HPLC法同时测定通舒口爽片中9个成分的含量

Simultaneous determination of nine constituents in Tongshu Koushuang tablets by HPLC

目的:采用高效液相色谱(HPLC)技术,建立同时测定通舒口爽片(大黄、枳实、秦艽等)中马钱苷酸、龙胆苦苷、橙皮苷、丹皮酚、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚9个成分的含量。方法:样品经甲醇回流提取后,分析采用Thermo Acclaim-C_(18)(250 mm×4.6 mm,5μm)色谱柱,流动相为0.15%磷酸(乙腈,梯度洗脱,流速1.0 mL·min^(-1),柱温30℃,进样量10μL,检测波长为236 nm(检测马钱苷酸)、280 nm(检测龙胆苦苷、橙皮苷、丹皮酚)、254 nm(检测芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚)。结果:马钱苷酸、龙胆苦苷、橙皮苷、丹皮酚、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚在各自线性范围内呈良好的关系(r为0.9991~0.9999),平均加样回收率(RSD)分别为99.6%(RSD=1.9%)、101.1%(RSD=2.0%)、100.5%(RSD=1.5%)、101.6%(RSD=0.51%)、99.8%(RSD=1.4%)、100.9%(RSD=1.4%)、100.8%(RSD=1.8%)、102.7%(RSD=1.2%)、102.9%(RSD=0.86%),6批样品中9个成分的含量范围分别为1.871~3.517 mg·g^(-1)、6.274~12.55 mg·g^(-1)、2.140~3.946 mg·g^(-1)、1.393~2.018 mg·g^(-1)、0.3948~0.8072 mg·g^(-1)、0.6575~1.246 mg·g^(-1)、0.4847~0.9158 mg·g^(-1)、0.8398~1.429 mg·g^(-1)、0.3273~0.5479 mg·g^(-1)。结论:所建立的方法简单准确,可用于通舒口爽片的质量控制与评价。

Objective:To establish an HPLC method for simultaneous determination of loganic acid,gentiopicroside,hesperidin,paeonol,aloe-emodin,rhein,emodin,chrysophanol and physcion in Tongshu Koushuang tablets(Rhei Radix et Rhizoma,Aurantii Fructus Immaturus and Gentianae Macrophyllae Radix,etc.).Methods:The samples were extracted by refluxing with methanol and the separation was performed on a Thermo Acclaim-C_(18)(250 mm×4.6 mm,5μm)column at 30℃,with mobile phase consisting of 0.15%phosphoric acid solution and acetonitrile with a gradient elution at a flow rate of 1.0 mL·min^(-1).The injection volume was 10μL,and the detection wavelength was set at 236 nm(detecting loganic acid),280 nm(detecting gentiopicroside,hesperidin and paeonol)and 254 nm(detecting aloe-emodin,rhein,emodin,chrysophanol and physcion).Results:Nine constituents(loganic acid,gentiopicroside,hesperidin,paeonol,aloe-emodin,rhein,emodin,chrysophanol and physcion)were in good linearity within their own ranges(r were 0.9991-0.9999),and the average recoveries were 99.6%(RSD=1.9%),101.1%(RSD=2.0%),103.0%(RSD=1.5%),101.6%(RSD=0.51%),99.8%(RSD=1.4%),100.9%(RSD=1.4%),100.8%(RSD=1.8%),102.7%(RSD=1.2%)and 102.9%(RSD=0.86%),respectively.The content ranges of nine constituents were 1.871-3.517 mg·g^(-1),6.274-12.55 mg·g^(-1),2.140-3.946 mg·g^(-1),1.393-2.018 mg·g^(-1),0.3948-0.8072 mg·g^(-1),0.6575-1.246 mg·g^(-1),0.4847-0.9158 mg·g^(-1),0.8398-1.429 mg·g^(-1)and 0.3273-0.5479 mg·g^(-1),respectively.Conclusion:This simple and accurate method can be used for the quality control and evaluation of Tongshu Koushuang tablets.