UPLC波长切换法同时测定沉香化气丸中7个成分的含量

Simultaneous determination of seven components in Chenxiang Huaqi pills by UPLC with wavelength switching

目的:建立UPLC波长切换法同时测定沉香化气丸中甘草苷、芸香柚皮苷、柚皮苷、橙皮苷、甘草酸、木香烃内酯和广藿香酮的含量。方法:采用Kinetex C18色谱柱(4.6 mm×100 mm,2.7μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速0.8 mL·min^(-1),波长切换(0~20 min,210 nm,检测甘草苷、芸香柚皮苷、柚皮苷、橙皮苷;20~25 min,254 nm,检测甘草酸;25~30.8 min,210 nm,检测木香烃内酯;30.8~34 min,310 nm,检测广藿香酮),柱温35℃。结果:待测成分峰与相邻峰达到基线分离,甘草苷、芸香柚皮苷、柚皮苷、橙皮苷、甘草酸、木香烃内酯和广藿香酮的质量浓度分别在1.659~39.82μg·mL^(-1)(r=1.000)、1.729~41.50μg·mL^(-1)(r=0.999 8)、0.663 4~15.93μg·mL^(-1)(r=0.999 7)、9.637~240.9μg·mL^(-1)(r=0.999 7)、2.127~51.06μg·mL^(-1)(r=0.999 7)、1.794~43.08μg·mL^(-1)(r=0.999 6)、0.9080~21.80μg·mL^(-1)(r=0.999 4)范围内与峰面积呈良好的线性关系;平均加样回收率(n=6)分别为98.7%(RSD=2.4%)、101.7%(RSD=2.6%)、100.5%(RSD=2.9%)、99.0%(RSD=2.8%)、99.5%(RSD=2.5%)、99.3%(RSD=2.2%)、101.7%(RSD=1.7%);方法重复性(n=6)的RSD均小于2.9%;供试品溶液在室温条件下24 h内稳定,RSD均小于1.8%。8批样品中甘草苷、芸香柚皮苷、柚皮苷、橙皮苷、甘草酸、木香烃内酯和广藿香酮7个成分的含量测定结果分别为0.424~0.989、0.482~0.802、0.105~0.204、3.170~4.842、0.702~0.997、0.183~0.916、0.216~0.385 mg·g^(-1)。结论:该方法为沉香化气丸的质量标准研究提供参考。

Objective：To develop a method for simultaneous determination the contents of liquiritin,narirutin,naringin,hesperidin,glycyrrhizic acid,costunolide and pogostone in Chenxiang Huaqi pills by UPLC with wavelength switching.Methods：The analyses were carried out on a Kinetex C18 column（4.6 mm×100 mm, 2.7 μm）with the mobile phase consisting of acetonitrile and 0.1% phosphoric acid in a gradient mode at a flow rate of 1 mL·min-1.The detection wavelength was set at 210 nm for liquiritin,narirutin,naringin and hesperidin in the first 20 min,254 nm for glycyrrhizic acid during 20-25 min,210 nm for costunolide during 25-30.8 min,310 nm for pogostone during 30.8-34 min.The column temperature was at 35 ℃.Results：Baseline separation of the analytes and their adjacent peaks reaching the standard by chromatography.Good linearity was obtained for liquiritin,narirutin,naringin,hesperidin,glycyrrhizic acid,costunolide and pogostone in ranges of 1.659-39.82 μg·mL-1（r=1.000）,1.729-41.50 μg·mL-1（r=0.999 8）,0.663 4-15.93 μg·mL-1（r=0.999 7）,9.637-240.9 μg·mL-1（r=0.999 7）,2.127-51.06 μg·mL-1（r=0.999 7）,1.794-43.08 μg·mL-1（r=0.999 6）,and 0.9080-21.80 μg·mL-1（r=0.999 4）,respectively.The average recoveries（n=6）of the above seven components were 98.7%（RSD=2.4%）,101.7%（RSD=2.6%）,100.5%（RSD=2.9%）,99.0%（RSD=2.8%）,99.5%（RSD=2.5%）,99.3%（RSD=2.2%）,and 101.7%（RSD=1.7%）,respectively.The repeatability was good with RSD less than 2.9%.The seven analytes were stable at room temperature within 24 h with RSD less than 1.8%.The contents in 8 samples were in the ranges of 0.424-0.989 mg·g-1 for liquiritin,0.482-0.802 mg·g-1 for narirutin,0.105-0.204 mg·g-1 for naringin,3.170-4.842 mg·g-1 for hesperidin,0.702-0.997 mg·g-1 for glycyrrhizic acid,0.183-0.916 mg·g-1 for costunolide and 0.216-0.385 mg·g-1 for pogostone.Conclusion：The method offered reference for quality standard of Chenxiang Huaqi pills.