HPLC双波长梯度洗脱法同时测定退热解毒注射液中绿原酸、连翘苷、丹皮酚的含量

Content Determination of Chlorogenic Acid,Forsythin and Paeonol in Tuire Jiedu Injection by Dual Wavelength HPLC with Gradient Elution

目的：建立同时测定退热解毒注射液中绿原酸、连翘苷、丹皮酚含量的HPLC双渡长梯度洗脱法。方法：色谱柱为Kromasil C18（4．6mm×250mm，5μm）；以乙腈-水（25：75）为流动相A，0．3％磷酸为流动相B，梯度洗脱。检测波长λ1=327nm，A2=276nm；流速为1．0mL·min-1；柱温为30℃。结果：绿原酸、连翘苷、丹皮酚分别在8．34～66．69、8．01。64．08、4．01～32．05μg·mL-1范围内线性关系良好（r=0．9999）。平均回收率分别为99．14％、99．01％、99．00％。相应的RSD分别为0．72％、0．47％、0．82％（n=6）。结论：方法简便可靠，结果重现性好，为控制退热解毒注射液的内在质量提供了科学依据。

Objective： To establish a method for content determination of chlorogenic acid and forsythin and paeonol in Tuire Jiedu Injection by dual wavelength HPLC with gradient elution. Method：The Kromasil C18 column （4.6 mm ×250 mm,5 μm） was used. The mobile phase was consisted of A ： aeetonitrile-water （ 25： 75 ） and B ：0. 3 % phosphoric acid for gradient elution. The detection wavelength was at λ1 =327 nm,λ2 =276 nm , the flow rate was 1.0 mL· min-1. The temperature of the column at 30 ℃. Result： the linear range for chlorogenic acid and forsythin and paeonol was 8. 34 - 66. 69, 8. 01 - 64. 08, 4. 01 - 32. 05μg · mL-1 （ r =0. 999 9 ） , the average recovery was 99.14% （ RSD = 0.72% , n = 6 ） ,99. 01% （RSD = 0.47 % , n = 6 ）and 99.00% （RSD=0.82%, n = 6 ） , respectively. Conclusion： The method is simple, accurate, reliable and can be applied to the quality control of Tuire Jiedu Injection .