HPLC法测定银花清咽含片中绿原酸木犀草苷连翘苷含量

目的建立HPLC法测定银花清咽含片中绿原酸、木犀草苷和连翘苷含量的方法。方法 Venusil MP C18柱(4.6 mm×250 mm,5μm),乙腈-0.4%磷酸(10∶90)为流动相,流速:1 ml.min-1,检测波长327 nm,柱温:25℃,检测绿原酸;采用Agilent ZORBAX SB-phenyl柱(4.6 mm×250 mm,5μm),乙腈为流动相A,0.5%冰醋酸为流动相B梯度洗脱,梯度程序为0min(A:10%,B:90%)→15 min(A:18%,B:82%)→30 min(A:18%,B:82%)→40 min(A:30%,B:70%)→45 min(A:40%,B:60%),流速:1 ml.min-1,检测波长:350 nm,柱温:30℃,检测木犀草苷;采用乙腈-水(24∶76)为流动相,流速:1 ml.min-1,检测波长:277 nm,柱温:30℃,检测连翘苷。结果绿原酸、木犀草苷、连翘苷分别在0.3～2.4、0.012～0.096、0.26～2.08μg范围内线性关系良好,平均回收率分别为99.78%、99.90%、100.09%,RSD分别为0.54%、0.46%、0.21%(n=6)。结论该方法重复性好、准确、专属性强,可用于银花清咽片中绿原酸、木犀草苷、连翘苷的含量测定,为较全面地控制银花清咽片的质量提供简便可靠的方法。

Determination of Chlorogenic Acid Galuteolin and Forsythin in Yinhuaqingyan Tablets by HPLC

Objective To establish an HPLC method for determination of chlorogenic acid, galuteolin and for- sythin in Yinhuaqingyan tablets. Methods Chlorogenic acid was determined by HPLC with Venusil MP C18 column （4.6 mm ×250 mm,5 μm）. The mobile phrase was composed of acetonitrile-0.4% phosphoric acid （ 10： 90）. The de- tection wavelength was 327 nm and the column temperature was 25 ℃. Galuteolin in honeysuckle was determined on an Agilent ZORBAX SB-phenyl column（4.6 mm× 250 ram, 5 μm）with acetonitrile as mobile phrase A and 0.5% glacial acetic acid as mobile phrase B. The gradient procedure was 0 min（A：10% ,B：90% ）→15 min（A：18% ,B： 82% ） →30 min （ A ： 18%, B ：82% ） →40 min （ A： 30%, B ：70% ） →45 min （ A ：40%, B ：60% ）. The detection wave- length was set at 350 nm with a temperature of 30 ℃. Forsythin was eluted by acetonitrile-water （24： 76） and detec- ted at 277 nm when the column temperature was 30 ℃. Results The linear range of chlorogenic acid, galuteolin and forsythin was 0.3-2.4,0.012-0.096 and 0.26-2.08 p,g,respectively. The average recovery was 99.78% ,99.90% and 100.09% with an RSD of 0.54% ,0.46% and 0.21%. Conclusion This method is simple, accurate, specific and can be used for determination of chlorogenic acid, galuteolin and forsythin in Yinhuaqingyan tablets.