HPLC切换波长法同时测定桑菊消肿贴膏中7种成分含量

Simultaneous Determination of Seven Components in Sangju Xiaozhong Gel Plaster by HPLC Wavelength Switching Method

目的采用反相高效液相色谱切换波长法建立同时测定桑菊消肿贴膏中绿原酸、芍药苷、阿魏酸、芦丁、异槲皮苷、木犀草苷和丹皮酚7种成分含量的方法。方法采用WondaCract ODS-2色谱柱(4.6 mm×250 mm,5μm);以乙腈-0.5%磷酸水+0.1 moL/L磷酸二氢钠溶液为流动相,梯度洗脱,流速0.8 mL/min,柱温40℃,检测波长分别为210 nm(绿原酸)、230 nm(芍药苷)、325 nm(阿魏酸)、344 nm(芦丁、异槲皮苷)、351 nm(木犀草苷)、275 nm(丹皮酚)。结果绿原酸、芍药苷、阿魏酸、芦丁、异槲皮苷、木犀草苷和丹皮酚的质量浓度分别在0.0578~2.8880μg(r=0.9999)、0.0964~4.8200μg(r=0.9998)、0.0282~1.4100μg(r=0.9995)、0.0458~2.2840μg(r=0.9995)、0.0526~2.6330μg(r=0.9996)、0.0154~0.7680μg(r=0.9996)、0.0306~1.5260μg(r=0.9995)范围内与峰面积呈良好的线性关系,仪器精密度及样品稳定性均良好,平均回收率依次为96.29%(RSD=1.84%)、97.23%(RSD=1.02%)、97.72%(RSD=1.82%)、100.47%(RSD=1.27%)、100.10%(RSD=1.26%)、99.40%(RSD=2.90%)、99.87%(RSD=2.98%)。结论本研究建立的测定方法准确可靠、操作简便、重复性好,可用于桑菊消肿贴膏中7种成分的含量测定。

Objective To establish a wavelength switching method for simultaneous determination of chlorogenic acid,paeoniflorin,ferulic acid,rutin,isoquercitrin,luteoloside,and paeonol in Sangju Xiaozhong Gel Plaster.Methods The WondaCract ODS-2 chromatographic column(4.6 mm×250 mm,5μm)was used for the separation,with gradient elution.The mobile phase was acetonitrile and 0.5%phosphoric acid solution with 0.1 moL/L sodium dihydrogen phosphate at a flow rate of 0.8 mL/min.The column temperature was 40℃.The detection wavelength was set at 210 nm(chlorogenic acid),230 nm(paeoniflorin),325 nm(ferulic acid),344 nm(rutin and isoquercitrin),351 nm(luteoloside),and 275 nm(paeonol),respectively.Results The method achieved a good linearity in the liner ranges of 0.0578‒2.8880μg for chlorogenic acid(r=0.9999),0.0964‒4.8200μg for paeoniflorin(r=0.9998),0.0282‒1.4100μg for ferulic acid(r=0.9995),0.0458‒2.2840μg for rutin(r=0.9995),0.0526‒2.6330μg for isoquercitrin(r=0.9996),0.0154‒0.7680μg for luteoloside(r=0.9996),and 0.0306‒1.5260μg for paeonol(r=0.9995).Degrees of precision of instrument and the stability of samples were good.The average recovery rates were 96.29%(RSD=1.84%),97.23%(RSD=1.02%),97.72%(RSD=1.82%),100.47%(RSD=1.27%),100.10%(RSD=1.26%),99.40%(RSD=2.90%),and 99.87%(RSD=2.98%),respectively.Conclusion The method established in this study is accurate,convenient and reproducible,which can be used for the content determination of seven components in Sangju Xiaozhong Gel Plaster.