乙腈,梯度洗脱:0~10 min,20%乙腈;10~20 min,20%~40%乙腈;20~24 min,40%乙腈;24~26 min,40%~52%乙腈;26~30 min,52%乙腈;30~31 min,52%~90%乙腈;31~35 min,90%乙腈;35~40 min,90%~100%乙腈;40~43 min,100%乙腈;43~45 min,100%~20%乙腈;检测波长215 nm,体积流量1.0 mL/min,柱温30℃,进样量20 μL。结果 各成分在43 min内分离良好,沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚的线性范围分别为1.4~13.6、10.0~200.0、31.5~315.0、1.0~120.1、1.8~50.6、0.93~10.1、1.8~30.0、0.2~40.3、1.8~18.1、1.7~25.0、0.45~10.70μg/mL;样品中各成分的平均回收率均在98.90%~100.87%;11种成分精密度RSD在0.55%~1.54%;供试品溶液在30 h内稳定性良好,RSD在0.75%~1.94%;重复性RSD在0.39%~1.73%。6批次样品中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚质量分数分别为92.0~201.0、511.5~9 033.0、5 475.0~12 635.5、54.5~5 095.5、192.0~2 137.5、117.0~391.5、106.5~1 281.5、13.0~136.5、93.5~199.0、177.0~1 207.0、33.5~251.5 μg/g。结论 本方法准确、快速、简便,重复性好,精密度高,适用于沉香化滞丸中多种活性成分的定量分析。
[Key word]
[Abstract]
Objective To develop an HPLC method for simultaneous determination of the eleven constituents (agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion) in Chenxiang Huazhi Pills (CHP) by HPLC with gradient elution. Methods The chromatographic separation was performed on an Thermo Syncronis C18 column (4.6 mm×250 mm, 5 μm) which was operated at 30℃. The mobile phase was a linear gradient prepared from water (A) and acetonitrile (B). The linear gradient elution program was programmed as follows:0-10 min, 20% acetonitrile; 10-20 min, 20%-40% acetonitrile; 20-24 min, 40% acetonitrile; 24-26 min, 40%-52% acetonitrile; 26-30 min, 52% acetonitrile; 30-31 min, 52%-90% acetonitrile; 31-35 min, 90% acetonitrile; 35-40 min, 90%-100% acetonitrile; 40-43 min, 100% acetonitrile; 43-45 min, 100%-20% acetonitrile. The flow rate was 1 mL/min and the detection wavelength was 215 nm. Results The analysis permitted very good separation of eleven constituents within 43 min. A good linear relationship between the peak area and the injection volume was obtained. The ranges of the eleven constituents were 1.4-13.6, 10.0-200.0, 31.5-315.0, 1.0-120.1, 1.8-50.6, 0.93-10.1, 1.8-30.0, 0.2-40.3, 1.8-18.1, 1.7-25.0, and 0.45-10.70 μg/mL. The average recoveries of eleven constituents in the samples were in the range of 98.90%-100.87%. The precision RSD of the peak areas of the 11 components ranged from 0.55%-1.54%; Eleven components had good stability within 30 h, and the concentration RSD of each component ranged from 0.75% to 1.94%; The repeatability RSD of each component ranged from 0.39% to 1.73%. The content of agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion in six batches were 92.0-201.0, 511.5-9 033.0, 5 475.0-12 635.5, 54.5-5 095.5, 192.0-2 137.5, 117.0-391.5, 106.5-1 281.5, 13.0-136.5, 93.5-199.0, 177.0-1 207.0, and 33.5-251.5 μg/g, respectively. Conclusion The method is accurate, rapid and simple with high sensitivity, precision and repeatability, which has been successfully applied as an effective tool for the multicomponent analysis of CHP.
[中图分类号]
R927.2
[基金项目]
国家级大学生创新创业训练计划项目(201410316060);大学生创新药物研制能力提高项目(J1030830)